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目的研究复方仙草颗粒对人肾足细胞的保护作用。方法在体外培养人肾足细胞,实验分为4组:阿霉素组、雷公藤甲素组、地塞米松组、复方仙草颗粒组。采用MTT法检测细胞增殖,荧光技术观察细胞骨架的形态变化,Realtime PCR法检测细胞内nephrin、CD2AP mRNA的表达,蛋白质印迹技术观察细胞内nephrin、CD2AP的蛋白表达水平,流式细胞仪观察细胞内ROS水平。结果与阿霉素组相比,复方仙草颗粒组人肾足细胞增殖速度明显增加,且具有一定的量效性和时效性;细胞骨架F-actin排列有序,荧光增强;细胞内ROS水平明显降低,足细胞相关蛋白nephrin和CD2AP表达明显提高。结论复方仙草颗粒通过抑制ROS水平,上调足细胞相关蛋白nephrin和CD2AP的表达,维护细胞骨架,从而对人肾足细胞起到保护作用。
Objective To study the protective effect of Fufang Xiancao Granules on human renal podocytes. Methods Human renal podocytes were cultured in vitro. The experiment was divided into 4 groups: doxorubicin group, triptolide group, dexamethasone group and compound jelly group. The cell proliferation was detected by MTT assay. The morphological changes of cytoskeleton were observed by fluorescence assay. The expression of nephrin and CD2AP mRNA was detected by Realtime PCR. The protein expressions of nephrin and CD2AP were observed by Western blotting. ROS level. Results Compared with doxorubicin group, the proliferation rate of human renal podocytes in Compound Xiancao Granules group was significantly increased, and the amount of F-actin was orderly arranged and the fluorescence was enhanced. The intracellular ROS level Significantly reduced, podocyte-related protein nephrin and CD2AP expression was significantly improved. Conclusion Compound Xiancao Granules can protect human renal podocytes by inhibiting the level of ROS, up-regulating the expressions of podocyte-related proteins nephrin and CD2AP, and maintaining the cytoskeleton.