脂联素球状结构域对高糖环境下足细胞焦亡的影响及其机制

Effect of gAd on podocyte pyroptosis under a high glucose condition and its mechanism

目的探讨脂联素球状结构域(gAd)对高糖环境下足细胞焦亡的影响及其可能的分子机制。方法采用正常浓度葡萄糖(5 mmol/L GS)和高浓度葡萄糖(25 mmol/L GS)培养足细胞,即对照组和高糖组,利用CCK-8法筛选出葡萄糖作用于足细胞的最佳时间。将足细胞分为正常(5 mmol/L GS)+PBS组、高糖(25 mmol/L GS)+PBS组、高糖(25 mmol/L GS)+5μg/ml gAd组,Western blot法检测细胞焦亡通路中Gasdermin D蛋白N端结构域(GSDMD-N)、GSDMD、Nod样受体蛋白3(NLRP3)、Caspase-1和pro-Caspase-1的表达。结果与对照组(5 mmol/L GS)相比,高糖组(25 mmol/L GS)细胞活性在高糖处理36 h出现显著降低(P<0.001),因此后续实验选取高糖处理细胞36 h。与正常+PBS组相比,高糖+PBS组GSDMD、Caspase-1和pro-Caspase-1表达水平均上调(P<0.05)。与高糖+PBS组相比,高糖+5μg/ml gAd组焦亡相关分子GSDMD、Caspase-1和pro-Caspase-1表达水平均下调(P<0.05)。结论gAd对高糖环境下的足细胞具有保护作用,可能与抑制NLRP3/Caspase-1/GSDMD焦亡通路有关。

Objective To investigate the effect of adiponectin globular domain(gAd)on high glucose(GS)-induced pyroptosis of podocytes and its underlying molecular mechanism.Methods Podocytes were cultured in 5 mmol/L GS and 25 mmol/L GS,namely control group and high glucose group,and the optimal duration of glucose exposure was determined by CCK-8 method.The podocytes were divided into three groups:normal glucose(5 mmol/L GS)+PBS group,high glucose(25 mmol/L GS)+PBS group,and high glucose(25 mmol/L GS)+5μg/ml gAd group.The expression of proteins related to the pyroptosis pathway in the three groups was analyzed using Western blot.Results After 36 h high glucose treatment,the cell activity in high glucose group was significantly decreased compared to control group(P<0.001).Therefore,podocytes treated with high glucose for 36 h were selected for subsequent experiments.The expression levels of GSDMD,Caspase-1,and pro-Caspase-1 were upregulated in high glucose+PBS group compared to normal glucose+PBS group(P<0.05).The expression levels of GSDMD,Caspase-1,and pro-Caspase-1 were downregulated in high glucose+5μg/ml gAd group compared to high glucose+PBS group(P<0.05).Conclusion The gAd can exhibit a protective effect on podocytes in a high glucose environment by inhibiting NLRP3/Caspase-1/GSDMD pyroptosis pathway.