miR-1908抑制高糖诱导的人视网膜血管内皮细胞系HRECs凋亡

基础医学与临床 ›› 2020, Vol. 40 ›› Issue (8): 1090-1095.

miR-1908抑制高糖诱导的人视网膜血管内皮细胞系HRECs凋亡

周丽萍, 毛晓春^*, 李琴

襄阳市中心医院眼科, 湖北襄阳 441000

收稿日期:2019-06-13 修回日期:2019-11-04 出版日期:2020-08-05 发布日期:2020-07-29
通讯作者: ^*mlchunfen@163.com
基金资助:
武汉市卫生和计划生育委员会科研项目资助(WX15D02)

miR-1908 inhibits apoptosis of high glucose-induced human retinal vascular endothelial cell line HRECs

ZHOU Li-ping, MAO Xiao-chun^*, LI Qin

Department of Ophthalmology, Xiangyang Central Hospital, Xiangyang 441000, China

Received:2019-06-13 Revised:2019-11-04 Online:2020-08-05 Published:2020-07-29
Contact: ^*mlchunfen@163.com

摘要/Abstract

摘要： 目的探讨miR-1908对高糖诱导的人视网膜血管内皮细胞(HRECs)凋亡的影响及分子机制。方法培养HRECs细胞,分为对照组和高糖组,RT-qPCR检测细胞中miR-1908表达水平。转染miR-1908 模拟物(mimics)、整合素连接激酶(ILK)的小干扰RNA至HRECs细胞,qRT-PCR和Western blot分别检测miR-1908和ILK蛋白表达验证转染效率。使用高糖干预过表达miR-1908或ILK表达抑制的HRECs细胞,流式细胞仪检测凋亡率,Western blot检测B淋巴细胞瘤-2 (Bcl-2)和B淋巴细胞瘤-2相关蛋白(Bax)表达水平。双荧光素酶报告基因实验验证miR-1908和ILK之间关系。结果与对照组比,高糖组HRECs细胞中miR-1908的表达水平显著降低(P＜0.05)。过表达miR-1908或ILK表达可降低高糖诱导的HRECs细胞凋亡率(P＜0.05),上调Bcl-2蛋白表达(P＜0.05),下调Bax蛋白表达(P＜0.05)。miR-1908负调控ILK表达,ILK过表达逆转了miR-1908对HRECs细胞凋亡率、Bcl-2和Bax蛋白表达的影响。结论 miR-1908可能通过负调控ILK表达上调Bcl-2蛋白表达,下调Bax蛋白表达抑制HRECs细胞的凋亡。

关键词: 人视网膜血管内皮细胞, 高糖, miR-1908, 整合素连接激酶, 细胞凋亡

Abstract: Objective To investigate the effect of miR-1908 on apoptosis of human retinal vascular endothelial cells (HRECs) induced by high glucose concentration and its molecular mechanism. Methods HRECs cells were cultured and divided into normal group and high glucose group. The expression of miR-1908 was detected by RT-qPCR. miR-1908 mimics or small interfering RNA of ILK was transfected into HRECs cells, the expression miR-1908 and ILK protein was detected by RT-qPCR and Western blot in order to detect the transfection efficiency, respectively. Then high concentration of glucose was used to interfere with HRECs with miR-1908 over-expressed or ILK expression inhibited. Apoptosis rate was detected by flow cytometry, Bcl-2 and Bax protein expression was detected by Western blot. The dual luciferase reporter gene assay validated the relationship between miR-1908 and ILK. Results Compared with the control group, the expression level of miR-1908 in HRECs of high glucose group was significantly decreased (P＜0.05). miR-1908 over-expressed of or ILK expression decreased the apoptosis rate of HRECs induced by high concentration of glucose (P＜0.05), up-regulated Bcl-2 protein expression (P＜0.05), and down-regulated Bax protein expression (P＜0.05). miR-1908 negatively regulated ILK expression, and over-expression of ILK partially reversed the effect of miR-1908 on apoptosis rate, Bcl-2 and Bax protein expression in HRECs. Conclusions miR-1908 up-regulates the expression of Bcl-2 protein and down-regulates the expression of Bax protein to inhibit the apoptosis of HRECs via negatively regulating ILK expression.

Key words: retinal vascular endothelial cells, high glucose level, miR-1908, integrin-linked kinase, cell apoptosis

中图分类号:

R774.5

周丽萍, 毛晓春, 李琴. miR-1908抑制高糖诱导的人视网膜血管内皮细胞系HRECs凋亡[J]. 基础医学与临床, 2020, 40(8): 1090-1095.

ZHOU Li-ping, MAO Xiao-chun, LI Qin. miR-1908 inhibits apoptosis of high glucose-induced human retinal vascular endothelial cell line HRECs[J]. Basic & Clinical Medicine, 2020, 40(8): 1090-1095.

参考文献

[1] Wang SY, Andrews CA, Herman WH, et al. Incidence and risk factors for developing diabetic retinopathy among youths with type 1 or type 2 diabetes throughout the united states[J]. Ophthalmol, 2017, 124:424-430.
[2] Gai CL, Zhao JR, Chen XL, et al. Effects of high blood glucose fluctuation on DNA damage of diabetic rat retinal tissues[J]. Int Eye Sci, 2014, 14:992-995.
[3] 李疏凤, 胡健艳, 李婷婷, 等. 糖尿病视网膜病变神经损伤的病理改变及其相关分子机制的研究现状与进展[J]. 中华眼底病杂志, 2017, 33:312-315.
[4] 黄俊, 毛新帮. 微小RNA在糖尿病视网膜病变新生血管生成中的研究进展[J]. 中华实验眼科杂志, 2017, 35:478-480.
[5] 李化, 李彦, 周善璧. miRNA-142-5p在大鼠角膜新生血管中的作用及机制研究[J]. 第三军医大学学报, 2018, 40: 699-704.
[6] Zhang X, Di G, Dong M, et al. Epithelium-derived miR-204 inhibits corneal neovascularization[J]. Exp Eye Res, 2018, 167:122-127.
[7] 李青兰, 吕红彬, 苟文军, 等. 微小RNA芯片检测高糖环境下人视网膜微血管内皮细胞微小RNA的差异表达[J]. 中华眼科杂志, 2012, 48:604-609.
[8] 吕伯昌, 刘蓓, 刘静, 等. 基因沉默解离素-金属蛋白酶17(ADAM17)对高糖环境下人视网膜微血管内皮细胞通透性、增殖、迁移以及蛋白表达的影响[J]. 眼科新进展, 2017, 37:205-209.
[9] 刘晓瑞, 黄彬洋, 王岚, 等. 血糖波动与糖尿病并发症关系的研究进展[J]. 世界最新医学信息文摘, 2017, 5:78-79.
[10] 陈洁, 陈兴强, 符薇薇. 雷帕霉素对高糖诱导的大鼠肾系膜细胞增殖、凋亡和细胞周期的影响[J]. 中国免疫学杂志, 2017, 33:47-51.
[11] Ma Y, Feng J, Xing X, et al. miR-1908 over-expression inhibits proliferation, changing akt activity and p53 expression in hypoxic NSCLC Cells[J]. Oncol Res, 2016, 24:9-15.
[12] Chai Z, Fan H, Li Y, et al. miR-1908 as a novel prognosis marker of glioma via promoting malignant phenotype and modulating SPRY4/RAF1 axis[J]. Oncol Rep, 2017, 38:2717-2726.
[13] Cao MQ, You AB, Zhu XD, et al. miR-182-5p pro-motes hepatocellular carcinoma progression by repressing FOXO3a[J]. J Hematol Oncol, 2018, 11:12-21.
[14] Ding F, Zhang S, Gao S, et al. miR-137 functions as a tumor suppressor in pancreatic cancer by targeting mrgbp: mir-137 in pancreatic cancer[J]. J Cell Biochem, 2018, 119:4799-4807.
[15] Li Y, Zhang J, Yan H. Integrin-linked kinase inhibition attenuates permeability of the streptozotocin-induced diabetic rat retina[J]. Cell Biochem Biophys, 2013, 67:1467-1472.