法舒地尔对血管紧张素II诱导的足细胞自噬的影响

肾脏病与透析肾移植杂志 ›› 2014, Vol. 23 ›› Issue (1): 42-47.

法舒地尔对血管紧张素II诱导的足细胞自噬的影响

出版日期:2014-02-28 发布日期:2014-02-25

Fasudil, a Rho kinase inhibitor, inhibits ANGII-induced autophagy in podocytes

Online:2014-02-28 Published:2014-02-25

摘要/Abstract

摘要：

[摘要] 目的：探讨法舒地尔对血管紧张素II（Ang II）诱导人足细胞（AB 8/13）自噬的影响。方法：体外培养人足细胞株，采用不同浓度的AngII(10-8～10-6mol/l)刺激人足细胞24h和固定浓度AngII (10-7 mol/l)刺激不同时间（0、6、12、24、36h），以及特异性自噬阻断剂3-MA(5mmol/l)和不同浓度（10-8～10-6mol/l）法舒地尔进行干预。免疫荧光法和Western印迹法检测自噬相关蛋白LC3B、Beclin-1的表达变化。结果：AngII可刺激足细胞LC3B、Beclin-1蛋白表达上调，且呈剂量、时间依赖性(p<0.05)，在浓度10-7mol/lAng II和时间24h时达高峰。3-MA能明显降低Ang II引起的LC3B蛋白表达(P<0.01)。单独的法舒地尔组对足细胞自噬没有影响；法舒地尔可明显下调AngII诱导的足细胞LC3B和Beclin-1蛋白表达(P<0.05)，10-7mol/l时下调最明显。结论：AngII可以诱导人足细胞自噬增强，3-MA和法舒地尔均可抑制此过程的发生，提示Rho激酶途径可能参与了AngII诱导的足细胞自噬过程。

关键词: 血管紧张素II, 自噬, Rho激酶, 足细胞, 3?甲基腺嘌呤

Abstract:

【Abstract】 Objective: To investigate the effect of fasudil (a specific Rho-kinase inhibitor) on angiotensin (Ang) II-induced autophagy in podocytes. Methodology: The conditionally immortalized human podocytes (AB8/13) were incubated with various concentrations of AngII (10-8 to 10-6 mol/l) for 24h or with 10-7 mol/lAng II for different times (0, 6, 12, 24 and 36 hours), together with or without autophagy inhibitor 3-methyladenine (3-MA) and various concentrations of fasudil (10-8 to 10-6 mol/l) in vitro. The protein expression of LC3B and Beclin-1 were detected by immunofluorescence and western blotting. Results: The proteins expression of LC3B and Beclin-1 were up-regulated significantly by Ang II in a dose- and time- dependent manner. The optimal concentration of Ang II was 10-7mol/l and the optimal time was at 24h in which AngII enhanced LC3B and Beclin-1 protein expression to the maximum . The stimulation of podocytes with Ang II increased LC3B and Beclin-1 expression that was prevented by 3-MA. The expression enhancement of LC3B and Beclin-1 by AngII were reduced significantly by fasudil in a dose- dependent manner, although fasudil alone does not induce autophagy. The optimal inhibitory concentration of fasudial was 10-7mol/l. Conclusion: The autophagy of podocytes can be induced by Ang II and inhibited by fasudil, which suggests that Rho-kinase pathway may mediate Ang II induced the autophagy of podocytes.

Key words: Angiotensin II, autophagy, Rho-kinase, podocytes, 3-methyladenine

高雪静，唐莎，尹仕伟，等. 法舒地尔对血管紧张素II诱导的足细胞自噬的影响[J]. 肾脏病与透析肾移植杂志, 2014, 23(1): 42-47.

GAO Xuejing, TANG Sha, YIN Shiwei,et al. Fasudil, a Rho kinase inhibitor, inhibits ANGII-induced autophagy in podocytes[J]. Chinese Journal of Nephrology, Dialysis & Transplantation, 2014, 23(1): 42-47.

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