杜仲绿原酸对LPS诱导的视网膜内皮细胞凋亡和免疫反应的调控作用

目的:糖尿病性视网膜病变是糖尿病最常见的并发症之一。本文旨在研究杜仲绿原酸(CA)对脂多糖(LPS)诱导的视网膜内皮细胞(HRECs)凋亡和炎症反应的影响。方法:CCK-8检测细胞增殖;流式细胞术分析细胞凋亡; ELISA检测炎症因子IL-6、TNF-α和IL-10水平。蛋白印迹检测Ki67、Bcl-2、Caspase-3、Bax、NF-κB P65和P-NF-κB P65蛋白水平。结果:低浓度(<50μmol/L)的杜仲绿原酸对HRECs细胞活力无明显影响,高浓度(>50μmol/L)的杜仲绿原酸会减低HRECs细胞活力。与对照组相比,LPS组细胞增殖明显降低,细胞凋亡明显上升(P<0. 05)。与LPS组相比,20和50μmol/L的杜仲绿原酸组细胞增殖升高,细胞凋亡下降(P<0. 05)。LPS组Ki67和Bcl-2表达明显低于对照组,Caspase-3和Bax表达明显高于对照组(P<0. 05)。与LPS组相比,20和50μmol/L的杜仲绿原酸组Ki67和Bcl-2表达明显升高; Caspase-3和Bax表达明显降低(P<0. 05)。而且,LPS组IL-6和TNF-α水平高于对照组,IL-10水平低于对照组(P<0. 05)。与LPS组相比,20和50μmol/L的杜仲绿原酸组IL-6和TNF-α水平下降,IL-10水平上升(P<0. 05)。另外,LPS组p-P65/P65比值高于对照组(P<0. 05)。杜仲绿原酸(10,20,50μmol/L)组p-P65/P65比值低于LPS组(P<0. 05)。结论:杜仲绿原酸通过抑制NF-κB P65活化减弱LPS诱导的HRECs细胞凋亡和炎症反应的增强。

Chlorogenic acid regulates lipopolysaccharide-induced apoptosis and inflammatory response of human retinal vascular endothelial cells

Objective:Diabetic retinopathy is one of the most common complications of diabetes.This study aims to explore the effect of chlorogenic acid(CA)on lipopolysaccharide(LPS)-induced apoptosis and inflammatory response of human retinal vascular endothelial cells(HRECs).Methods:Cell proliferation was tested by CCK-8.Cell apoptosis was detected by flow cytometry.The levels of IL-6,TNF-αand IL-10 were measured by ELISA.The protein levels of Ki67,Bcl-2,Caspase-3,Bax,NF-κB P65 and P-NF-κB P65 were detected by Western blot.Results:The low concentration(<50μmol/L)of CA had no effect on cell viability of HRECs.The cell viability of HRECs was decreased by high concentration( >50μmol/L)of CA.Compared with control group,the proliferation in LPS group was reduced with enhancive apoptosis(P<0.05).Compared with LPS group,the proliferation in CA(20,50μmol/L)group was increased with attenuated apoptosis(P<0.05).The protein levels of Ki67 and Bcl-2 in LPS group were lower than control group(P<0.05).And the expression of Caspase-3 and Bax in LPS group was higher than control group(P<0.05).Compared with LPS group,the protein levels of Ki67 and Bcl-2 in CA(20,50μmol/L)group were elevated with decreased expression of Caspase-3 and Bax(P<0.05).Moreover,the levels of IL-6 and TNF-αin LPS group were higher than control group(P<0.05).And the levels of IL-10 in LPS group were lower than control group(P<0.05).Compared with LPS group,the levels of IL-6 and TNF-αin CA(20,50μmol/L)group were reduced with enhanced levels of IL-10(P<0.05).In addition,the rate of p-P65/P65 in LPS group was higher than control group(P<0.05).The rate of p-P65/P65 in CA(10,20,50μmol/L)group was lower than LPS group(P<0.05).Conclusion:Chlorogenic acid alleviates LPS-induced enhancive apoptosis and inflammatory response of HRECs via inhibiting activation of NF-κB P65.