益气活血化湿方及其拆方对足细胞的保护作用

Protective Effect of Yiqi Huoxue Huashi Decoction and its Decomposed Formula on Podocytes

目的:本研究探讨益气活血化湿方及其拆方对嘌呤霉素氨基核苷(PAN)诱导的足细胞损伤的保护作用。方法:培养足细胞并鉴定,建立PAN诱导的足细胞损伤模型。实验分为对照组(Con)、模型组(Model)、环孢素组(CsA)、全方组(QF)、益气组(YQ)、活血组(HX)、化湿组(HS)。分别利用RT-PCR与Western Bloting分析各组细胞CD_(2)AP、α-actinin4 mRNA和蛋白表达水平。结果:(1)Synaptopodin阳性表达,确立稳定的细胞培养条件;(2)明确造模浓度及各组用药浓度;(3)与Con比较,Model组α-actinin4 mRNA与蛋白表达均升高(P<0.05),CD_(2)AP mRNA与蛋白表达均下调(P<0.05);与Model比较,各用药组均能下调α-actinin4 mRNA与蛋白表达(P<0.05),上调CD_(2)AP mRNA与蛋白表达;QF与HS均能下调α-actinin4 mRNA与蛋白表达(P<0.05),二者差异无统计学意义(P>0.05);QF与YQ均能上调CD_(2)AP mRNA与蛋白表达(P<0.05),下调α-actinin4蛋白表达(P<0.05),二者差异无统计学意义(P>0.05)。结论:益气活血化湿方及其拆方能够通过修复骨架蛋白CD_(2)AP和抑制α-actinin4表达从而发挥足细胞保护作用,其中益气方和化湿方分别在修复CD_(2)AP表达与抑制α-actinin4表达方面作用显著。

Objective:This study explored the protective effects of Yiqi Huoxue Huashi Recipe and its decomposed formulas on podocyte injury induced by puromycin aminonucleoside(PAN).Methods:Culture and identify podocytes,and establish PAN-induced podocyte damage model.The experiment was divided into Con,Model,CsA,QF,YQ,HX,and HS group.The expression levels of CD_(2)AP andα-actinin4 mRNA and protein in each group were analyzed by RT-PCR and Western Bloting.Results:(1)Synaptopodin was positively expressed,and stable cell culture conditions were established;(2)Clarify the concentration of the model and the concentration of each group;(3)Vs the Con,the expression ofα-actinin4 mRNA and protein in the Model was increased(P<0.05),and the expression of CD_(2)AP mRNA and protein was down-regulated(P<0.05);Vs the Model,each medication groupcan down-regulate the expression ofα-actinin4 mRNA and protein(P<0.05),and upregulate the expression of CD_(2)AP mRNA and protein;both QF and HS can down-regulate the expression ofα-actinin4 mRNA and protein(P<0.05),and there is no significant difference between the two(P>0.05);QF and YQ can up-regulate CD_(2)AP mRNA and protein expression(P<0.05),down-regulateα-actinin4 protein expression(P<0.05),no significant difference(P>0.05).Conclusion:Yiqi Huoxue Huashi Recipe and its disassembled prescriptions can play a protective role in podocytes by repairing the skeleton protein CD_(2)AP and inhibiting the expression ofα-actinin4.Among them,Yiqi and Huashi had significant effects on repairing CD_(2)AP expression and inhibitingα-actinin4 expression,respectively.