左归降糖益肾方对2型糖尿病肾病小鼠足细胞nephrin与podocin表达的影响  

Effects of Zuogui Jiangtang Yishen Decoction on Expression of Nephrin and Podocin in Podocyte of MKR Mice with Diabetic Nephropathy

作　　者：印红爱[1] 吴勇军[1] 喻嵘[1] 成细华[1] 陈聪[1] 罗文娟[1] 何溪沁冰 程莉娟[1] 

机构地区：[1]湖南中医药大学,湖南长沙410208

出　　处：《中国中医药信息杂志》2014年第3期53-57,共5页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金(81273753;81102593);湖南省自然科学基金(12JJ2048);湖南省教育厅科研基金(11C0964);湖南省大学生研究性学习和创新性实验计划项目(2013-168);湖南省重点学科-中西医结合基础和中医内科学(2012年)

摘　　要：目的观察左归降糖益肾方对MKR转基因2型糖尿病肾病(DN)小鼠足细胞nephrin、podocin表达的影响,探讨其作用机制。方法 8周龄MKR小鼠40只随机分为4组,空白组(A组)、DN组(B组)、DN+中药组(C组)、DN+糖适平+贝那普利组(D组)。B、C、D组给予高脂饮食联合右侧肾切除造模。手术4周后,各给药组给予相应药物,4周后处死小鼠。电化学法观察小鼠空腹血糖,全自动生化仪检测血肌酐(SCr)、尿素氮(BUN),ELISA法测定尿微量白蛋白(UmAlb),RT-PCR法检测nephrin、podocin mRNA表达,Western Blotting法测定nephrin、podocin蛋白表达,电镜观测足细胞形态结构。结果与A组比较,B组小鼠空腹血糖、SCr、BUN及UmAlb均明显升高(P<0.01),肾小球足细胞nephrin、podocin mRNA及蛋白表达明显下调(P<0.01)。给药后,与B组比较,用药组小鼠血糖、SCr、BUN及UmAlb明显下降(P<0.01),肾小球足细胞nephrin、podocin mRNA及蛋白表达明显上调(P<0.01,P<0.05)。电镜结果显示,给药组小鼠肾小球足细胞较B组明显改善。结论左归降糖益肾方通过恢复2型糖尿病肾病小鼠足细胞nephrin和podocin的表达而保护肾脏。Objective To investigate the effects of Zuogui Jiangtang Yishen decoction （ZGJTYS） on the expression of nephrin and podocin in podocyte of MKR mice with diabetic nephropathy （DN）, and explore its mechanism. Methods Forty MKR mice （8 weeks old） were randomly divided into four groups as follows：negative control group （group A）, DN model group （group B）, ZGJTYS group （group C） and positive control group （group D, Gliquidone and benazepri）. All mice from group B, C and D were received high-fat diet feed and unilateral nephrectomy. Four weeks after operation, all mice were received drug intervention, and four weeks later, all mice were put to death. The levels of UmAlb were observed by ELISA, the serum BUN and Cr by biochemical, and the FBG by electrochemical detection method. The nephrin and podocin mRNA expression were measured by RT-PCR and the protein expression by western blotting. The morphological structure changes of the podocytes were observed by transmission electron microscopes. Results As compared with group A, FBG, BUN, SCr and urine UmAlb in the mice of group B increased significantly （P〈0.01）, the expression level of nephrin and podocin mRNA and protein were markedly decreased （P〈0.01）. After intervention of drugs, all biochemical indicators above in the mice of group C and D significantly decreased （P〈0.01）, the expression level of nephrin and podocin mRNA and protein were markedly increased （P〈0.01, P〈0.05）, compared with group B. The renal pathological lesions of group C and D were significantly improved compared with group B. Conclusion ZGJTYS decoction exerts reno-protective effect via reinstating nephrin and podocin expression to repair the damaged podocyte.

关 键 词：糖尿病肾病 左归降糖益肾方 足细胞 NEPHRIN PODOCIN MKR小鼠 

分 类 号：R285.5[医药卫生—中药学]