摘要
目的:探讨丹参酮ⅡA对嘌呤霉素诱导肾足细胞骨架蛋白F-actin、synaptopodin损伤的作用及其可能的作用机制。方法:条件永生性人肾足细胞株AB8/13随机分为对照(control)组、嘌呤霉素氨基核苷(puromycin aminonucleoside,PAN)组、不同剂量(10,20,40,80μmol)丹参酮ⅡA干预(tanshinoneⅡA,TⅡA)组。采用细胞免疫荧光法检测各组细胞骨架蛋白F-actin、synaptopodin在足细胞内的分布及形态;通过DAPI染色法观察各组细胞核的变化;免疫印迹法检测各组细胞p-Rac1/Cdc42蛋白的表达量。结果:(1)F-actin、synaptopodin细胞免疫荧光染色:较control组,PAN组F-actin、synaptopodin结构紊乱,胞质回缩,细胞边缘褶皱,凋亡小体增多。各浓度TⅡA组上述情况较PAN组显著改善。(2)p-Rac1/Cdc42蛋白表达量,与对照组相比,PAN组p-Rac1/Cdc42蛋白表达量显著上调。与PAN组相比,TⅡA组p-Rac1/Cdc42蛋白表达量显著下调,且呈浓度依赖方式。结论:TⅡA可改善PAN诱导的足细胞骨架蛋白F-actin、synaptopodin的重排及结构紊乱,对足细胞具有一定保护作用,其机制可能与下调Rac1/Cdc42磷酸化水平,减少足细胞丝状伪足、层状伪足形成,从而稳定PAN诱导的细胞骨架变构有关。
Objective： To observe the protective effect of Tanshinone ⅡA（ TⅡA） on puromycin aminonucleoside（ PAN）- induced podocyte injury,and to explore its potential mechanism. Methods： Conditionally immortalized human podocytes were treated with culture medium（ control）,PAN,and Tanshinone ⅡA（ 10,20,40,80 μmol） plus PAN. The distribution of F- actin and synaptopodin were studied by immunofluorescence,and the expression of p- Rac1 / Cdc42 was measured by Western Blot. Results：Immunofluorescence showed that PAN induced disruption of F- actin and synaptopodin,and TⅡA appeared to improve the disruption. Podocytes treated with PAN caused an increase in p- Rac1 / Cdc42 expression,while treated with TⅡA decreased the expression of p- Rac1 / Cdc42. Conclusion： These finding suggest that TⅡA may protect podocyte from actin cytoskeleton disruption or reorgnisation through suppressing the phosphorylation of Rac1 / Cdc42.
出处
《中国中西医结合肾病杂志》
2016年第3期221-223,I0005,I0006,共5页
Chinese Journal of Integrated Traditional and Western Nephrology
基金
国家自然科学基金青年科学基金资助项目(No.81202671)
