The Study Of The Renal Protective Effects Of Probucol On Experimental Diabetic Rats And Its Mechanism

The Study of the renal protective effects of probucol on experimental diabetic rats and its mechanismObjective:DN is one of the main reasons to cause end stage renal failure.Studies have proved that,the increasing generation of oxygen free radicals is important in generation and development of DN,which was related to the hyperglycemia in diabetic state.Probucol lowers both LDL and HDL cholesterol,it is also an antioxidants.This study is to investigate the changes ofα-SMA,CTGF,TGF-β,FN in STZ-induced diabetes mice;to investigate the renal protective effects of probucol on experiment diabetic rats and its mechanism.Methods:36 SD male rats were randomly devided into 2 groups:E(ordinary diet,10 rats);C(high-glucose-high-fat diet,26 rats).Datas were evaluated such as serum lipids,serum glucose,fasting serum insulin in time of 8 week;Then the diabetes model were established by injection of STZ at dose of 30 mg/kg in group C.23 rats become diabetic rats model whose random blood glucose was above 16.67mmol/l.Then all the rats were reassigned to 3 groups:E(ordinary diet,also that 10 rats);A(diabetic group) B(diabetes and probucol-treated group).After 8 weeks,24h urine was collected before the rats were sacrificed and 24h urinary microalbumin was examined.The left kidneys were weighted to calculate the kidney hypertrophy index.The blood samples were collected to determined blood glucose,serum lipids(TG,TC,HDL,LDL),Scr, BUN.The renal cortex were collected to measure activities of SOD,GSH-PX and concentrations of MDA.HE staining was used to detect the renal pathological changes by light microscope.TGF-β1,FN,α-SMA were detected with Immunohistochemistry analysis.The levels of CTGF,TGF-β1 andα-SMA mRNA were detected by reverse transcriptasepolymerase chain reaction(RT-PCR).In addition,the ultrastructure of glomerular podocytes was observed by transmission electronic microscopy.The dataes were analyzed by SPSS 13.0 system.Results:(1)After 8 weeks of high-glucose-high-fat diet,88.46%of group C models were successful.(2)Body weight:the weight of group C was significantly higher than E,HOMA-IR of C significantly lower than E,which were identified as characters of T2DM.(3)Blood lipid:The level of TG,TC,LDL-C of group A were significantly higher than that of group E(P＜0.05).The level of TC,LDL-C of group B were significantly lower than that of group A(P＜0.05).(4)24h UMA and KW/BW:The level of 24h UMA,KW/BW of group A was significantly higher than that of group E(P＜0.05);there was no significance between the level of KW/BW in group A and that in group B(P＞0.05).(5)SOD,GSH-PX,MDA:The activities of SOD,GSH-PX in group A was significantly lower than group E(P＜0.05);the level of MDA was significantly higher than E (P＜0.05).the activities of SOD,GSH-PX in group B was significantly higher than group A(P＜0.05);the level of MDA was significantly lower than A(P＜0.05).(6)Expression of CTGF,TGF-β1,α-SMA,FN in the kidney:Expression of TGF-β1,α-SMA,FN in group A and B was significantly higher than that of group E(P＜0.05); Expression of CTGF,TGF-β1 andα-SMAmRNA in group A and B was significantly higher than that of group E(P＜0.05).(7)Electron Microscopic Observation in the kidney:thickened basement membrane, mesangial proliferation and fusion of the podocytic processes were easily observed in group A;severity of the fusion of the podocytic processes in group B was between that of group E and group A.Conclusions:(1)We successfully prepared rat model of T2DM with high-glucose-high-fat diet plus small dose injection of STZ,which method is simple and has high success rate of model making.(2)In STZ-induced diabetes rats,we demonstrate that the decreased antioxidantion and increased oxidative stress in kidney may rise transformation and differentiation of Glomerular Epithelium Cells and interstitial cells,the expression of CTGF,TGF-β1,α-SMA,FN rise.Probucol may improve the activity of antioxidant enzyme,reduce the content of oxidant substance and the level of renal CTGF,TGF-β1,α-SMA,FN expression.Probucol may provide a renoprotective effect.(3)Many researches were limited to vitro and animal study model.There are few researches basing on patient and clinic.They call on researchers to explore furtherly.