miR-155在高糖诱导足细胞损伤的表达及其与足细胞损伤相关蛋白表达的相关性

Expression of miR-155 in high glucose induced podocyte injury and its correlation with podocyte injury related protein expression

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.