基于JAK2/STAT3信号通路探讨黄芪-山茱萸对高糖诱导足细胞损伤的保护作用

Experimental Study on Protective Effects of Huangqi-Sanzhuyu(Astragalus Membranaceus-Cornus Officinalis)on High Glucose-Induced Podocyte Injury by Inhibiting JAK2/STAT3 Signal Pathway

目的:探究黄芪-山茱萸对高糖诱导肾足细胞损伤的保护作用及机制。方法:CCK8法检测黄芪-山茱萸(0、175、350、700、1400、2800 mg/L)对足细胞(MPC-5)活性的影响。30 mmol/L葡萄糖诱导MPC-5细胞损伤,350、700、1400 mg/L黄芪-山茱萸进行干预;ELISA测定MPC-5细胞IL-6、IL-1β水平,流式细胞术测定细胞凋亡,免疫荧光染色测定MPC-5肾病蛋白(nephrin)、足细胞素(podocin)表达,qRT-PCR测定MPC-5细胞nephrin、podocin、结蛋白(desmin)、Wilm瘤基因1(WT-1)基因表达,Western blot测定细胞Janus激酶2/信号转导和转录激活因子3(JAK2/STAT3)信号通路磷酸化表达。结果:2800 mg/L黄芪-山茱萸可显著抑制MPC-5细胞活性(P<0.01)。MPC-5细胞经高糖诱导后,IL-6、IL-1β水平升高,凋亡增加,nephrin、podocin、WT-1表达降低,desmin表达升高,JAK2/STAT3信号通路磷酸化表达增加(P<0.01)。350、700、1400 mg/L黄芪-山茱萸可抑制高糖诱导MPC-5细胞IL-6、IL-1β水平升高和凋亡,增强nephrin、podocin、WT-1表达,降低desmin表达,抑制JAK2/STAT3信号通路磷酸化表达(P<0.05)。结论:黄芪-山茱萸可减轻高糖诱导足细胞炎症和凋亡损伤,其机制与抑制JAK2/STAT3通路磷酸化有关。

Objective:To explore the improvement effect and mechanism of Astragalus membranaceus-Cornus officinalis(A-C)on podocyte injury induced by high glucose.Methods:CCK8 method was used to detect the activity of podocyte cells(MPC-5)cultured by A-C(0,175,350,700,1400,2800 mg/L).High glucose(30 mmol/L glucose)was used to induce MPC-5 cell damage,350,700,1400 mg/LA-C to intervene.ELISA was used to detect the levels of IL-6 and IL-1βof MPC-5,flow cytometry was used to detect apoptosis of;immunofluorescence staining was used to detect the expression of nephrin,podocin;qRT-PCR was used to detect the gene expression of nephrin,podocin,desmin,WT-1;Western blot was used to detect the phosphorylation of JAK2/STAT3 signal pathway.Results:2800 mg/L A-C significantly inhibited MPC-5 activity(P<0.01).The induction of high glucose increased levels of IL-6,IL-1βand apoptosis of MPC-5,reduced expression of nephrin,podocin,WT-1,increased desmin expression and phosphorylation of JAK2/STAT3 signal pathway(P<0.01).350,700,1400 mg/L A-C inhibited levels of IL-6,IL-1βand apoptosis of MPC-5 induced by high glucose,also increased expression of nephrin,podocin,WT-1,decreased desmin expression and phosphorylation of JAK2/STAT3 signal pathway(P<0.05).Conslusion:A-C alleviate inflammation and apoptosis injury of podocyte induced by high glucose,and its mechanism may be related to inhibiting JAK2/STAT3 pathway phosphorylation.