缬沙坦对小鼠足细胞GPSD核心蛋白在高糖环境下表达的影响

Effect of Valsartan on Expression of Mouse GPSD Core Proteins in High-glucose.

目的通过观察缬沙坦体外干预高糖环境培养的小鼠足细胞nephrin、podocin和CD2AP的表达,探讨RAS抑制剂治疗糖尿病肾病蛋白尿的机制。方法用不同浓度的缬沙坦干预高糖(25mmol/L的葡萄糖)培养24h后的小鼠足细胞,以RT-PCR方法检测干预前后足细胞nephrin、podocin和CD2AP mRNA的表达水平;以间接免疫荧光法检测2×10-5mol/L缬沙坦干预后nephrin、podocin蛋白的表达。结果①与对照组相比,高糖诱导的足细胞nephrin、podocin和CD2AP mRNA表达显著减弱(P<0.01);②缬沙坦能显著上调高糖诱导的足细胞nephrin、podocin和CD2AP mRNA的表达(P<0.05或P<0.01);③2×10-5mol/L缬沙坦能显著上调高糖对足细胞nephrin和podocin(P<0.05)蛋白表达的抑制。结论缬沙坦显著上调髙糖诱导的足细胞nephrin、podocin和CD2AP的表达,可能是其独立于降压作用以外的降低糖尿病肾病蛋白尿的机制之一。

Objective To explore the mechanism of angiotensin Ⅱ receptor 1 antagonist attenuating proteinuria of diabetic nephropathy by observing the effect of valsartan（an angiotensin Ⅱ receptor 1 antagonist） on expression of mouse GPSD core proteins（nephrin,podocin and CD2AP） in high-glucose.Methods Mouse podocyte were randomly cultured without and with glucose（25mmol/L） incubated for 24h.Podocyte exposured to high-glucose then were randomly incubated with different doses of valsartan（2×10-7,2×10-6 and 2×10-5 mol/L respectively）.The expression of nephrin,podocin and CD2AP mRNA was examined by RT-PCR.The expression of nephrin and podocin protein was detected by IIFT（indirect immunoflorescence technique）.Results ①Compared with the control group,the expression of nephrin,podocin and CD2AP mRNA cultured with 25mmol/L glucose significantly decreased（P〈0.05 or P〈0.01） after respectively incubated with 3 different doses of valsartan.②The expression of nephrin and podocin in high-glucose was significantly up-regulated by GPSD core proteins incubated with high dose valsartan（2×10-5mol/L）.Conclusion Expression of mouse podocyte nephrin,podocin and CD2AP inhibited in high glucose is significantly up-regulated by valsartan,which indicates that it may play an important role in attenuateing proteinuria of DN independent of its effects on blood pressure.