糖尿病大鼠肾组织中AKT的表达及意义.docx

糖尿病大鼠肾组织中AKT的表达及意义来源：创新医学网? 　　作者：石明隽,肖瑛,桂华珍,郭兵,张国忠　　作者单位：贵阳医学院病理生理学教研室，贵州贵阳 550004　　【摘要】目的观察Akt1和Akt2在糖尿病大鼠肾组织中的表达并探讨其是否参与糖尿病肾病(DN)发病过程。方法链脲佐菌素(STZ)诱发大鼠糖尿病(DM)，分为2、4、8、12、16 w和24 w组，每一时点均设相应的正常对照组。测定各组血糖、24 h尿蛋白、血肌酐(Scr)、肾脏指数。HE和PAS染色光镜观察肾脏病理改变。免疫组化方法检测肾皮质Akt1、Akt2、TGF-β1、α-SMA和FN的表达，Western印迹和RT-PCR法检测Akt1、Akt2蛋白及mRNA表达水平。结果 DM各组大鼠的血糖、24 h尿蛋白、血肌酐、肾脏指数均较正常对照组明显升高(P0.05,P0.01)，免疫组化显示DM各组TGF-β1、α-SMA和FN阳性染色较正常组明显增多(P0.01),以12 w增多最为明显，分别为19.67±2.73、9.50±3.45、22.17±2.79，免疫组化、Western印迹及RT-PCR显示DM各组Akt1及Akt2蛋白和mRNA表达较正常组明显增多(P0.01)。免疫组化显示Akt1与TGF-β1、FN蛋白表达水平呈显著正相关(r=0.694，r=0.532,P0.01)，Akt2与TGF-β1、FN蛋白表达水平呈显著正相关(r=0.789，r=0.743，P0.01)。结论 Akt1、Akt2蛋白在DM大鼠肾组织过度表达，提示PI3K/Akt信号通路可能参与了DN的发病机制。　　【关键词】 Akt1;Akt2;转化生长因子β1;α-平滑肌肌动蛋白;纤连蛋白;糖尿病肾病　　【Abstract】 Objective To observe the expression of Akt1 and Akt2 in renal tissues of diabetic rats and explore its effect in the development of diabetic nephrophy. Methods The diabetic rats induced by streptozotocin (STZ) were randomly divided into 2, 4, 8, 12, 16 and 24 w groups. Control group was installated at age-matched time points. Blood glucose, 24 h urine protein, serum creatinine (Scr) of rats were measured. The protein expression of Akt1, Akt2, TGF-β1, α-SMA and FN in renal cortex were detected by immunohistochemical staining. Western blotting and reversetranscriptase-PCR (RT-PCR) were employed to detect the protein and mRNA expression of Akt1 and Akt2. Results The levels of blood glucose, 24 h urine protein and Scr were increased remarkably in diabetic rats compared with those in control groups (P0.05, P0.01). The protein and mRNA of Akt1 and Akt2 were significantly up-regulated. The TGF-β1, FN and α-SMA protein were significant increased in DM rats as compared with those of the controls. Conclusions Akt1 and Akt2 gene and protein expression are up-regulated in the kidney of DM rats, and which may involve in the pathogenesis of diabetic nephropathy.　　【Key words】 Akt1; Akt2; TGF-β1; α-SMA; Fibronectin; Diabetic nephropathy　　PI3K/A