skip to main content
Show Results with:

DIABETES EXPERIMENTAL

Chan, John S. d. et al.

Nephrology dialysis transplantation volume 29 issue S3 page iii168 -- Oxford University Press

Online access

  • Title:
    DIABETES EXPERIMENTAL
  • Author: Chan, John S. d.;
    Pontrelli, P.;
    Conserva, F.;
    Papale, M.;
    Accetturo, M.;
    Gigante, M.;
    Vocino, G.;
    Dipalma, A.M.;
    Grandaliano, G.;
    Di Paolo, S.;
    Gesualdo, L.;
    Abdo, Shaaban;
    Ghosh, Anindya;
    Alquier, Thierry;
    Chenier, Isabelle;
    Filep, Janos G.;
    Ingelfinger, Julie R.;
    Zhang, Shao-Ling;
    Ross, Edward A;
    Willenberg, Bradley J;
    Oca-Cossio, Jose;
    Clapp, William L;
    Terada, Naohiro;
    Abrahamson, Dale R;
    Ellison, Gary W;
    Matthews, Clayton E;
    Batich, Christopher D;
    Ihoriya, Chieko;
    Satoh, Minoru;
    Sasaki, Tamaki;
    Kashihara, Naoki;
    Piwkowska, Agnieszka;
    Rogacka, Dorota;
    Angielski, Stefan;
    Jankowski, Maciej;
    Franzén, Stephanie;
    Pihl, Liselotte;
    Khan, Nadeem;
    Gustafsson, Håkan;
    Palm, Fredrik;
    Koszegi, Sandor;
    Hodrea, Judit;
    Lenart, Lilla;
    Hosszu, Adam;
    Wagner, Laszlo;
    Vannay, Adam;
    Tulassay, Tivadar;
    Szabo, Attila;
    Fekete, Andrea;
    Aoki, Rieko;
    Sekine, Fujio;
    Kikuchi, Kaori;
    Miyazaki, Shigeaki;
    Yamashita, Yusuke;
    Itoh, Yoshiharu;
    Kölling, Malte;
    Park, Joon-Keun;
    Haller, Hermann;
    Thum, Thomas;
    Lorenzen, Johan;
    Hirayama, Aki;
    Yoh, Keigyou;
    Ueda, Atsushi;
    Itoh, Hiromu;
    Owada, Shigeru;
    Kokeny, Gabor;
    Szabó, Lilla;
    Fazekas, Krisztina;
    Rosivall, László;
    Mózes, Miklós M;
    Kim, Yaeni;
    Koh, Eun Sil;
    Lim, Ji Hee;
    Kim, Min Young;
    Chang, Yoon Sik;
    Park, Cheol Whee;
    Kim, Hyung Wook;
    Shin, Byung Chul;
    Kim, Hyun Lee;
    Chung, Jong Hoon;
    Chan, Jenq-Shyong;
    Wu, Tao-Cheng;
    Chen, Jaw-Wen;
    Clotet, Sergi;
    Soler, María José;
    Rebull, Marta;
    Pascual, Julio;
    Riera, Marta;
    Patinha, Daniela;
    Afonso, Joana;
    Sousa, Teresa;
    Morato, Manuela;
    Albino-Teixeira, Antonio;
    Kim, Hyunwook;
    Min, Hye Sook;
    Kang, Min-Jung;
    Kim, Jung Eun;
    Lee, Ji-Eun;
    Kang, Young Sun;
    Cha, Dae Ryong;
    Jo, Young-Il;
    Seo, Eun-Hye;
    Kim, Joo-Dong;
    Lee, Seung-Hyun;
    Jorge, Luciana;
    Silva, Kleiton A.S.;
    Luiz, Rafael S.;
    Rampaso, Rodolfo R.;
    Lima, Weslei;
    Cunha, Tatiana S.;
    Schor, Nestor;
    Lee, Hong Joo;
    Park, Ji Yun;
    Kim, Seul Ki;
    Moon, Ju Young;
    Lee, Sang Ho;
    Ihm, Chun Gyoo;
    Lee, Tae Won;
    Jeong, Kyung Hwan;
    Moon, Ju-Young;
    Kim, Seulki;
    Park, Ji-Yun;
    Kim, Se-Yun;
    Kim, Yang-Gyun;
    Jeong, Kyung-Hwan;
    Lee, Sang-Ho;
    Ihm, Chun-Gyoo;
    Marques, Catarina;
    Mega, Cristina;
    Gonçalves, Andreia;
    Rodrigues-Santos, Paulo;
    Teixeira-Lemos, Edite;
    Teixeira, Frederico;
    Fontes Ribeiro, Carlos;
    Reis, Flávio;
    Fernandes, Rosa;
    Sutariya, Brijesh Kantilal;
    Badgujar, Lohit Bhagwan;
    Kshtriya, Ashish Arun;
    Saraf, Madhusudan Natvarlal;
    Chiu, Chien-Hua;
    Lee, Wen-Chin;
    Chau, You-Ying;
    Lee, Lung-Chih;
    Lee, Chien-Te;
    Chen, Jin-Bor;
    Dahan, Inbal;
    Nakhoul, Farid;
    Thawho, Nadia;
    Ben-Itzhaq, Ofer;
    Levy, Andy P;
    Conserva, Francesca;
    Pontrelli, Paola;
    Accetturo, Matteo;
    Cordisco, Giorgia;
    Fiorentino, Loredana;
    Federici, Massimo;
    Grandaliano, Giuseppe;
    Di Paolo, Salvatore;
    Gesualdo, Loreto;
    Wystrychowski, Grzegorz;
    Havel, Peter J.;
    Graham, James L.;
    Żukowska-Szczechowska, Ewa;
    Obuchowicz, Ewa;
    Psurek, Anna;
    Grzeszczak, Władysław;
    Wystrychowski, Antoni;
    Gimeno, Javier;
    Almeida, Beatriz Zocoler de;
    Seraphim, Deborah Chianelli Costalonga;
    Punaro, Giovana;
    Nascimento, Marcos;
    Mouro, Margaret;
    Lanzoni, Valéria Pereira;
    Lopes, Guiomar Silva;
    Higa, Elisa Mieko Suemitsu;
    Roca-Ho, Heleia;
    Márquez, Eva;
    Soler, Mª José
  • Found In: Nephrology dialysis transplantation volume 29 issue S3 page iii168
  • Subjects: Nephrology; Hemodialysis; Kidneys; Transplantation; Hemodialysis; Kidneys; Transplantation; Nephrology
  • Rights: LegalDeposit; Reading Room Access
  • Publication Details: Oxford University Press
  • Abstract: Introduction and Aims: Angiotensinogen (Agt) is the sole precursor of all angiotensins. We have previously reported that high glucose stimulates renal Agt gene expression via nuclear factor erythroid 2-related factor 2 (Nrf2) in type 1 diabetic Akita mice. The present studies hypothesized that insulin inhibits renal angiotensinogen (Agt) gene expression via down-regulation of Nrf2 and thus prevents systemic hypertension in type 1 diabetic Akita mice and in wild type (WT) mice infused with insulin and various concentration of D-glucose to maintain normal blood glucose levels. Methods: Male Akita mice were studied from 10-16 weeks of age with or without insulin implants (Linβit) from week 12. Age- and sex-matched non-Akita littermates served as wild type (WT) controls. We also did insulin clamp studies in WT mice (age: 8-10 weeks) using insulin infusion (~3 ng/min/kg) plus variable concentrations of D-glucose to maintain euglycemia for 3 hours (hyperinsulinemia-euglycemic clamp). WT mice infused with saline served as controls. Kidneys were analyzed for Nrf2, Agt, and heterogeneous nuclear ribonucleoprotein F and K (hnRNP F/K) expression by immunohistochemistry. mRNA and protein expression in renal proximal tubules (RPTs) were evaluated by real time-qPCR and Western blotting, respectively. Immortalized rat renal proximal tubular cells (IRPTCs), stably transfected with plasmid pGL4.20 and containing either rat Nrf2, Agt, hnRNP F or hnRNP K gene promoter, were also studied in vitro. Results: In Akita mice, insulin treatment normalized blood glucose levels, reversed systemic hypertension and renal oxidative stress, inhibited renal Nrf2 and Agt gene expression, whereas it enhanced hnRNP F/K gene expression and attenuated renal hypertrophy, glomerular hyperfiltration and tubulointerstitial fibrosis. In WT mice subjected to euglycemic insulin clamp, renal Agt and Nrf2 gene expression was down-regulated, whereas hnRNP F/K gene expression was up-regulated as compared to saline-infused control WT mice. In vitro, insulin inhibited Nrf2 and Agt gene promoter activity, and stimulated hnRNP F/K gene promoter activity in high glucose media via the p44/42 mitogen-activated protein kinase signaling pathway. Transfection with Nrf2 small interfering RNA enhanced insulin inhibition of Agt and stimulated hnRNP F/K gene promoter activity in IRPTCs. Conclusions: Our data indicate that insulin prevents hypertension and attenuates kidney injury, at least in part, through suppressing renal Nrf2 and Agt gene transcription and up-regulating hnRNP F/K gene expression in diabetic Akita mice, independent of its glucose lowering effect.Introduction and Aims: We have studied tissue regeneration by seeding decellularized kidneys with progenitor cells, and our previous results have shown endothelial differentiation of mouse embryonic stem cells in the vasculature of rat scaffolds. In that a barrier to pancreatic islet cell transplantation is lack of adequate blood supply, we hypothesize that regenerated renal scaffold vessels (glomerular and peritubular capillaries) can serve to vascularize insulin-secreting cells seeded into what was the collecting system (Bowman’s space and tubules, respectively). For the ultimate goal of human xenotransplantation, this “trans-organ” would have a pig kidney scaffold seeded by endothelial and islet cell lineages grown from induced pluripotent stem cells (iPSCs) derived from the diabetic patient. Methods: Based on our murine findings, we tested new protocols to decellularize, sterilize, and seed kidneys harvested from 12 kg juvenile pigs, using Triton X-100, SDS, DNase, and peracetic acid. Integrity of the vascular and collecting systems was studied using light microscopy and after infusion of fluorescent 10 micron polystyrene microspheres. Cell seeding techniques were developed using positive and/or vacuum pressure. Fluorescent cells were injected anteriorly (immortalized mouse lung fibroblasts or HeLa) via the artery and retrograde (immortal islet cells: TC-Tet or BLox5) via the ureter. Histology was performed to assess cell delivery patterns, integrity of the vascular and collecting system compartments, and for insulin secretion. Results: Microscopy demonstrated preservation of the decellularized renal scaffold architecture. Microspheres were visualized antegrade as far as the glomerular tufts, and retrograde into Bowman’s space without mixing between compartments. Vacuum-assisted seeding improved cell delivery into vasculature and collecting systems: For the former, by demonstrating appropriate cells in the glomerular and peritubular capillary vasculature (e.g. GFP+). For the latter, by showing pancreatic islet cells in Bowman’s space and tubules (e.g. insulin+). Conclusions: We have successfully developed protocols to transition from our rat model to pig kidneys for decellularization, sterilization and cell seeding. We selectively deployed cells antegrade through the vasculature into glomerular tufts, and retrograde up tubules into Bowman’s space. Pancreatic islet cells successfully grew in the collecting system and demonstrated insulin production. This approach is very promising for tissue engineering a pig kidney trans-organ using a patient’s iPSCs, and thereby crafting a vascularized pancreas xenotransplant.Introduction and Aims: The nuclear factor erythroid 2-like factor 2 (Nrf2) is an important oxidative stress-responsive transcription factor with a vital role in combating oxidative damage. Nrf2 plays a critical part in basal activity and coordinated induction of genes encoding numerous antioxidant and phase II detoxifying enzymes and related proteins. In many clinical trials, HMG-CoA reductase inhibitors (statins) have shown clear benefits in cardiovascular disease beyond their lipid-lowering actions, as they also function as antioxidants. Statins not only decrease cellular reactive oxygen species production but also enhance the antioxidant response by upregulating the expression of many antioxidants. We hypothesized that statin reduces oxidative stress through the Nrf2 signaling pathway and has renoprotective effects against diabetic nephropathy. The aims of this study were to characterize the effect of rosuvastatin on albuminuria in Akita diabetic mice and to identify the molecular mechanisms of Nrf2 activation by rosuvastatin. Methods: Wild type (WT) and Akita diabetic mice (AKITA) were treated with Rosuvastatin (RSV) for 4 weeks. Urinary albumin excretion and renal histology were examined. Nrf2-antioxidant response element (ARE) activity was measured in human umbilical vein endothelial cell (HUVEC) with luciferase assay after transfection of reporter plasmids containing AREs. The expression of Nrf2-regulated genes was also examined. Results: Increased urinary albumin excretion in AKITA mice was significantly reduced by RSV treatment. The amount of lectin-stained glomerular endothelial surface layer, important for permselectivity in the vascular wall, was significantly reduced in AKITA mice and preserved with RSV treatment. RSV significantly increased the transcriptional activity of the AREs and subsequent expression of Nrf2-regulated genes in HUVEC. Additional experiments with cycloheximide and actinomycin D indicated that RSV extended the half-life of Nrf2 protein. Furthermore, RSV increased p21cip1 expression and thereby inhibited degradation of Nrf2 through direct binding of Nrf2 with p21cip1. Conclusions: We demonstrated that RSV activates transcription factor Nrf2 in endothelial cells. RSV activated Nrf2 dependent anti-oxidant genes and increased anti-oxidant capacity, which may reduce albuminuria through the preservation of endothelial cells in Akita diabetic mice. Oxidative stress is the basis of the morbidity of diabetic nephropathy including chronic kidney disease and statins treatment is a useful therapeutic strategy to enhance anti-oxidative capacity.Introduction and Aims: Podocytes are uniquely sensitive to insulin, they have demonstrated similarities to skeletal muscle and fat cells with respect to insulin stimulated glucose uptake kinetics and the expression of glucose transporters (GLUT). Insulin signaling is regulated by oxidative stress and intracellular energy levels. We demonstrated recently that superoxide anion generation or insulin increases dimerization of protein kinase G type I alpha (PKGIalpha) subunits, leading to podocyte dysfunction. Here we investigated whether PKGI-dependent pathway can modulate the insulin signaling and glucose transport system in rat cultured podocytes. Methods: To determine whether cGMP-dependent protein kinase is involved in the insulin regulation of glucose transport, we measured insulin-dependent glucose uptake into cultured rat podocytes under conditions of modified PKG activity using pharmacological (PKG activator or inhibitor) and biochemical (siRNA PKGIalpha, siRNA insulin receptor) means. Protein expression was measured with Western blot and immunofluorescence. Glucose uptake was measured with radioactive method. Results: Podocytes were insulin-sensitive, and insulin (300nM, 3 min) stimulation increased glucose uptake by about 50%. Hydrogen peroxide or 8-Br-cGMP (100µM, 3 min, PKG activators) significanly increases glucose uptake to podocytes. To determine the role of PKGI on insulin-induced glucose uptake, we evaluated the effect of Rp-8-cGMPS (100µM, 20 min preincubation, PKGI inhibitor). We found that Rp-8-cGMPS attenuated insulin- and hydrogen peroxide-induced stimulation of glucose uptake. The similar effect we observed after knock down PKGIα or IR protein expression with small-interfering RNA. The effect of insulin on IR phosphorylation was also significantly reduced by about 25% in the presence of Rp-8-cGMPS. Moreover, the effect of insulin on Akt phosphorylation was abolished in the presence of PKG inhibitor. The immunofluorescence experiments showed that PKGI activators likewise insulin, caused substantial changes in the subcellular localization of the GLUT4 transporter in podocytes. The intensity of GLUT4 immunostaining increased close to the cell surface. Preincubation podocytes with PKGI inhibitor abolished this effect. Conclusions: The results of the present study indicate that insulin activated the insulin signaling pathway, glucose transport and GLUT4 translocation via protein kinase G type I in cultured rat podocytes. This signaling may play a potential role in the prevention of insulin resistance under conditions associated with oxidative stress.Introduction and Aims: Chronic hyperglycemia, a characteristic feature of type 2 diabetes, causes renal lesions in 20-30% of patients leading to DN, the principal cause of end-stage renal disease. Tubular cells play an important role in the pathogenesis of DN, but the molecular damage induced by hyperglycemia is almost unclear. Among the pathways known to be involved in the onset of DN, the ubiquitin pathway is emerging as a possible key-player. UBE2V1, an ubiquitin-conjugating E2 enzyme variant, mediates the formation of lysine 63 (lys63)-linked ubiquitin chains, affecting protein localization and cell signaling. Aim of our study was to evaluate the role of protein ubiquitination in response to hyperglycemia and its possible involvement in the progression of tubular damage in DN. Methods: Human tubular cells (HK2) were grown in normal (5.5 mM) and high (30 mM) glucose (HG) conditions. Glucose effects on UBE2V1 expression were evaluated by real-time PCR and immunoblotting on cell extracts, and immunohistochemistry on formalin-fixed paraffin-embedded (FFPE) biopsies of six DN patients and six normal controls. Protein lysates of HK2 cells were immunoprecipitated to identify lysine 63-ubiquitinated proteins by MALDI-TOF/MS-MS. Lysine63-Ubiquitination of the identified proteins was confirmed by western blotting and confocal microscopy Results: UBE2V1 gene expression was increased in HK2 cells after HG stimulation (1.51 fold). Also protein expression of UBE2V1 and Lys63-ubiquitinated proteins was increased on HK2 cells under HG conditions in vitro, and in vivo on FFPE tissues biopsies of patients with DN (UBE2V1, p=0.004; Lys63: p= 0.008 DN vs control), with a significant increased expression at tubular level. To identify which proteins underwent a specific Lysin63-ubiquitination under HG conditions, HK2 protein lysates were immunoprecipitated with an anti-ubiquitin antibody and blotted with an antibody against lys63-linked ubiquitin chains. The identification of lys63-linked ubiquitinated proteins by MALDI-TOF/MS-MS led to the discovery of 28 proteins specifically ubiquitinated in Lys63 under HG conditions, mainly involved in cellular assembly and organization (IPA score=50). Among the proteins identified, beta actin lys63 ubiquitination was confirmed by western blotting under chronic HG exposure (p=0.05), with a subsequent depolymerization of actin cytoskeleton (confocal microscopy), and the loss of vimentin (flow cytometry). UBE2V1 silencing in HK2 cells under HG conditions reported to basal condition the actin ubiquitination in Lys63 (western blotting). Conclusions: These findings suggest that UBE2V1 and Lys63-poly-ubiquitinated proteins increase under HG conditions and persist in DN patients, mainly affecting cytoskeleton organization. This event could compromise cell survival and function, leading to progression of tubular damage during DN. Moreover, cytoskeletal ubiquitination could represent a potential therapeutic target to reduce tubular damage in the progression of DN.Introduction and Aims: Tissue hypoxia has been postulated as a unifying mechanism for the onset and progression of kidney disease. However, renal hypoxia has to be present before any detectible signs of nephropathy, which has been technically difficult. Methods: In this study, we utilized implantable crystals of lithium phthalocyanine (LiPc) possessing an oxygen-dependent electron paramagnetic resonance (EPR) signal. Thus, absolute oxygen tensions can be monitored repetitively in mice using L-band EPR. After implantation of the LiPc crystals into kidney cortex of adult male NMRI mice, seven repetitive measurements of oxygen tension were performed at regular intervals starting before induction of diabetes (alloxan 75 mg/kg i.v.) and ending 15 days thereafter. All results were compared to a separate normoglycemic control group. Results: Baseline cortical oxygen tension was similar in the two groups (51±2 and 49±2 mmHg; ns). Whereas cortical oxygen tension was unaltered in the normoglycemic control group, oxygen tension decreased already three days after the onset of hyperglycemia in the diabetic group (-16±3 mmHg; P<0.001 vs. baseline, -18±5 mmHg; P<0.01 vs. controls) and remained lower throughout the study period. Conclusions: In conclusion, these results demonstrate that kidney cortex hypoxia occurs very early after the onset of diabetes. Thus, intrarenal hypoxia occurs before the later development of diabetic nephropathy, which is a prerequisite for hypoxia being a contributing mechanism.Introduction and Aims: Diabetic nephropathy (DN) is the leading cause of chronic renal failure in adults, however its treatment is unsolved and the exact pathomechanism is yet unknown. In diabetes the the increased renin-angiotensin-aldosterone system (RAAS) activity contributes to the development of renal fibrosis. Platelet-derived growth factor (PDGF) is one of the key molecules of fibrosis, but its signal transduction pathway DN is still not clarified. Methods: : Five weeks after streptozotocin (65 mg/bwkg, i.p.) induced diabetes male Wistar rats were treated for two weeks per os with enalapril (40 mg/bwkg/day), ramipril (10 mg/bwkg/day), losartan (20 mg/bwkg/day), spironolactone or eplerenone (50 mg/bwkg/day). Vehicle-treated healthy or diabetic animals served as controls (n=6/group). Mesangial matrix expansion and renal fibrosis were evaluated on PAS, Masson and fibronectin stained sections. PDGF and αSMA protein level were measured in the kidney. Human proximal tubular cells (HK2) were cultured in normal (5.5 mM) or high (35 mM) glucose medium. Isosmotic controls were cultured in a mannitol (35 mM). First, we measured the glucose-induced PDGF production of HK2 cells. Then to verify the effect of PDGF, normal rat kidney (NRK-49F) fibroblasts were treated with PDGF (10 ng/mL) and αSMA protein level was detected. Results: Impaired renal function and metabolic parameters confirmed the development of DN. The decline in GFR was ameliorated by RAAS blockers. Increased mesangial matrix expansion and interstitial fibrosis in DN were reduced by all RAAS inhibitors, while the amount of fibronectin was only decreased by ACE inhibitors and eplerenone. In high glucose PDGF production of tubular cells was increased, but not in mannitol. PDGF- treated fibroblasts produced more αSMA. Diabetes induced elevation of renal PDGF and αSMA protein were decreased by RAAS blockers. Conclusions: High glucose increases the PDGF production of proximal tubular cells, that in turn induces the αSMA production in renal fibroblast. , This mechanism also contributes to the development of renal fibrosis seen in DNP. RAAS blockers ameliorates this process by directly acting on renal fibroblasts which could serve as a new therapeutic potential in the treatment of renal fibrosis. Grants:LP008/2011, OTKA-NK84087/2010,K100909, -K108688, KMR12-1-2012-0074Introduction and Aims: Metabolic syndrome is known to be an important risk factor involved in the development of diabetic nephropathy.An oral adsorbent AST-120 has been used clinically as a drug for treatment of chronic kidney disease (CKD) patients to slow the progression of CKD.However, there is little evidence when AST-120 should be prescribed for subjects with early stage overt diabetic nephropathy.In this study, we investigated whether AST-120 has an effect on the early stage of nephropathy using SHR/NDmc-cp (SHR/ND), a rat model of metabolic syndrome/ type 2 diabetes. Methods: Male SHR/NDmc-cp (SHR/ND) rats, aged 7 weeks, were administered 0% or 8% AST-120 for 12 weeks in their diets. WKY rats were used as a normal. At every 4 weeks, serum and 24-hour urine samples were collected for biomedical studies. We analyzed serum metabolites in normal and SHR/ND rats with or without AST-120 for 8 weeks by capillary electrophoresis mass spectrometry with time-of-flight (CE-TOFMS) and applied CE-TOFMS data to principal component analysis (PCA).We also examined the urinary 8-OHdG excretion by ELISA as an oxidative stress maker and the podocyte foot process width (FPW) as its injury by Transmission Electron Microscopy. Results: AST-120-administered SHR/ND rats showed significantly lower levels of urinary protein excretion, urinary albumin excretion, urinary 8-OHdG excretion and the FPW as compared with SHR/ND rats. The FPW was significantly correlated with the levels of urinary protein excretion (r = 0.9498) and urinary albumin excretion (r = 0.9532). PCA score plot showed clear separation among three groups (Normal, SHR/ND and AST-120-administered SHR/ND). We could detect 40 metabolites, such as o-hydroxybenzoic acid, hippuric acid and indole-3-acetic acid, which accumulated in the serum of SHR/ND rats, and of which serum levels were reduced by administration of AST-120. Conclusions: The amelioration of podocyte injury by AST-120 may contribute to the reduction of proteinuria and albuminuriaIt indicates that the administration of AST-120 at an early stage of diabetic nephropathy has a protective effect on the disease progression.Introduction and Aims: The long-term complications arising from diabetes become nowadays an increasing challenge. Pathological changes in the kidney are crucially involved in the detrimental effects on the cardiovascular system. Hyperglycaemia, mainly present in the diabetic state, results in the activation of PKC-β and its downstream pathways finally lead to diabetic nephropathy. Methods: Using a diabetic streptozotocin mouse model, we analysed the effects of diabetes in wild type and PKC-β knock out mice. Mesangial cells were isolated for further analysis. To reveal relevant deregulations of microRNAs, we performed a microRNA microarray. MicroRNAs are small noncoding nucleotides regulating gene expression, such as those involved in fibrosis development. In vitro, mesangial cells were subjected to hyperglycaemia as well as TGF-β treatment. The expressions of fibrosis-associated microRNAs and fibrotic genes were determined by real time PCR. Involved signalling pathways were investigated by western blotting. PKC-β was pharmacologically inhibited by treating mesangial cells with the specific PKC inhibitor Ruboxistaurin. Transcriptional regulation was assessed by Chromatin immunoprecipitation and electrophoretic mobility shift assay (EMSA) analysis. Results: Fibrosis-associated microRNAs were upregulated in the streptozotocin induced diabetic wild type mouse model compared to the streptozotocin induced diabetic PKC-β knock out mouse model. In vitro, the treatment with TGF-β for 24h led to a significantly increased expression of various fibrosis-associated mature as well as primary microRNAs (all p<0.05), indicating transcriptional activation. In addition, fibrotic genes were upregulated by TGF-β treatment (Col1a2, p=0.004; CTGF, p=0.001; Fibronectin, p=0.04). On the protein level, the amounts of both phosphorylated AKT and phosphorylated ERK were significantly increased (both p<0.05) after TGF-β subjection. Impressively, the pharmacological inhibition of PKC-β resulted in a reduction of fibrosis-associated microRNAs. This was associated with normalization of CTGF expression (p=0.05). Finally, analysis of the transcription factor AP-1 by EMSA revealed an enhanced signal for AP-1 in the nuclear extract obtained from mesangial cells treated with TGF-β for 24h compared to the control treatment. Conclusions: These results indicate that PKC-β plays a pivotal role in regulating diabetes-induced renal endpoints as well as fibrosis-associated microRNAs. Influencing this pathway may represent a prospective therapeutic approach to avert diabetic nephropathy and its resulting complications.Introduction and Aims: Previously we have demonstrated that oxidative and nitrosative stress played a critical role in renal injury of streptozotocin-induced diabetic Nrf2-deficient (Nrf2 KO) mice. In this study we examined whether TCV-116 (candesartan cilexetil), an Angiotensin-II-receptor blocker, ameliorates this damaging process. Methods: 8 weeks-old Nrf2 KO and wild type mice were induced diabetes by a single intraperitoneal injection of streptozotocin (STZ). TCV-116 was given in drinking water at the dose of 5 mg/kg/day for 24 weeks from 2 weeks after diabetes development. Oxidative stress was evaluated by measuring scavenging activities against hydroxyl radical and superoxide using a newly-developed electron paramagnetic resonance (EPR)-based spin trapping method. Results: Injection of STZ equally increased plasma glucose level in both wild type and Nrf2 KO mice, and TC116 made no difference on it. The survival rate of diabetic Nrf2 KO (dKO) mice was shorter than that of non-treated wild type (cWild), diabetic wild type (dWild) and non-treated Nrf2 KO (cKO) mice. TCV-116 significantly improved the survival rate of dKO mice. TCV-116 treatment also improved serum creatinine level of dKO mice at 10 weeks, but this effect was not significant at 24 weeks. Histologically, induction of diabetes induced glomerular hypertrophy and mesangial expansion both in dWild and dKO mice, and TCV-116 treatment improved the lesions in a same manner. According to nitrosative stress, urinary NOx excretions were significantly increased both in Nrf2 KO and wild type mice. TCV-116 significantly decreased urinary NOx at 10 weeks, but not significant at 24 weeks. TCV-116 also improved serum hydroxyl radical scavenging activity and serum glucose-derived AGEs (Glc-AGEs) levels at 24 weeks. No effects on superoxide scavenging activity were confirmed. Conclusions: These findings suggest that TCV-116 treatment improved survival rate and ameliorate renal failure due to reduced oxidative and nitrosative stresses in diabetic Nrf2 KO mice.Introduction and Aims: Diabetic nephropathy is associated with functional imbalance of matrix metalloproteinases (MMPs) and their inhibitors (TIMPs), and renal TIMP-1 overexpression has been reported in experimental and human studies. EGR1 is an early transcription factor which regulates collagen synthesis and modulates fibroblast proliferation. However, its association with diabetes or TIMP-1 has not been clarified yet. We investigated the progression of nephropathy in type-1 diabetic TIMP-1 knockout (KO) mice and its association with the profibrotic factor EGR1. Methods: Diabetes was induced in 6 week-old male TIMP-1 KO (TIMP-1, n=8) and C57Bl6 wild type (WT, n=5) mice weighing 20-22 grams with daily intraperitoneal streptozotocin (50 mg/kg/day) injections for 5 consecutive days. Fasting blood glucose was monitored once weekly. After 8 weeks, kidneys were analyzed for histology and mRNA expression levels. Results: Blood glucose similarly exceeded 30 mmol/l in both groups. TIMP-1 mice presented 52% lower urinary protein/creatinine ratio (WT: 49±9 vs TIMP-1: 23±8 mg/mg, p=0.0001) and 21% lower serum urea levels (WT: 53±9 vs TIMP-1: 42±6 umol/l, p=0.03), Reduced glomerulosclerosis scores (WT: 1.8±0.5 vs TIMP-1: 1.0±0.2, p=0.037) and tubular damage scores (WT: 1.2±0.3 vs TIMP-1: 0.8±0.2, p=0.018) were accompanied by 42% reduction in EGR-1 mRNA expression (WT: 0.9±0.2 vs TIMP-1: 0.6±0.1 relative expression, p=0.03) and by 60% decrease in the number of EGR1 positive tubular cells in TIMP-1 mice (WT: 120±50 vs TIMP-1: 23±9 cells/field, p=0.014). Conclusions: Our results suggest that TIMP-1 contributes to diabetic nephropathy by direct inhibition of MMPs and by its effect on the transcription factor EGR1.Introduction and Aims: Purple corn anthocyanins have been reported to possess antidiabetic, antiangiogenic, and anticarciogenic activities. However, little studies have been investigated the effects of anthocyanins on diabetic nephropathy. The activations of AMP-activated protein kinase (AMPK), silent information regulator T1 (SIRT1) and peroxisome proliferator-activated receptor (PPAR)γ co-activator1α (PGC-1α) play key roles in the regulation of lipid and glucose homeostasis, and in the control of oxidative stress. We investigated whether anthocyanins may have protective effects on the kidney in type 2 diabetes and human glomerular endothelial cells (HGECs). Methods: Male C57 BLKS db/db mice and db/m controls at 8 weeks of age were divided to receive either a regular diet chow or a diet containing anthocyanin (10 mg/kg/day, n=8, respectively). Mice were followed for 12 weeks and were evaluated about renal functional and pathologic phenotypes and the AMPK- PPARα-PGC-1α-ERR-1α pathway. Results: Anthocyanin treatment ameliorated albuminuria (P<0.001) in db/db mice with the same degree of blood glucose and HbA1c concentraions. The mesangial area expansion, inflammatory cell infiltration, and the accumulation of intra-renal free fatty acid and triglycerides were observed in db/db mice. A downregulation of AMPK suppressed PPARα-PGC-1α and ERR-1α expressions in the kidney, which led to oxidative stress and decreases fatty acid oxidation. Treatment of anthocyanin increased the AMPK expression and subsequently activated the PPARα-PGC-1α-ERR-1α and lipid accumulation in renal cortex. In cultured HGECs, suppressed AMPK-PPARα-PGC-1α-ERR-1α signaling in high-glucose media reversed by anthocyanin, which was associated with decreases in oxidative stress and apoptosis. Conclusions: In conclusion, the results suggest that natural anthocyanin product improves lipotoxicity through activation of the AMPK- PPARα-PGC-1α-ERR-1α signaling and may be a potentially therapeutic modality for type 2 diabetic nephropathy.Introduction and Aims: The calcimimetic, cinacacet, which activates Ca-sensing receptor (CaR), is used to treat hyperparathyroidism. Interestingly, the CaR also modulates ion-channel/transporter activity, peptide secretion, cell proliferation and apoptosis, chemotaxis, and oncogene modulation. In cardiovascular system, the CaR is also present on endothelial cells and simulated the production of NO. A decrement of in NO bioavailability and increased the generation of potentially toxic reactive oxygen species are critical to the pathogenesis of diabetic vascular complication, including diabetic nephropathy. Therefore, we evaluated the renoprotective effect, especially endothelial dysfunction, of cinacalcet agonist glucotoxicity through the change of oxidative stress and eNOS-NO pathway on diabetic nephropathy in db/db mice and human glomerular endothelial cells (HGECs). Methods: Male C57 BLKS db/db mice and db/m controls at 8 weeks of age were divided to receive either a regular diet chow or a diet containing cinacalcet (10 mg/kg; 0.001% wt/wt, n=8, respectively). Mice were followed for 12 weeks and were evaluated about renal functional and pathologic phenotypes and eNOS-NO pathway. Results: Cinacalcet ameliorated albuminuria and decreased urine volume in db/db mice without the change of blood glucose concentrations. The mesangial area expansion, inflammatory cell infiltration and the number of apoptotic endothelial cells in the glomerulus were observed in db/db mice, which were all restored by cinacalcet treatment. Treatment of cinacalcet increased the level of phospho-Ser1177 eNOS-NO and subsequently increased the ratio of Bcl-2/Bax in renal cortex. Urinary 8-OH-deoxyguanosin concentrations were decreased by cinacalcet treatment along with increases in SOD1 and SOD2 expression. In cultured HGECs, suppressed phospho-Ser1177 eNOS-NO and Bcl-2/Bax expressions were reversed by cinacalcet treatment, which was associated with decreases in oxidative stress and apoptosis. Conclusions: In conclusion, the results suggest that calcimimetic cinacalcet improves glucotoxicity through activation of the eNOS-NO signaling in the kidney and may be a potentially therapeutic modality for type 2 diabetic nephropathy.Introduction and Aims: Diabetic nephropathy is one of the most serious complications in diabetes mellitus and has been the most common cause of end-stage renal disease. Green tea extracts have antioxidant properties, and (-)-epigallocatechin 3-O-gallate (EGCG) is known to be the most abundant in green tea. The present study evaluates effects of EGCG against streptozotocin (STZ)-induced diabetic nephropathy in mice. Methods: The mice (n=50) were divided into 5 groups. Control group (n=10) was intraperitoneal (IP) injected 0.9% saline, Streptozotocin (STZ) group (n=10) was IP injected STZ 200mg/Kg and induced diabetic nephropathy. At 16 weeks, EGCG groups (n=10/each 3 group) were received EGCG 50 mg/kg (EGCG50 group), 100 mg/kg (EGCG 100mg) and 200 mg/kg (EGCG200 group) body weight by subcutaneous injection for 7days. Serum glucose, blood urea nitrogen, serum creatinine, urine volume and urine protein amounts were measured. Western blot assay of osteopontin (OPN) was compared for the different groups. Histopathologic examination and immunohistochemical staining of mice kidney were performed. Results: Compared with control group, STZ-group showed an increase in blood glucose, blood urea nitrogen, creatinine levels and urine protein amounts, and a decrease in body weight. All the above parameters were significantly reversed with EGCG treatment, especially EGCG100 group. After STZ injection, there were a diabetic glomerulosclerosis with increased renal OPN accumulation and its protein expression in the kidney cortex. EGCG-treated mice kidney showed a reduced expression of above parameters and a reserved pathologic findings. Conclusions: These results indicated that EGCG 100 mg/kg had effective protection against STZ-induced diabetic nephropathy in mice by OPN suppression. The potential use of EGCG in the treatment of diabetic nephropathy should be further explored.Introduction and Aims: Microvascular dysfunction is an integral component of the pathological complications that occur in the diabetic foot. In addition to interventional revascularization, the most important alternative approach to increase blood flow to the ischemic limb would be the induction of a collateral vascular network with angiogenic drugs treatment.Saxagliptin is a dipeptidyl peptidase-4 (DPP4) inhibitor that causes reductions in postprandial plasma glucose and might enhance eNO function. Some papers said the production of stromal derived factor-1 (SDF-1) was enhanced with DPP4 inhibition and increased production of endothelial progenitor cells in patients with type 2 diabetes mellitus. However, the role of DPP-4 inhibitors in angiogenesis is still unknown. Methods: Hind-limb ischemia modelEight-week-old male STZ-induced diabetic mice received vehicle (0.5% carboxymethyl cellulose), saxagliptin or glibenclamide administered by gavage every day. After 2 weeks of drugs treatment, unilateral hind limb ischemia was induced by excising the right femoral artery. Hind limb blood perfusion was measured with a Laser Doppler perfusion imager system before and after the surgery and then followed weekly. Capillary density in the ischemic limbAt 4 weeks after surgery, mice were sacrificed. Capillaries were identified by positive staining for CD31 and morphology. Capillary density was expressed as the number of capillaries per square millimeter. EPC mobilization in response to tissue ischemiaThe fluorescence-activated cell sorting Caliber flow cytometer was used to assess EPC mobilization. Circulating EPCs were considered to be from the mononuclear cell population and were gated with double positive for Sca-1 and Flk-1. Western blotting analysis of tissue superoxide and eNOs The gastrocnemius muscle tissue from nonischemic and ischemic hind limbs were isolated. Protein extracts underwent SDS-PAGE gel electrophoresis, were transferred to pure nitrocellulose membranes, and were probed with antibodies to eNOS, HO-1 and VEGF or α-tubulin. Results: Saxagliptin therapy promotes blood flow recovery in STZ-induced diabetic mice ( Figure 1). About the eNOs and HO-1 expression, It seems that saxaglitpin might enhances blood flow recovery after tissue ischemia in diabetic mice in an e-NOS-dependent and HO-1 dependent manner (Figure 2). Conclusions: The clinical implication of this study is to figure out the role of DPP-4 inhibitors in angiogenesis in diabetic microvascular complications . In the future, we may choose the optimal antidiabetic agents for the patients with microangiopathy of limbs.Introduction and Aims: Podocytes as part of the filtration barrier play an important role in the development of diabetic nephropathy (DN). Their numbers are significantly reduced in both type 1 and type 2 diabetic patients. Disturbances in insulin signaling accompanied by insulin resistance can lead to various intracellular events. Excessive flux through the hexosamine biosynthetic pathway and endoplasmic reticulum (ER) stress were related to insulin resistance. Autophagy serves as a protective response during ER stress through the degradation of unfolded proteins and damaged organelles. We hypothesized that glucosamine-induced insulin resistance in podocytes is associated with ER stress stimulation accompanied by induction of autophagy. Methods: Experiments were performed in primary rat podocytes cultured in the presence of 1-5 mM glucosamine (GlcN) for 3 and 5 days. Immunodetection methods were used to examine protein expression of ER chaperones (calnexin and Grp78) and LC3II protein, which is a commonly used marker of autophagy. Insulin-stimulated changes in glucose uptake were used to detect insulin resistance. To determine cell viability the colorimetric MTT metabolic activity assay was used. Results: The stimulating effect of insulin on glucose uptake in podocytes cultured for 5 days in the presence of 2 mM GlcN was abolished. The increase of calnexin and Grp78 proteins level was observed first after 3 days of incubation with 2 mM GlcN while after 5 days this effect was observed at 1 mM. Calnexin and Grp78 expression was further increased at 2 and 5 mM GlcN. Glucosamine treatment of podocytes for 5 days, activated autophagy, as indicated by increased LC3-II levels in the presence of 1, 2 and 5 mM GlcN. The same conditions reduced cell viability of podocytes. Conclusions: We found that impairment of insulin induction of glucose uptake into podocytes cultivated in the presence of glucosamine for long periods of time is associated with ER stress and autophagy induction.Introduction and Aims: Male gender predisposes to chronic kidney disease. We showed before increased circulating ACE2 and ACE in diabetic male mice. We studied gender differences and effect of diabetes and gonadectomy(Gdx) in circulating and renal ACE2 and ACE in STZ-induced mice. Methods: Study groups: Control and diabetic females (fCONT,fDB), males (mCONT,mDB) and Gdx males (mCONT+Gdx, mDB+Gdx). Gdx was performed 12 days after STZ injection. At the end of the study (19wks postSTZ) blood glucose(BG), body weight, kidney/body weight, blood pressure and urinary albumin excretion(UAE) were measured. Serum (s) ACE2 and ACE activity were determined by fluorometric assays. Kidney (k) ACE2 and ACE gene expression were quantified by RT-PCR and referred to GAPDH. Results: BG was significantly higher in diabetic males as compared to females during the first 12 weeks of follow-up. Gdx significantly reduced BG as compared to non-Gdx mice through the study. Serum ACE2 and ACE in fDB and mDB were increased as compared to controls (Table). Male control mice showed significantly higher sACE2 and sACE than females. Gdx resulted in significant reduction of sACE2(in both mCONT and mDB) and sACE(only in mDB). At kidney level, diabetes increased kACE2 but decreased kACE gene expression in both male and female mice. In addition, kACE was augmented in both control and diabetic male mice as compared to females. This increase was reduced by Gdx in control males. We found significant correlation between final BG and ACE2 and ACE in serum (r=0.6/r=0.5,p <0.01). Conclusions: Increased BG in diabetic mice was accompanied by higher circulating ACE2 and ACE activity in all diabetic groups. In addition, increased kidney ACE2 and decreased kidney ACE gene expression were observed in diabetic males and females. Gdx diminished BG, circulating ACE2 and circulating and renal ACE. These results suggest that, ACE2 and ACE alterations observed in diabetic male mice at serum and kidney level may be ascribed to a dysregulation of male sex hormones in type 1 diabetes.Introduction and Aims: Diabetes and hypertension independently contribute to kidney damage but often coexist, increasing the risk of diabetic nephropathy, a prime cause of end-stage renal disease. Increased production of reactive oxygen species (ROS) and overstimulated renin-angiotensin system (RAS) play an important role in the establishment and progression of kidney injury that is associated with both pathologies, though the mechanisms are not fully characterized. The renal medulla is very relevant for the control of blood pressure and a major contributor for renal dysfunction. We aimed to investigate the renal production of angiotensinogen (AGT), usually associated with increased angiotensin II concentration in the kidney, and ROS in diabetic normotensive and hypertensive rats. Methods: Diabetes was induced with streptozotocin (65 mg/kg, i.p.) in twelve week-old male Wistar and spontaneously hypertensive rats (SHR) rats; controls received vehicle. Three weeks later, intra-arterial systolic blood pressure (SBP), plasma and urinary AGT, renal oxidative stress and metabolic status were evaluated. Statistical interactions between groups of Wistar and SHR animals were evaluated by two way ANOVA followed by Tukey’s test for multiple comparisons. Values are means ± SEM. p < 0.05 was considered statistically significant. Pearson’s single regression analysis was used to estimate correlation between parametric data. Results: SBP was higher in SHR-controls than in Wistar-controls (200±6 vs 127±3 mmHg, respectively) and decreased in SHR-diabetics but not in Wistar-diabetics (143±8 and 122±6 mmHg, respectively). Diabetes increased urinary AGT excretion by more than 7-fold in both strains but decreased plasma AGT concentration by 40% in Wistar and 60% in SHR animals. Renal medullary H2O2 production was increased in diabetics (0.32 ± 0.04 vs 1.11 ± 0.10 nmol/mg protein for Wistar and 0.4 ± 0.05 vs 0.9 ± 0.14 nmol/mg protein for SHR) and was positively correlated with hyperglycemia (Wistar: r=0.7166, p<0.05; SHR: r=0.5763, p<0.05) and urinary AGT excretion (Wistar: r=0.8334, p<0.05; SHR: r=0.5620, p<0.05). Plasma AGT concentration was inversely correlated with urinary AGT excretion (Wistar: r=-0.8619, p<0.05; SHR: r=-0.8078, p<0.001) and positively correlated with SBP (Wistar: r=0.7312, p<0.05; SHR: r=0.7943, p<0.05). Conclusions: Our results show that hyperglycemia is accompanied by increased renal medullary H2O2 production that is correlated with the increased renal AGT. Interestingly, renal AGT seems to influence systemic AGT regulation.Introduction and Aims: Dipeptidyl peptidase-IV (DPP-IV) inhibition is currently being regarded as a promising strategy for diabetes management. However, its direct anti-enzymatic effect on kidney and its consequences are largely unknown. Methods: Normal chow-fed control mice, vehicle-treated high fat diet-induced obese mice, and DPP-IV inhibitor (LC15-0444)-treated high fat diet-induced obese mice were assessed for changes in various metabolic parameters, DPP-IV activity in target organs including kidney, and the parameters for renal damages. Results: After 3 months of treatment, the high fat-induced obese mice administered LC15-0444 at the dose of 3 mg/kg showed no differences in the levels of blood glucose, blood pressure, insulin resistance, and oxidative stress parameters both in kidney and adipose tissue compared with control obese mice. However, treatment with LC15-0444 significantly decreased levels of DPP-IV activity in serum, kidney, fat, and liver of the experimental mice. Furthermore, the mice treated with LC15-0444 showed significantly reduced albuminuria and the tendency to preserve creatinine clearance. In addition, renal histologic examinations showed that the mice treated with LC15-0444 also showed a significantly decrease in glomerulosclerosis and expression of pro-fibrotic markers. In support for these results, a subsequent in vitro study showed that LC15-0444 ameliorated obesity-induced increases in protein expressions of HMGB1, NF-κB, and P-ERK1/2 in renal cortical tissues. Conclusions: In conclusion, Administration of LC15-0444 reduced obesity-induced Increases in enzymatic activity of DPP-IV in the kidney as well as in plasma, and ameliorated renal damages independently of glucose-lowering effect. This renoprotection might be at least partly mediated by anti-inflammatory and anti-proliferative properties.Introduction and Aims: CD39 and CD73 are an ecto-enzyme that degrades extracellular nucleotides, such as ATP. Recent evidences indicate that CD39 and CD73 expression by regulatory T (Treg) cells contributes to the protective role in renal injury in an animal model of chronic renal injury. In addition, recent studies demonstrate the protective anti-inflammatory role of CD39/CD73 molecules pathway during renal hypoxia. However the role of CD39 and CD73 has yet not to be clarified in diabetic ESRD patients. This study was designed to evaluate the balance and role of CD39/CD73 of Treg cells in diabetic ESRD patients. Methods: Fifteen diabetic ESRD patients receiving hemodialysis and 15 healthy controls were recruited. Peripheral blood mononuclear cells were collected and stained with antibodies to CD4, CD25, FOXP3, CD39 and CD73. The balance of CD39/CD73 Treg cells were measured by using flow cytometry. Intracellular reactive oxygen species (ROS) generation was measured by flow cytometry using fluorescent dye, H2DCF-DA. The frequency of apoptotic cells was calculated by scoring annexin V-binding cells after back-gating of CD16/CD56 cells. Results: Diabetic ESRD patients had an increased frequency of Treg (HC vs ESRD, 2.57±0.3 vs. 10.04±3.1, p=0.0001) and CD39 cells (HC vs. ESRD, 11.66±2.5 vs. 33.72±6.9, p=0.01) and a decreased frequency of CD73 cells (HC vs. ESRD, 6.56±2.9 vs. 2.23±0.2, p=0.02). Intracellular ROS generation was significantly increased in diabetic ESRD patients (HC vs. ESRD, 8.84±2.6 vs. 58.25±11.0, p=0.01). In addition, ESRD patients showed a positive correlation between the frequency of ROS and both apoptosis and CD39 cells (apoptosis, r=0.696, p=0.04; CD39, r=0.458, p=0.04). Conclusions: Tregs isolated from diabetic ESRD patients lacked protective anti-inflammatory activity, suggesting that diabetic ESRD interferes with Treg functionality. The dysregulated balance of CD39/CD73 cells in diabetic ESRD patients provides new insights into an alteration of immunity and the pathogenesis of renal tissue injury in diabetic ESRD patients.Introduction and Aims: Diabetes mellitus causes damage in both heart and kidneys, leading to the cardiorenal syndrome that is associated with increased morbid-mortality. Additionally, it causes mitochondrial dysfunction; while function is responsible for several metabolism parameters. Although exercise training is a well described method to prevent DM damages, it is not clear how exercise training influences on mitofusin 2; that has been characterized as an important protein on mitochondrial metabolism. Also, exercise training is an interesting strategy for reversing the effects of mitochondrial dysfunction that might be through the regulation of mitochondrial protein transcription and biogenesis. Objective: To verify whether endurance exercise training could improve renal and cardiac parameters and increase mitofusin2 expression in cardiac myocytes in experimental streptozotocin (STZ) diabetes. Methods: Male Wistar rats were divided into 4 groups: control (C=8), control trained (CT=8), diabetic (D=8) and diabetic trained (DT=8). Diabetes (glycemia >250mg/dL) was induced by STZ (50mg/kg, i.v.). Trained groups were submitted to an exercise training protocol on a treadmill (8 wk). SBP, HR were analyzed using a data acquisition system (PowerLab). Renal parameters were evaluated using proteinuria (codas) and microalbuminuria (Elisa) measurements. The tissue weight/tibia length ratio was used to determinate the cardiac and kidney hypertrophic index. The expression of mitofusin 2 (mtf2), a mitochondrial transmembrane protein, was evaluated by Western blotting. Results: Exercise training improved renal parameters in the DT group showing a decreased urinary volume (DS: 153; DT: 106; CS: 10 and CT: 13 ml), proteinuria (DS: 46 ; DT: 29; CS: 18; CT: 7 mg/24h) and the microalbuminuria (DS: 8.7; DT: 4.7; CS: 1.5; CT: 2.3 mg/24h). The SBP was similar between groups (CS: 124, CT: 128; DS: 122; DT: 130 mmHg), however, the HR decreased in the DS (323bpm), while the DT (342 bpm) was increased é estatístico? Não é ao contrario?. Diabetes resulted in a reduction in the cardiac mass (23% decrease in the heart weight-to-tibia length ratio), on the other hand, the exercise training prevented this loss (DT=13 %) when compared to the control animals. Surprisingly, DM induced an increased in the kidney mass (DS: 28 %) and the exercise training did not influence on it (DT: 28%) when compared to control animals. The expression of mft2 was reduced in DS group and exercise training normalized this expression. Conclusions: Exercise training suppressed the progression of diabetic nephropathy and cardiac dysfunction; those improvements could be a result of the increased expression of mtf2, an important protein to the mitochondrial biogenesis pathway. These findings show that the exercise training is an important approach to avoid, both molecular and functional complications caused by diabetes.Introduction and Aims: Oxidative stress and inflammation are known to play a central role in the development of diabetic nephropathy. Febuxostat (Fx) is a novel nonpurine xanthine oxidase (XO)-specific inhibitor for treating hyperuricemia. The reduction in oxidative stress by administering the XO inhibitor has been shown to slow the progression of renal dysfunction. In addition, the renoprotective effects of an XO inhibitor have been suggested as uric acid lowering effect and anti-inflammatory effect. In this study, we investigated whether Fx could attenuate diabetic kidney injury and impart renoprotective effects, including anti-oxidative stress and anti-inflammatory mechanisms. Methods: Male Sprague-Dawley rats were divided into three groups: normal, vehicle-treated diabetes (DM), and febuxostat-treated diabetes (DM+Fx). We administered 5mg/kg of Fx to experimental rats for 7 weeks. Results: Urinary albuminuria was significantly reduced in Fx-treated diabetic rats. Quantitative analysis showed that hepatic XO and XDH activity was increased in the DM group (DM and DM+Fx groups) but reduced after treatment with Fx. Urine 8-OHdG concentrations and renal cortical nitrothyrosine also indicated reduced oxidative stress in the DM+Fx group relative to the DM group. We also observed a greater number of ED-1 stained cells in the glomerulus and tubule of diabetic renal tissue compared to normal; after administration of Fx, ED-1 stained cell count decreased. Finally, diabetic rats showed increased mRNA expression of inflammatory genes(E-selectin and VCAM-1), inflammation inducible enzymes(COX-2), and inflammatory mediators(ED-1 and NF-kB): after administration of Fx, these showed decreased significantly. Conclusions: Febuxostat ameliorates the diabetic renal injury such as albuminuria. Renoprotective effects of Fx may attenuate the inflammatory and oxidative stress mechanisms of renal damage in diabetes by inhibiting XO and XDH activity.Introduction and Aims: Mycophenolate mofetil (MMF) is a commonly used anti-lymphocyte drug with immunosuppressive/anti-inflammatory properties and has been used in recent years to prevent glomerular injury. It is a reversible inhibitor of inosine monophosphate dehydrogenase in purine biosynthesis which is necessary for the growth of T cells proliferation. Proinflammatory T helper 1 (Th1) and T helper 17 (Th17) cell subsets have been associated with the pathogenesis of multiple autoimmune diseases. We already reported that CD4 T cell is increased in diabetic kidney. However, the role of Th1 and Th17 cells in the development and progression of diabetic nephroapathy remains unknown state. In this study, we examined the hypothesis that MMF attenuates diabetic kidney injury by depression of renal T-cell proliferation and related cytokine. Methods: Streptozotocin (STZ)-induced diabetic mice were treated with 30mg/kg daily MMF during 3 to 20 weeks of diet. Body weight, kidney weight, fasting blood glucose, and glycosylated hemoglobin (HbA1c) were measured at the time of sacrifice. Twelve-hour urinary albumin-creatinine ratio and HbA1c were measured by immunoassay. To assess renal tissue damage, PAS-stained kidney sections Kidney sections were stained with PAS and evaluated for the presence of mesangial matrix expansion. IFN- IL-17 production of kidney infiltration CD4 T cells was investigated in kidney mononuclear cell by flow cytometry. Results: The HbA1c level were equally elevated with or without MMF in STZ-induced mice. Twelve-hour urinary albumin excretion increased markedly in diabetic mice, but decreased urinary albumin excretion in MMF-treated diabetic mice. Blood neutrophil and WBC counts showed mild reduction by MMF-treatment. In flow cytometry of kidney mononuclear cell, diabetic mice showed increase of IFN-γ for Th1 cells and IL-17 for Th17 cells from 8 weeks. MMF reduced the production of a number of T-cell cytokines as IFN-γ for Th1 cells and IL-17 for Th17 cells at 8 weeks. However, IFN-γ production was not inhibited by MMF, whereas IL-17 production was inhibited by MMF until 20 weeks. Conclusions: Our study results indicate MMF attenuates renal inflammation and glomerular injury by depression of renal IL-17 production in diabetic mice.Introduction and Aims: Diabetic Nephropathy (DN), a progressive renal disease, affects approximately 20-40% of patients with type 2 diabetes mellitus, being the most common cause of end-stage renal disease. This pathology is characterized by excessive accumulation of extracellular matrix, thickening of glomerular basement membrane and cell hypertrophy, which ultimately progress to glomerulosclerosis and tubulointerstitial fibrosis. Increasing evidences suggest that hyperglycemia-induced inflammatory processes and apoptotic cell death play an important role in the development and progression of DN. Previous studies from our group have shown that sitagliptin, an oral antidiabetic agent, corrects the glycemic dysmetabolism, and inflammation, and exerted beneficial effects on the blood-retinal barrier integrity in Zucker Diabetic Fatty (ZDF) rats, a model of type 2 diabetes. Sitagliptin is a dipeptidyl peptidase IV (DPP-IV) inhibitor that has shown to improve glycemic control by stabilizing the active incretin hormones and therefore increasing insulin secretion in T2DM patients. In this context, the aim of this study was to evaluate whether sitagliptin can exert beneficial and protective effects on the kidney in ZDF (fa/fa) rat, a tissue that is commonly affected by diabetic microvascular complications. Methods: Obese diabetic ZDF (fa/fa) at 20 weeks of age were treated with sitagliptin (10 mg/kg bw/day) during 6 wks. Insulin, glucose and HbA1c were evaluated in plasma, serum and total blood, respectively. The mRNA and/or protein content of CD26, GLP-1, TNF, IL-1β, BAX, Bcl-2 and Bid was evaluated in the kidney by RT-PCR and/or western blotting. The protein distribution was evaluated by immunohistochemistry. Results: Sitagliptin therapy was able to prevent the decrease in GLP-1 levels in the diabetic kidney. Furthermore, treatment with sitagliptin induced a significant decrease in the levels of the proinflammatory cytokines IL-1β and TNF, accompanied by a decrease in BAX/Bcl-2 ratio and in pro-apoptotic protein Bid, in the kidneys of diabetic animals. Conclusions: Overall, we can conclude that sitagliptin treatment has beneficial effects in the inflammatory and apoptotic state on the diabetic kidney in this animal model, which could play a key role in the prevention of DN evolution. This work was supported by the Foundation for Science and Technology, Portugal (PEst-C/SAU/UI3282/2011, PEst-C/SAU/UI3282/2013 and COMPETE).Introduction and Aims: Epithelial to mesenchymal transition (EMT) is process by which renal epithelial cells undergo a phenotypic conversion that give rise to the matrix producing myofibroblast in response to high glucose level. Several lines evidence suggests that high glucose not only activate ROS but also release Ang II in mesangial cells linked to EMT. Betanin is a natural pigment isolated from fruits of Opuntia elatior Mill (Cactaceae) with antioxidant properties used as a food colorant.The study aims to perform in-silico and invitro evaluation of betanin as an ACE inhibitor and antoxidant. Similarly, to determine whether betanin could attenuate high glucose induced EMT in the NRK-52E cell line by inhibiting ACE and oxidative stress. Methods: The in-silico docking study was performed by using target enzyme ACE (PDB: 1086) for ACE inhibitor activity. The in-vitro ACE inhibitor activity was performed by estimating the conversion of hippuric acid from HHL using RP-HPLC. Assessment of cell viability, Intracellular ROS and SOD and Cell morphology and wound healing assays were performed on NRK 52E cell line. Results: Docking study showed that Betanin have high binding energy of special precision −6.89 kcal/mol and extra precision -11.02. Betanin dose dependently inhibited the ACE activity compared with lisinopril. High glucose induces ROS in NRK 52E Cells which was inhibited by betanin. NRK-52E cells displayed typical cobblestone morphology of epithelial cells when the cells were grown in culture medium. Exposure to high glucose for 24 h resulted in a phenotypic conversion from epithelial cells into spindle shaped fibroblast like cells. Pretreatment of betanin significantly lessened the high glucose-induced phenotypic conversion in NRK 52E. Conclusions: Experimental and clinical studies have suggested that angiotensin II and oxidative stress plays a critical role in the pathogenesis and progression of diabetic complications. Accumulating evidence indicated that high glucose level contribute to the development of renal fibrosis in diabetes.Our data indicate that high glucose activates angiotensin II and increases oxidative stress that leads to EMT in NRK-52E cells. betanin exhibited protective action against high glucose-induced EMT of NRK-52E cells by inhibiting ACE and scavenging ROS, which might be beneficial in the prevention of renal fibrosis in diabetic nephropathy. Molecular Modeling Study of betanin on ACE (PDB : 1086) Sr. No. Compound SP score XP score 1 Lisinopril -6.7690 -6.4954 2 Betanin -6.892 -11.021Introduction and Aims: Autophagy is a rapidly advancing field of research in diabetic kidney disease. It attenuates glomerular damage through protection of glucose-induced podocyte injury. However its role in diabetic renal tubules remains unclear. This study aimed to examine the roles of autophagy in diabetic tubulopathy. Methods: Diabetic mice were induced by high fat high sucrose (HFHS) diet. H&E stain was employed to delineate the pathology in diabetic mice kidneys. Renal tubular cell (HK2) culture models were used to investigate the regulating mechanisms of autophagy. Transmission electron microscope (TEM) was used to demonstrate the formation of autophagosome. Western blot and flow cytometry were used to assay the LC3-II expressions. Pharmacology inhibition and RNAi were used to investigate the role of autophagy in HK2 cells cultured in the presence or absence of glucose. Mitochondrial morphology were stained by mitotracker and analyzed by confocal microscopy. TUNEL assay was used to examine the cellular apoptosis in glucose-treated HK2 cells Results: HFHS diet led to vacuolization and thyroidisation of the renal tubules in mice. Glucose caused mitochondrial fragmentation and cellular apoptosis in HK2 cells. Autophagy was activated, as indicated by the enhanced LC3-II expression, in glucose-treated cells. TEM demonstrated the formation of autophagosome in glucose-treated HK2 cells. When autophagy was inhibited, either by the inhibitor, 3-MA, or by siATG5, tubular cells were more susceptible to glucose-induced mitochondrial fragmentation and cellular apoptosis. Conclusions: Our results characterized the pathology of diabetic tubulopathy and suggested that glucose leads to mitochondrial fragmentation and cellular apoptosis in renal tubules. We also offered evidence that autophagy acts as a protective mechanism in diabetic tubulopathy.Introduction and Aims: Diabetic mice, with different Haptoglobin (Hp) genotype (1-1, 2-2) have a different susceptibility to developed Diabetic Nephropathy (DN). Hp 2-2 diabetic mice have impaired hemoglobin clearance and increased iron release in kidney proximal tubules (PCT); compared with Hp 1-1 diabetic mice. Patients and mice with chronic kidney disease (CKD) have decrease expression of the antioxidant protein ,renal klotho, that accompanied with active 1,25-dihydroxy vitamin D deficiency. Our research proposed a molecular mechanism explaining the influence of Hp genotype and klotho expression on active vitamin D deficiency in Diabetic Nephropathy patients. specific aims:1. To explore if the increased iron depositions in the renal PCT of Hp2-2 genotype as well as the decrease in renal klotho, generate a pro-oxidative environment, leading to high level of renal PCT apoptosis. 2. To explore if the elevated renal apoptosis interferes with vitamin D activation by 1-alpha hydroxylase in the renal PCT. 3. To investigate the role of active vitamin D supplementation in preventing the initiation and progression of kidney damage in Hp 2-2 DN mice. Methods: We subjected slides from kidney biopsies of DN patients and mice with different Hp genotype (1-1, 2-2) to Immunohistochemistry staining of iron,klotho, cleaved caspase 3, VDR and 1-alpha hydroxylase by using specific antibodies. Blood samples subjected to laboratory evaluation and different ELISA assays. Results: There was increased iron depositions in the renal PCT of Hp2-2 DM patients and mice compared with Hp1-1 DM genotype that corellateswith decrease levels of secreted klotho.Furthermore, Hp 2-2 genotype correlates with increased expression of active caspase-3 and decrease levels vitamin D receptor (VDR). Conclusions: These results provide insights into genetic predisposition to oxidative stress and the relationship between Renal PCT and macrophage apoptosis that may explain advanced sclerosis and active vitamin D deficiency in human Hp2-2 kidney biopsies. We hope that our observations will be translated to the clinical practice and will lead to developing a pharmacogenomics clinical approach to deal with DN complications by selective administration of vitamin D.Introduction and Aims: Diabetic Nephropathy (DN) is a clinical complication of Diabetes. Up to date there is still no cure for DN and this condition is today the major cause of End Stage Renal Disease (ESRD). As widely acknowledged, miRNA expression is strictly connected to protein synthesis as each miRNA modulates the expression of hundreds of targets. Aim of our study was to identify a pattern of differentially expressed miRNAs in kidneys of patients with DN. Methods: Using a microarray-based approach, we measured miRNA expression in FFPE kidney tissue coming from 8 patients with biopsy-proven DN, increased proteinuria and decreased glomerular filtration rate (GFR), and 4 control subjects with no signs of nephropathy. Results: 73 miRNAs were differentially expressed in DN; 18 were confirmed as enriched using qPCR. The table below shows a list of all the known mRNA targets for our qPCR validated DN-specific miRNAs. Known Targets hsa-miR-101 EZH2, PTGS2, STC1 hsa-miR-106b BCL2, BCL2L11, CCND1, CDKN1A, ESR1, IL8, PKD2, PPARG, PTEN, RB1, STAT3, TP63, TUSC2, VEGFA hsa-miR-126 VCAM1, VEGFA hsa-miR-140-5p SMAD3, VEGFA hsa-miR-141 CLOCK, CTNNB1, TGFB2, ZEB1 hsa-miR-146a BRCA1, CFH, CHUK, CXCR4, IFNA1/IFNA13, IL10, IL12RB2, IL1R1, IL1RAP, IL8, IRF5, NOS2, STAT1, TLR1, TLR10, TLR4, TLR9 hsa-miR-148a NR1I2 hsa-miR-15a ARHGDIA, BCL2, CA12, CCND1, CCND3, CHORDC1, EGFR, F2, FGFR1, HMOX1, HSP90B1, HSPA1A/HSPA1B, JUN, LAMC1, MAP2K1, NFIA, NPR3, PTGS2, VEGFA, WT1, ZYX hsa-miR-185 AKT1 hsa-miR-19a BCL2L11, CCND1, CTGF, ESR1, PTEN, THBS1 hsa-miR-214 PTEN hsa-miR-29a CD276, HMGN3, LAMC1, PIK3R1, SPARC hsa-miR-429 PPM1F, ZEB1The joint miRNA/mRNA target analysis also revealed three proteins whose reduction could be crucial in the context of glomerular filtration: vascular endothelial growth factor (VEGFA), essential for podocytes survival and preservation of the glomerular microvasculature; membrane-associated guanylate kinase inverted 2 (MAGI2), a newly discovered component of the slit diaphragm; and sulfatase 1 (SULF1), an enzyme that modulates the interaction of heparin sulfate glycosaminoglycans with growth factors, cytokines, chemokines and adhesion molecules. Conclusions: The further validation of these miRNAs and their targets respectively in biofluids and kidney tissue could bring to the identification of novel disease biomarkers and mechanisms involved in DN pathogenesis and progression.Introduction and Aims: The use of high-fructose corn syrup (HFCS, blend of 50-55% fructose (F) and 42-45% glucose (G)) as a sweetener in the U.S. coincided with increase of the prevalence of obesity and chronic kidney disease. A higher F content in HFCS than in sucrose was suspected as a potential explanation, given that diets excessively rich in F were found to cause metabolic syndrome, as well as glomerular hypertension, tubulointerstitial injury and decreased renal blood flow in animals. We aimed to compare long-term effects of sucrose vs. F and G in proportions corresponding to HFCS on body weight, visceral fat mass, tissue triglyceride (TG) content, and basic kidney function parameters in Sprague-Dawley (S-D) rats, with daily F intake close to that in general population. Methods: 24 6-week old male S-D rats were assigned to 34 weeks of unlimited access to 5% solutions of sucrose (8 rats, S group), 55%F and 45%G (8 rats, 55F/45G) or tap water (8 rats, W) and standard chow containing ~60% carbohydrates and no F. Fluid, chow, F, and total energy intakes, as well as weight changes, retroperitoneal fat mass, serum and liver TG levels, creatinine clearance and urine protein/creatinine ratio were compared between the groups. Results: Weight gain, total energy intake, and TG levels, as well as glomerular filtration and proteinuria did not differ between study groups. On the other hand, retroperitoneal fat mass was ~40% greater in rats consuming either of the sugar solutions than in W group (table). Conclusions: Unrestricted intakes of solutions of sucrose or HFCS-like F&G blend for 34 weeks do not significantly increase body weight or serum and liver TG content, nor do they affect kidney function in S-D rats. However, their consumption leads alike to increased adipose accumulation in the retroperitoneal depot, indicative of an increase of visceral adiposity with F consumption from either of the most common dietary sources. S(n=8) 55F/45G(n=8) W(n=8) Final body weight and body weight gain/day [g] 569.2±37.71.58±0.17 599.6±44.31.69±0.15 566.1±49.81.54±0.20 Fluid intake/day [ml] and chow intake/day [g] 74.9±19.0*19.3±2.8* 75.9±16.4*17.1±2.4* 46.0±10.323.0±3.3 Total energy intake/day and energy from fructose/day [kJ] 292.3±34.128.9±7.3* 263.7±25.732.1±6.9* 276.2±39.60 Fasting serum TG [mmol/L] and liver TG content [mg/g] 1.00±0.53.66±1.38 0.81±0.463.31±0.49 0.66±0.143.17±1.05 Retroperitoneal fat mass [g] and retroperitoneal fat mass/body weight [%] 11.3±3.5**2.0±0.5* 11.9±3.7*1.9±0.3* 8.1±2.81.4±0.4 Creatinine clearance [ml/min/kg bw] 4.39±1.47 3.51±1.07 4.63±1.93 Urine protein/creatinine ratio [g/g] 1.74±1.82 1.25±0.93 0.92±0.34 Mann-Whitney U-test; *P<0.05 vs. W; **P=0.05 vs. WIntroduction and Aims: Whereas ACE2 deletion worsens kidney injury, its amplification ameliorates diabetic nephropathy. The effect of gonadectomy(Gdx) in diabetic ACE2 knockout (ACE2KO) male mice has not been previously studied. Methods: We studied the effect of ACE2 deletion on systolic blood pressure (SBP), glomerular filtration rate(GFR), glomerular tuft area(GTA), mesangial index(MI), podocyte number (%POD) by Wilms Tumor staining, glomerular cellularity, and serum(s) and kidney(k) ACE expression in c57bl/6 streptozotocin(STZ)-induced male mice and their respective controls. We also evaluated the effect of gonadectomy in control(CONT) and diabetic(DB) ACE2KO mice. Mice were followed-up for 19 weeks after induction of diabetes with STZ injection. Citrate was administered as a vehicle. Study groups: Wild-type(WT)-CONT, ACE2KO-CONT, ACE2KO-CONT + Gdx, WT-DB, ACE2KO-DB, ACE2KO-DB + Gdx. Results: ACE2KO diabetic mice had increased SBP, GFR, and MI as well as decreased %POD as compared to diabetic WT. ACE2KO-DB + Gdx group showed lower values of these measured parameters excepted for augmented %POD. sACE was increased in WT and ACE2KO DB mice. Gdx decreased sACE in both CONT and DB ACEKO mice. kACE was reduced by diabetes in terms of enzymatic activity, protein and mRNA. kACE was also decreased in ACE2KO-CONT mice as compared to WT-CONT. kACE activity positively correlated with protein (r=0.7; p<0.001) and gene expression (r=0.481, p=0.005). Conclusions: Diabetic ACE2KO showed increased blood pressure and accentuated glomerular injury as compared to diabetic WT. These alterations were prevented by gonadectomy. Diabetes and ace2 deletion supposed increased circulating ACE but decreased renal ACE expression. Thus, the alterations observed in this model of diabetes may be related to a modulation of ACE at serum and kidney level. This regulation may be mediated, at least in part, by AngII accumulation.Introduction and Aims: The aging process is a complex phenomenon that promotes deleterious changes in cells and tissues, being responsible for the increased morbidity and mortality of these patients. Oxidative stress may change under several conditions, such as variations in glucose levels; it is involved in the aging process, and may result in cellular damage, also contributing to insulin resistance (IR). Animals supplemented with fructose (F) have been used as a well-established experimental model of IR. The aim of this study was to verify the renal and hepatic oxidative stress of senescent rats, with insulin resistance induced by high fructose intake. Methods: We utilized female Wistar rats with 3 (young) or 22 (aged) months of age, allocated in 4 groups: YC (young control), SC (senescent control), YF (young fructose) and SF (senescent fructose); n=5 for each group. The animals received standard chow (Purina ) ad libitum. The groups SC and YC received water; SF and YF received fructose 10% in the drinking water, both ad libitum. After 12 weeks of supplementation with high fructose or vehicle (water), the animals were sacrificed by decapitation and blood, liver and kidneys were collected. In the blood we measured nitric oxide (NO, µmol/mg protein), serum oxalacetic transaminase (SGOT, U/mL), serum pyruvic transaminase (SGTP, U/mL) and thiobarbituric acid reactive substances (TBARS nmol/mg protein), an indirect indicator of lipoperoxidation. In the kidney and liver tissues, we measured NO (by chemiluminescence with NOA™, a gold standard method for NO) and TBARS (by colorimetric method). Results are expressed as mean±SEM, analyzed by one-way ANOVA with Dunns post-test or Student t-test when needed; significance for p<0.05. Results: The values of fasting plasma glucose (mg/dL) showed significant difference between SF and SC (110±6 vs. 97±6), characterizing a fructose-induced IR. Our data indicated no significant changes in the serum, kidney or liver NO, neither in SGOT, among the groups. SGPT in SF was increased when compared with YF (72±6.5 vs. 49±1.6). TBARS in the kidney increased by age and by F administration in young rats: SC versus YC (0.82±0.1 vs. 0.55±0.1) and YF versus YC (0.95±0.1 vs. 0.55±0.1). In the liver, aging or F also increased TBARS: SC versus YC and YF versus YC (6±0.3 vs. 5±0.2). F in senescent rats decreased TBARS in both tissues. Histological analysis of liver showed that only SF presented apoptosis and some steatosis. Conclusions: In summary, our preliminary data show the effects of fructose on insulin resistance and TBARS, in the kidney and liver of female rats. In aged compared to young animals, we observed increased levels of kidney TBARS, a known marker of oxidative stress. Fructose administration increased TBARS in young female rats, both in the kidney and liver. These results need further attention to target preventive measures against tissue damage caused by aging.Introduction and Aims: It has been previously demonstrated that renin-angiotensin system (RAS) blockade delays chronic kidney disease progression and, thus, RAS is known to play a key role in diabetic nephropathy. In this sense, we work with the non-obese diabetic (NOD) mice, a strain which spontaneously develops autoimmune diabetes, mimicking type 1 diabetes in human. RAS in NOD mice is not well described. The aim of this study is to characterize the enzymatic activities of angiotensin converting enzymes (ACE and ACE2) in this strain. Methods: ACE and ACE2 enzymatic activities were studied in serum, renal cortex, lung, heart, liver and pancreas at 40 days after diabetes onset in NOD and in their control NOR (non obese resistant) mice by using fluorometric enzymatic assays. Results: ACE enzymatic activity was increased in serum, lung and liver in NOD diabetic as compared to NOR non diabetic mice. There were no differences between studied groups in renal, cardiac and pancreatic tissues. ACE2 was significantly increased in serum, kidney and heart samples of NOD as compared to NOR mice. However, lung, liver and pancreas did not show differences among studied groups (Table1). Conclusions: Circulating ACE and ACE2 activities are increased in NOD diabetic mice. Whereas ACE2 is also increased in kidney and heart, ACE is increased in lung and liver tissue. These results showed that ACE and ACE2 are modulated in diabetic NOD mice. In addition, our results suggest that the source of circulating ACE and ACE2 might be from different tissues.
  • Identifier: System Number: LDLAlsidyva3346695
  • Publication Date: 2014-05-01
  • Physical Description: Electronic Resource

Searching Remote Databases, Please Wait