摘要
目的:通过构建小鼠足细胞损伤模型,探讨雷公藤甲素(TP)通过调控NLRP3/GSDMD/IL-1β通路对足细胞的保护作用。方法:培养小鼠足细胞MPC5,通过构建Cr(Ⅵ)足细胞损伤模型,分为正常组(Control组)、Cr(Ⅵ)组、Cr(Ⅵ)+TP组(TP组)。通过CCK-8法分别检测30~150μmol/L Cr(Ⅵ)作用于足细胞后对足细胞生存活力的影响和2~10 ng/ml TP作用于Cr(Ⅵ)染毒后的足细胞,以确定Cr(Ⅵ)和TP干预的最佳浓度和时间,Western blot法检测NLRP3、Caspase-1、GSDMD、IL-1β和IL-18焦亡相关蛋白的表达情况,细胞免疫荧光法检测足细胞WT-1蛋白表达情况。结果:CCK法结果表示,选取Cr(Ⅵ)浓度为150μmol/L,1 h干预细胞,TP浓度为2 ng/ml,24 h干预细胞。Western blot结果表明发现,与Control组相比,Cr(Ⅵ)组中NLRP3、Caspase-1、GSDMD、IL-1β和IL-18表达升高(P<0.05,P<0.01),TP组中NLRP3、Caspase-1、GSDMD、IL-1β和IL-18表达降低(P<0.05)。细胞免疫荧光结果显示,与Control组相比,Cr(Ⅵ)组中WT-1的阳性表达显著降低,经TP干预后表达升高。结论:TP能够改善Cr(Ⅵ)对足细胞的损伤作用并通过抑制NLRP3/GSDMD/IL-1β信号通路来实现的。
Objective:To explore the protective effect of triptolide(TP)on podocytes by regulating NLRP3/GSDMD/IL-1βpathway by establishing a mouse podocyte injury model.Methods:Mouse podocyte MPC5 was cultured and Cr(Ⅵ)podocyte injury model was established.The mice were divided into normal group(Control group),Cr(Ⅵ)group and Cr(Ⅵ)+TP group(TP group).The effect of 30~150μmol/L Cr(Ⅵ)on podocyte viability and the effect of 2~10 ng/ml TP on podocyte treated by Cr(Ⅵ)were detected by CCK-8 method to determine the best concentration and time of Cr(Ⅵ)and TP intervention.The expression of NLRP3,Caspase-1,GSDMD,IL-1βand IL-18 scorch related protein was detected by Western blot method,and the expression of WT-1 protein was detected by cellular immunofluorescence.Results:The results of CCK assay showed that the cells were treated with Cr(Ⅵ)at 150μmol/L for 1 h and TP at 2 ng/ml for 24 h.The results of Western blot showed that the expression of NLRP3,Caspase-1,GSDMD,IL-1βand IL-18 in Cr(Ⅵ)group was higher than that in Control group(P<0.05,P<0.01).The expression of NLRP3,Caspase-1,GSDMD,IL-1β,and IL-18 was decreased in the TP group(P<0.05).The results of cellular immunofluorescence showed that the positive expression of WT-1 in Cr(Ⅵ)group was significantly lower than that in Control group.The expression was elevated after the TP intervention.Conclusion:TP can improve the injury effect of Cr(Ⅵ)on podocytes by inhibiting NLRP3/GSDMD/IL-1βsignal pathway.
作者
魏晓菲
王晨丹
WEI Xiaofei;WANG Chendan(The Third Clinical College of Shanxi University of Traditional Chinese Medicine,Taiyuan 030024,China)
出处
《陕西中医》
CAS
2024年第9期1172-1175,1185,共5页
Shaanxi Journal of Traditional Chinese Medicine
基金
山西省科技厅自然科学研究面上项目(202103021224382)
中国医师协会循证医学专业委员会肾科学组“PRO·润”基金资助项目(KYS2021-03-02-10)。
