他克莫司对2型糖尿病大鼠ANGPTL2在肾脏中表达的影响

Effect of Tacrolimus on ANGPTL2 in the Kidney of 2 Type Diabetes Rat Models

目的:通过观察他克莫司(tacrolimus,FK506)对2型糖尿病大鼠(T2DM)肾脏中血管生成素样蛋白2(ANGPTL2)表达的影响,探讨其对肾脏的保护机制。方法:50只SD雄性大鼠随机分为正常对照组(NC,10只)和实验组(40只),实验组予高糖高脂喂养联合小剂量链脲佐菌素(STZ 30 mg/kg),建立T2DM大鼠模型。成功模型(32只)再随机分为两组:糖尿病对照组(DM,16只)及他克莫司组(DT,16只)。他克莫司干预8周后,测体重、血压、肾重、血糖、血肌酐(Scr)、尿素氮(BUN)、24 h尿蛋白(UAL)的排泄量;镜下观察肾组织病理改变、免疫组化染色技术、实时定量PCR及Western blot法检测肾组织ANGPTL2、Nephrin、Podocin mRNA及蛋白表达的变化。结果:DM组24 h UAL排泄量及足细胞损伤明显高于NC组,同时ANGPTL2 mRNA及蛋白表达量明显升高(P均<0.05),Nephrin及Podocin的mRNA及蛋白表达量明显降低(P<0.05),DT组较DM组24 h UAL排泄量降低,足细胞损伤改善,ANGPTL2 mRNA及蛋白表达量降低(P<0.01),Nephrin及Podocin的mRNA及蛋白表达量升高(P<0.01),且ANGPTL2 mRNA及蛋白表达量与尿白蛋白呈正相关,其与Nephrin及Podocin的mRNA及蛋白表达量呈负相关(P均<0.01)。结论:他克莫司可通过下调ANGPTL2在肾脏的表达,保护足细胞。

Objective： To observe the effect of TacroIimus on the expression of Angiopoietin - like protein 2 （ ANGPTL2 ） in the kidney of diabetic rats and discuss its mechanism. Methods ：50 SD male rats were divided into normal control group （ NC, n = 10） and experimental group （n = 40） randomly. The experimental group was fed with high sugar -fat diet and given a low dose streptozocin （STZ 30 mg/kg） established type 2 diabetic model rats, and then the successful induced models randomly divided into 2 groups： diabetes control group （DM） and tacrolimus group （DT）. Weight, blood, pressure, blood, glucose, serum creatinine （Scr） , blood iJrea nitrogen （BUN） 24 hour urinary albumin （UAL） and renal histomorphology were observed after Drug intervention for 8 weeks, the expression of ANGPTL2 ,Nephrin and Podocin in renal tissue were detected by immunohistochemisty, real - time PCR and Western blot. Results： UAL in DM group were obviously higher , compare to the NC group （ P 〈 0.05 ）, and the mRNA and pro- tein of ANGPTL2 distantly was upregulated compared with NC group （P 〈 0.05 ）, the mRNA and protein of Nephrin and Podocin were significantly decreased （P 〈 0.01 ） ; Meanwhile UAL was significantly lower （P 〈 0.01 ）. Compared with DM group, and the mRNA and protein of ANGPTL2 downregulated （P 〈 0.01 ）. Neverthless, the mRNA and protein of Nephrin and Podocin were significantly upregulated. Conclusion：These results indicate that Tacrolimus could protects the diabetic kidney through decreasing the expression of ANGPTL2.