阿魏酸改善高糖诱导的视网膜色素上皮细胞损伤的研究

Study on the improvement of high glucose-induced damage to retinal pigment epithelial cells by ferulic acid

目的 通过观察阿魏酸(FA)对高糖(HG)诱导视网膜色素上皮细胞(ARPE-19)损伤的影响,探讨FA对糖尿病视网膜病变的作用机制。方法 采用HG处理ARPE-19构建体外糖尿病视网膜色素上皮细胞损伤模型,FA溶解在二甲基亚砜(DMSO)中,分为Control组(5 mmol/L葡萄糖)、DMSO组(0.1%DMSO+5 mmol/L葡萄糖)、HG组(30 mmol/L葡萄糖)、HG+FA组(30 mmol/L葡萄糖+50μmol/L)。通过CCK-8检测细胞的增殖活性;采用活性氧(ROS)检测试剂盒检测各组ROS水平;免疫细胞荧光和Western blot检测Toll样受体4(TLR4)、髓样分化因子(MyD88)、核因子κB(NF-κB)的表达水平。结果 CCK-8检测结果表明,与Control组相比,HG可诱导ARPE-19细胞的增殖活性降低,而FA可改善高糖对细胞增殖的影响(P<0.05)。ROS检测结果表明,与Control组相比,HG组可诱导ARPE-19细胞ROS荧光强度升高,FA可显著降低高糖诱导ARPE-19细胞的ROS荧光强度(P<0.05)。免疫细胞荧光和Western blot实验表明,HG组可诱导ARPE-19细胞TLR4、MyD88、NF-κB表达升高,FA可显著降低它们的表达(P<0.05)。结论 FA通过降低ROS产生和下调TLR4、MyD88、NF-κB表达改善高糖诱导的视网膜色素上皮细胞损伤。

Objective To observe the effect of ferulic acid(FA)on the injury of retinal pigment epithelial cells(ARPE-19)induced by high glucose(HG),and to explore the mechanism of FA on diabetes retinopathy.Methods The model of diabetic retinopathy in vitro was constructed by culturing ARPE-19 with HG,and FA was dissolved in dimethyl sulfoxide(DMSO).They were divided into Control group(5 mmol/L glucose),DMSO group(0.1%DMSO+5 mmol/L glucose),HG group(30 mmol/L glucose)and HG+FA group(30 mmol/L glucose+50μmol/L).The proliferative activity of the cells was detected by CCK-8 assay.ROS levels of ARPE-19 cells were detected by reactive oxygen species(ROS)detection kit.The expression levels of TLR4,MyD88 and NF-κB were detected by immunofluorescence and Western blotting.Results Compared with the control group,HG decreased the proliferation activity of ARPE-19 cells,while FA ameliorated the effect of HG on the proliferation of cells(P<0.05).Compared with the control group,the ROS fluorescence intensity of ARPE-19 cells induced by HG increased,and the FA significantly decreased the ROS fluorescence intensity of ARPE-19 cells induced by HG(P<0.05).Immunofluorescence and Western blot experiments showed that the expression of TLR4,MyD88 and NF-κB in ARPE-19 cells increased in HG group,while they significantly decreased in FA group(P<0.05).Conclusion FA can improve the damage of retinal pigment epithelial cells induced by HG by decreasing ROS production and down-regulating the expression of TLR4,MyD88 and NF-κB.