摘要
目的探讨高糖(HG)诱导足细胞损伤中miR-155的表达变化及miR-155与足细胞损伤相关蛋白表达的关系。方法体外培养人肾小球足细胞(AB8/13),用30 mmol/L的D-葡萄糖溶液刺激AB8/1348 h作为HG组,同时以终浓度为5 mmol/L的D-葡萄糖溶液处理细胞48 h作为正常对照(NC)组。用CCK8法检测细胞增殖活性。实时荧光定量聚合酶链反应(qRT-PCR)检测nephrin、F-actin、synaptopodin、p-cadherin的mRNA和miR-155的表达。Western blot法检测nephrin、F-actin、synaptopodin、p-cadherin蛋白的表达。鬼笔环肽染色观察细胞骨架变化。细胞免疫荧光检测蛋白nephrin、synaptopodin。结果HG可抑制足细胞增殖,降低细胞存活率(P<0.01)。HG作用的Ab8/13中的miR-155表达水平升高,nephrin、F-actin、synaptopodin、p-cadherin的mRNA与蛋白表达水平降低(P<0.05)。HG会导致足细胞骨架紊乱。结论HG作用的人足细胞中miR-155表达升高,miR-155表达与HG诱导的足细胞损伤有关,可作为足细胞损伤诊断和治疗的靶点。
Objective To investigate the changes of expression of miR-155 in podocyte injury induced by high glucose(HG)and its relationship with podocyte injury related protein expression.Methods Human glomerular podocytes(AB8/13)were cultured in vitro,AB8/13 stimulated with 30 mmol/L D-glucose solution for 48 h were selected as HG group,and cells treated with a final concentration of 5 mmol/L D-glucose solution for 48 h as normal control(NC)group.Cell proliferation activity was detected by CCK8.Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect nephrin,F-actin,synaptopodin,and the mRNA and miR-155 expressions of p-cadherin.Western blot was used to detect the protein expressions of nephrin,F-actin,synaptopodin and p-cadherin.Cytoskeleton changes were observed by phalloidin staining,and cellular immunofluorescence was used to detect protein nephrin and synaptopodin.Results HG could inhibit podocyte proliferation and decrease cell survival rate(P<0.01).The expression level of miR-155 in Ab8/13 induced by HG increased,while the mRNA and protein expression levels of nephrin,F-actin,synaptophodin,and p-cadherin decreased(P<0.05),and HG could cause disorder of foot cytoskeleton.Conclusion The expression of miR-155 in human podocytes induced by HG elevates,and miR-155 expression is associated with HG induced podocyte injury,which can be served as a target for the diagnosis and treatment of podocyte injury.
作者
王蓉
林栩
吴昱升
唐志明
王晨
韦美理
黄慕源
张洁
徐璐瑶
WANG Rong;LIN Xu;WU Yusheng;TANG Zhiming;WANG Chen;WEI Meili;HUANG Muyuan;ZHANG Jie;XU Luyao(Department of Nephrology,Affiliated Hospital of Youjiang Medical University for Nationalities,Baise 533000,Guangxi,China;Guangxi Key Laboratory of Basic Guarantee for Medical Research of Immune-related Diseases,Baise 533000,Guangxi,China)
出处
《右江医学》
2023年第4期300-306,共7页
Chinese Youjiang Medical Journal
基金
国家自然科学基金(81860131,82060133)
广西自然科学基金(2019GXNSFAA245004)
右江民族医学院研究生创新计划项目(YXCXJH2022001)。