小檗碱抑制JAK2/STAT3信号通路缓解高糖诱导的足细胞EMT和凋亡  

Berberine inhibits JAK2/STAT3 signaling pathway to alleviate high glucose induced podocyte EMT and apoptosis

作　　者：吴昊 杨琳 唐丽琴[1,2] 魏伟 Wu Hao;Yang Lin;Tang Liqin;Wei Wei(Institute of Clinical Pharmacology,Anhui Medical University,Key Laboratory of Anti-inflammatory Immunodrugs of Ministry of Education,Hefei 230032;Dept of Pharmacy,The First Affiliated Hospital of University of Science and Technology of China,Hefei 230001)

机构地区：[1]安徽医科大学临床药理研究所,抗炎免疫药物教育部重点实验室,合肥230032 [2]中国科学技术大学附属第一医院药剂科,合肥230001

出　　处：《安徽医科大学学报》2022年第8期1189-1194,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:82174031、81773955);安徽省省级教学研究项目(编号:2020jyxm2328)。

摘　　要：目的探讨小檗碱(BBR)对高糖(HG)环境诱导的足细胞上皮间质转化(EMT)和凋亡的调节作用及部分机制。方法将指数生长期的足细胞分为以下几组:正常组、高糖组、小檗碱(30、60和90μmol/L)给药组。流式细胞术检测足细胞凋亡情况;Transwell和划痕实验检测足细胞迁移能力;Western blot检测细胞凋亡蛋白Bim、足细胞EMT相关蛋白nephrin、α-平滑肌肌动蛋白(α-SMA)、波形蛋白(vimentin)和酪氨酸激酶2(JAK2)/信号转导子和转录激活子3(STAT3)通路相关蛋白的表达水平;激光共聚焦法检测EMT相关蛋白nephrin、α-SMA和vimentin在足细胞中的表达情况。结果流式细胞术结果表明BBR(30、60和90μmol/L)能够减少高糖环境下的足细胞凋亡;Transwell和划痕实验结果表明BBR(30、60和90μmol/L)能抑制高糖环境下的足细胞异常迁移;Western blot结果表明BBR(30、60和90μmol/L)可以降低高糖环境下足细胞中p-JAK2、p-STAT3、α-SMA、vimentin、Bim蛋白的表达,升高nephrin蛋白的表达;激光共聚焦结果表明BBR(30、60和90μmol/L)可降低足细胞的细胞质上α-SMA、vimentin的表达,升高足细胞核和细胞质上nephrin的表达。结论BBR可能通过抑制JAK2/STAT3信号通路缓解高糖诱导的足细胞EMT和凋亡。Objective To investigate the regulatory effects and partial mechanisms of berberine(BBR)on podocyte epithelial mesenchymal transformation(EMT)and apoptosis induced by high glucose(HG).Methods Podocytes at exponential growth stage were divided into normal group,high glucose group and berberine group(30,60 and 90μmol/L).The rate of podocyte apoptosis was detected by flow cytometry.Transwell and scratch assay were used to detect podocyte migration.Western blot was used to detect the expression of apoptosis protein Bim,podocyte EMT-associated proteins nephrin,α-smooth muscle actin(α-SMA),vimentin and tyrosine kinase 2(JAK2)/signal transductor and transcriptional activator 3(STAT3)pathway-related proteins.Laser confocal method was used to detect the expression of EMT-related proteins nephrin,α-SMA and vimentin in podocytes.Results Flow cytometry results showed that BBR(30,60 and 90μmol/L)could reduce the rate of podocyte apoptosis in high glucose environment.Transwell and scratch test results showed that BBR(30,60 and 90μmol/L)could inhibit abnormal podocytes migration in high glucose environment.Western blot results showed that BBR(30,60 and 90μmol/L)could decrease the protein expression of p-JAK2,p-STAT3,α-SMA,vimentin and Bim,and increase the protein expression of nephrin in podocytes under high glucose condition.Laser confocal results showed that the BBR(30,60,and 90μmol/L)could decrease the expression ofα-SMA and vimentin in podocyte cytoplasm,increase the expression of nephrin in podocyte nucleus and cytoplasm.Conclusion BBR may alleviate the EMT and apoptosis of podocytes induced by high glucose through JAK2/STAT3 signaling pathway.

关 键 词：小檗碱 糖尿病肾病 足细胞 上皮间质转化 凋亡 

分 类 号：R587.24[医药卫生—内分泌]