黄芪菟箭合剂对高糖诱导的足细胞损伤及凋亡的影响

Effect of Huangqi Tujian decoction on injury and apoptosis of podocytes induced by high glucose

目的探讨黄芪菟箭合剂(HTD)对高糖诱导足细胞损伤及凋亡的影响。方法体外培养永生化的肾小球足细胞,待足细胞分化完全后,用不同浓度HTD(0.25,0.5,0.75,1,1.25,1.5 mg/ml)刺激足细胞24 h,用乳酸脱氢酶释放法(LDH法)判断细胞的损伤程度,进而筛选HTD的最佳浓度,并观察其对足细胞形态的影响。随机将分化的足细胞分为对照组、高糖组和黄芪菟箭合剂组(HTD组),通过Western blotting和免疫荧光分析检测足细胞标志蛋白Nephrin和ZO-1以及足细胞凋亡相关蛋白Caspase和NF-κB的表达水平。结果HTD的最佳给药浓度为0.75 mg/ml,其不明显改变足细胞形态。与对照组相比,高糖组标志蛋白Nephrin和ZO-1表达水平显著降低(P<0.05),凋亡相关蛋白Caspase和NF-κB的表达显著升高(P<0.05)。而与高糖组相比,HTD组Nephrin和ZO-1表达水平显著升高(P<0.05),而凋亡相关蛋白Caspase和NF-κB的表达显著下降(P<0.05)。结论在糖尿病肾病中,高糖可以诱导肾小球足细胞的损伤,与足细胞的凋亡有关,而HTD可干预高糖刺激所造成的足细胞损伤,并减少足细胞的凋亡。

Objective To investigate the effect of Huangqi Tujian decoction(HTD)on the injury and the apoptosis of podocytes induced by high glucose.Methods Immortalized glomerular podocytes were cultured in vitro.The differentiated podocytes were stimulated with different concentrations of HTD(0.25,0.5,0.75,1,1.25,1.5 mg/ml)for 24 h,and then the cell damage was assessed by the lactate dehydrogenase(LDH)assay to screen the optimal concentration of HTD,and the effect of HTD on podocyte morphology was observed.The differentiated podocytes were randomly divided into control group,high glucose group and HTD group.The expression le-vels of podocyte-related proteins Nephrin and ZO-1 and podocyte apoptosis-related proteins Caspase and NF-κB were detected by Western blotting and immunofluorescence analysis.Results The optimal concentration of HTD was 0.75 mg/ml,and the podocyte morphology did not change under this concentration.Compared with control group,the expression levels of Nephrin and ZO-1 proteins were significantly decreased in high glucose group(P<0.05),while the expression levels of apoptosis-related proteins Caspase and NF-κB were significantly increased(P<0.05).Compared with high glucose group,the expression levels of Nephrin and ZO-1 were significantly increased in HTD group(P<0.05),while the expression levels of apoptosis-related proteins Caspase and NF-κB were significantly decreased(P<0.05).Conclusion High glucose can induce the glomerular podocyte injury in diabetic kidney disease(DKD),which was related to the podocyte apoptosis.However,HTD can intervene podocyte injury incluced by high glucose and reduce the podocyte apoptosis.