血管生成素样蛋白8促进大鼠主动脉血管平滑肌细胞表型转化

Angiopoietin-like protein 8 promotes phenotypic transformation of rat aorta vascular smooth muscle cells

目的探讨血管生成素样蛋白8(ANGPTL8)对大鼠主动脉血管平滑肌细胞(VSMCs)表型转化的作用机制。方法ApoE-/-小鼠西方饮食12周构建动脉粥样硬化模型小鼠。免疫荧光染色检测ANGPTL8在动脉硬化斑块中的表达与定位;采用慢病毒转染VSMCs构建ANGPTL8过表达稳转细胞株;将细胞随机分为5组,对照(control)组、ANGPTL8过表达(LV-ANGPTL8)组、ox-LDL(50 mg/L)组、(ox-LDL+ANGPTL8过表达)组、[ANGPTL8过表达+ERK通路抑制剂(PD98059)]组。划痕实验观察细胞迁移;Western blot检测VSMCs中α-SMA、SM22α、OPN、PCNA、Ki67的蛋白表达。结果过表达ANGPTL8后促进了VSMCs中OPN、PCNA、Ki67的蛋白表达(P<0.05),抑制了α-SMA、SM22α的蛋白表达(P<0.05),促进了VSMCs的迁移能力(P<0.05);加入ERK通路抑制剂PD98059后OPN、PCNA、Ki67的蛋白表达减少(P<0.05),α-SMA、SM22α的蛋白表达增加(P<0.05),VSMCs的迁移能力降低(P<0.05)。结论ANGPTL8可能通过激活ERK通路促进VSMCs的表型转化。

Objective To explore the role and mechanism of angiopoietin-like protein 8(ANGPTL8)on phenotypic transformation of rat aorta vascular smooth muscle cells(VSMCs).Methods ApoE-/-male mice(8 weeks of age)were fed with a western diet for 12 weeks to build the atherosclerosis mouse model.The expression and localization of ANGPTL8 in atherosclerotic plaques were detected by immunofluorescence staining.Stable transfections of the ANGPTL8 over-expression vector were performed.VSMCs were randomly divided into 5 groups:control group,LV-ANGPTL8 group,ox-LDL(50 mg/L)group,(ox-LDL+LV-ANGPTL8)group and LV-ANGPTL8+PD98059 group.And cell migration was observed by wound-healing assay.The protein expressions ofα-SMA,SM22α,OPN,PCNA and Ki67 in VSMCs was detected by Western blot.Results ANGPTL8 over-expression promoted the expression of OPN,PCNA and Ki67 in VSMCs(P<0.05),decreased the expression ofα-SMA and SM22α(P<0.05)and enhanced migration ability of VSMCs(P<0.05).After adding ERK pathway inhibitor PD98059,the expression of OPN,PCNA and Ki67 decreased(P<0.05),α-SMA and SM22αexpression increased(P<0.05),and the migration capability of the VSMCs decreased(P<0.05).Conclusions ANGPTL8 maybe promotes phenotypic transformation of VSMCs by activating ERK pathway.