实用医学杂志 ›› 2021, Vol. 37 ›› Issue (11): 1397-1402.doi: 10.3969/j.issn.1006⁃5725.2021.11.005

• 基础研究 • 上一篇    下一篇

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

马金兰,王青,牛浩宇   

  1. 1 青海大学附属医院眼科(西宁 810001);2 青海大学医学院眼科(西宁 810001)

  • 出版日期:2021-06-10 发布日期:2021-06-10
  • 基金资助:
    青海省科技计划项目(编号:2019⁃ZJ⁃7064)

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

MA Jinlan,WANG Qing,NIU Haoyu.    

  1. Department of Ophthalmology,Affiliated Hospital of Qinghai University,Xining 810001,China 

  • Online:2021-06-10 Published:2021-06-10

摘要:

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、20 μmol/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 20 μmol/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 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 ROS 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 ROS MDA 水平、凋亡率和 Bax 蛋白表达(P < 0.05),提高了 SOD 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。

关键词:

延龄草苷, 糖尿病视网膜病变, miR?1247?3p, 氧化应激, 凋亡

Abstract:

Objective To investigate the effect and mechanism of trillin on high glucose⁃induced retinal vascular endothelial cells(HRVEC)injury. Methods 5,10,and 20 μmol/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 20 μmol/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.

Key words:

trillin, diabetic retinopathy, miR?1247?3p, oxidative stress, apoptosis