Corilagin对OX-LDL诱导的HUVECs细胞损伤的保护作用及对MyD88信号通路表达的影响

Protective Effect of Corilagin on OX-LDL-Induced HUVECs Cell Damage and Its Impact on the Expression of the MyD88 Signaling Pathway

目的探讨不同浓度和时间的柯里拉京(Corilagin)处理对氧化性低密度脂蛋白(oxidative low-density lipoprotein,ox-LDL)诱导人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)损伤的保护作用,以及其对MyD88信号通路表达调控的影响。方法体外培养HUVECs复制ox-LDL诱导损伤模型,利用形态学及免疫学方法鉴定HUVECs细胞,MTT法确定复制ox-LDL损伤HUVECs模型的最佳条件,根据不同处理将细胞分为正常组(Normal)、模型组(Model)、Corilagin(3.125、6.25、12.5、25、50μmol/L)组、阳性对照(VE 10μmol/L、Simvastain 1μmol/L)组。观察Corilagin对ox-LDL诱导损伤的HUVECs保护作用;Western blot和RT-qPCR方法检测HUVECs细胞中MyD88、P65、TNF-α、MCP-1表达变化。结果经形态学及免疫学方法鉴定,所培养细胞为HUVECs;70 mg/L的ox-LDL刺激12 h是复制ox-LDL损伤HUVECs模型的最佳条件;MTT结果显示,与ox-LDL组比较,Corilagin组细胞活力明显升高(P<0.01);RT-qPCR和Western blot结果表明,与Normal组比,ox-LDL组MyD88、P65、TNF-α及MCP-1的mRNA和蛋白表达升高(P<0.01);与ox-LDL组比较,Corilagin组及阳性对照组的MyD88、P65、TNF-α及MCP-1的mRNA和蛋白表达降低(P<0.01),且Corilagin组呈现剂量依赖下调MyD88、P65、TNF-α及MCP-1的mRNA和蛋白表达。结论随着时间和浓度的增加,Corilagin能明显提高ox-LDL诱导损伤的HUVECs细胞活力,其保护作用与抑制MyD88信号通路有关。

Objective To investigate the protective effects of different concentrations and durations of corilagin treatment on oxidative low-density lipoprotein(ox-LDL)-induced damage in human umbilical vein endothelial cells(HUVECs),as well as its effects on the expression regulation of the MyD88 signaling pathway.Methods In vitro cultured HUVECs were used to replicate the ox-LDL-induced damage model.Morphological and immunological methods were employed to identify HUVECs cells.The MTT assay was used to determine the optimal conditions for replicating the ox-LDL-induced damage model in HUVECs.Cells were divided into different groups based on different treatments:normal group,model group,Corilagin(3.125,6.25,12.5,25,50μmol/L)group,and positive control(VE 10μmol/L,Simvastatin 1μmol/L)group.The protective effect of corilagin on ox-LDL-induced damage in HUVECs was observed.Western blot and RT-qPCR methods were used to detect the expression changes of MyD88,P65,TNF-α,and MCP-1 in HUVECs cells.Results The cultured cells were confirmed as HUVECs by using morphological and immunological methods.The best condition for replicating ox-LDL-induced damage to HUVECs was 12 hours of treatment with 70 mg/L of ox-LDL.MTT results showed that compared with the ox-LDL group,the corilagin group had a significantly higher cell viability(P<0.01).RT-qPCR and Western blot results showed that compared with the control group,the mRNA and protein expression of MyD88,P65,TNF-α,and MCP-1 were increased in the ox-LDL group(P<0.01).Compared with the ox-LDL group,the mRNA and protein expression of MyD88,P65,TNF-α,and MCP-1 were decreased(P<0.01)in the corilagin group and positive control group,and the corilagin group showed a dose-dependent downregulation of the mRNA and protein expression of MyD88,P65,TNF-α,and MCP-1.Conclusion With the increase of time and concentration,corilagin can significantly improve the cell viability of HUVECs induced by ox-LDL damage,and its protective effect is associated with the inhibition of the MyD88 signaling pathway.