lncRNA SNHG6对高糖诱导的人视网膜微血管内皮细胞损伤的影响  

Effect of lncRNA SNHG6 on high glucose-induced human retinal microvascular endothelial cell injury

作　　者：吴海星 周金红[2] 吴田莉 章沐曦[2] 李小义 张学东[3] Wu Haixing;Zhou Jinhong;Wu Tianli;Zhang Muxi;Li Xiaoyi;Zhang Xuedong(Department of Ophthalmology,Chenjiaqiao Hospital of Shapingba District,Chongqing 400000,China;Department of Ophthalmology,Suining First People's Hospital,Suining 629000,Sichuan Province,China;Department of Ophthalmology,the First Affiliated Hospital of Chongqing Medical University,Chongqing 400042,China)

机构地区：[1]中国重庆市沙坪坝区陈家桥医院眼科,400000 [2]中国四川省遂宁市第一人民医院眼科,629000 [3]重庆医科大学附属第一医院眼科,中国重庆市400042

出　　处：《国际眼科杂志》2024年第11期1715-1720,共6页International Eye Science

摘　　要：目的:探讨lncRNA SNHG6对高糖诱导的人视网膜微血管内皮细胞(hRMECs)损伤的影响及其可能的作用机制。方法:采用高糖诱导hRMECs建立细胞损伤模型。高糖(HG)组将hRMECs培养在浓度为25 mmol/L D-葡萄糖的DMEM培养基24 h;正常(NG)组将hRMECs培养在浓度为5.5 mmol/L D-葡萄糖的DMEM培养基;按照实验设计分别将si-NC、si-SNHG6、si-SNHG6和anti-miR-NC、si-SNHG6和anti-miR-186-5p转染至hRMECs,随后25 mmol/L D-葡萄糖孵育24 h,分别记为HG+si-NC组、HG+si-SNHG6组、HG+si-SNHG6+anti-miR-NC组、HG+si-SNHG6+anti-miR-186-5p组。qRT-PCR法检测lncRNA SNHG6、miR-186-5p的表达量;双荧光素酶报告实验检测lncRNA SNHG6与miR-186-5p的靶向关系;MTT法与流式细胞术分别检测细胞增殖及凋亡;ELISA法检测IL-1β、TNF-α、IL-8、IL-10的水平;采用试剂盒检测SOD的活性和MDA的水平;Western blot检测cleaved-caspase3、Bax、Bcl-2蛋白表达量。结果:HG组与NG组比较lncRNA SNHG6表达升高,miR-186-5p表达降低(均P<0.05);lncRNA SNHG6可靶向结合miR-186-5p。转染si-SNHG6后细胞增殖抑制率、凋亡率、cleaved-caspase3、Bax蛋白水平,IL-1β、TNF-α、IL-8含量以及MDA活性降低(P<0.05),而Bcl-2蛋白、IL-10含量以及SOD的活性升高(P<0.05)。共转染si-SNHG6和anti-miR-186-5p后,细胞增殖抑制率、凋亡率、cleaved-caspase3、Bax、IL-1β、TNF-α、IL-8以及MDA升高(P<0.05),而Bcl-2蛋白、IL-10和SOD降低(P<0.05)。结论:干扰lncRNA SNHG6表达可通过促进miR-186-5p表达而抑制高糖诱导的hRMECs凋亡、炎症反应及氧化应激。AIM:To explore the effect of lncRNA SNHG6 on injury of human retinal microvascular endothelial cells(hRMECs)induced by high glucose and its possible mechanism.METHODS:The D-glucose-induced hRMECs were used to establish normal glucose(NG)and high glucose(HG)cell injured model.In the HG group,the hRMECs were cultured in DMEM medium at a concentration of 25 mmol/L D-glucose for 24 h,while in the NC group,they were cultured in DMEM medium at a concentration of 5.5 mmol/L D-glucose;according to experimental design,si-NC,si-SNHG6,si-SNHG6 and anti-miR-NC and si-SNHG6 and anti-miR-186-5p were transfected into hRMECs,and then incubated at a concentration of 25 mmol/L D-glucose for 24 h,with HG+si-NC group,HG+si-SNHG6 group,HG+si-SNHG6+anti-miR-NC group and HG+si-SNHG6+anti-miR-186-5p group marked,respectively.The quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression of lncRNA SNHG6 and miR-186-5p;dual-luciferase reporter assay was used to detect the targeting relationship;MTT assay and flow cytometry were used to detect the cell proliferation and apoptosis,respectively;enzyme linked immunosorbent assay(ELISA)was used to detect the levels of IL-1β,TNF-α,IL-8,IL-10;testing kits were used to detect activity of SOD and level of MDA;the Western blot was used to detect the protein expression of cleaved-caspase3,Bax and Bcl-2.RESULTS:The lncRNA SNHG6 expression increased in the HG group,while miR-186-5p expression decreased(both P<0.05).There was target binding of lncRNA SNHG6 with miR-186-5p.After the transfection of si-SNHG6,cell inhibition rate,apoptosis rate,cleaved-caspase3,Bax protein levels,IL-1β,TNF-α,IL-8 contents,and MDA activity were decreased(P<0.05),while Bcl-2 protein,IL-10 contents,and SOD activity were increased(P<0.05).Co-transfection of si-SNHG6 and anti-miR-186-5p increased cell proliferation inhibition rate,apoptosis rate,cleaved-caspase3,Bax,IL-1β,TNF-α,IL-8,and MDA(P<0.05),but decreased Bcl-2,IL-10 and SOD(P<0.05).CONCLUSION:Interfering with lncRNA SNHG6 could inh

关 键 词：人视网膜微血管内皮细胞 lncRNA SNHG6 miR-186-5p 细胞增殖 凋亡 炎症 氧化应激 

分 类 号：R774.1[医药卫生—眼科] R587.2[医药卫生—临床医学]