延龄草苷通过上调miR⁃1247⁃3p表达减轻高糖诱导的视网膜血管内皮细胞损伤

Trillin attenuates high glucose⁃induced retinal vascular endothelial cell injury by up⁃regulating expression of miR⁃1247⁃3p

目的探讨延龄草苷对高糖诱导的人视网膜血管内皮细胞(HRVEC)损伤的影响及作用机制。方法5、10、20μmol/L的延龄草苷作用于30 mmol/L葡萄糖诱导的HRVEC细胞,或30 mmol/L葡萄糖诱导转染miR⁃1247⁃3p模拟物的HRVEC细胞,或20μmol/L的延龄草苷作用于30 mmol/L葡萄糖诱导的转染miR⁃1247⁃3p抑制剂的HRVEC细胞,DCFH⁃DA法检测细胞中ROS水平,酶联免疫吸附法检测MDA和SOD水平,流式细胞仪检测细胞凋亡,Western blot法检测Bcl⁃2和Bax蛋白表达,RT⁃qPCR法检测miR⁃1247⁃3p表达。结果延龄草苷降低了高糖诱导的HRVEC中ROS和MDA水平、凋亡率及Bax蛋白表达(P<0.05),提高了SOD活性、Bcl⁃2蛋白表达及miR⁃1247⁃3p表达(P<0.05),且呈剂量依赖性。过表达miR⁃1247⁃3p降低了高糖诱导的HRVEC中ROS和MDA水平、凋亡率和Bax蛋白表达(P<0.05),提高了SOD活性和Bcl⁃2蛋白表达(P<0.05)。抑制miR⁃1247⁃3p逆转延龄草苷对高糖诱导的HRVEC细胞氧化应激和凋亡的影响(P<0.05)。结论延龄草苷可能通过上调miR⁃1247⁃3p表达抑制高糖诱导的HRVEC细胞氧化应激和凋亡,减轻细胞损伤。

Objective To investigate the effect and mechanism of trillin on high glucose⁃induced retinal vascular endothelial cells(HRVEC)injury.Methods 5,10,and 20μmol/L of trillin were applied to HRVEC cells induced by glucose of 30 mmol/L,or 30 mmol/L of glucose was induced HRVEC cells transfected with miR⁃1247⁃3p mimic,or 20μmol/L of trillin was applied to 30 mmol/L glucose⁃induced HRVEC cells transfected with miR⁃1247⁃3p inhibitor.DCFH⁃DA method was used to detect intracellular ROS level;enzyme⁃linked immunosor⁃bent assay was used to detect levels of MDA and SOD;flow cytometry was used to detect cell apoptosis;Western Blot method was used to detect the protein expression levels of Bcl⁃2 and Bax;RT⁃qPCR method was used to detect the expression level of miR⁃1247⁃3p.Results Trillin reduced the levels of ROS and MDA,apoptosis rate,and protein expression of Bax in glucose⁃induced HRVEC cells(P<0.05),while increased SOD activity and expressions of Bcl⁃2 protein and miR⁃1247⁃3p(P<0.05)in a dose⁃dependent pattern.Overexpression of miR⁃1247⁃3p reduced the levels of ROS and MDA,apoptosis rate,and protein expression of Bax in glucose⁃induced HRVEC cells(P<0.05),while increased SOD activity and expression of Bcl⁃2 protein(P<0.05).Inhibition of miR⁃1247⁃3p reversed the effect of trillin on oxidative stress and apoptosis of HRVEC cells induced by high glucose(P<0.05).Conclusions Trillin may inhibit high glucose⁃induced oxidative stress and apoptosis in HRVEC cells by up⁃regu⁃lating the expression of miR⁃1247⁃3p,alleviating cell damage.