LncRNA NORAD对高糖诱导的人肾小管上皮细胞炎症因子和纤维化因子表达的影响及机制研究  

The effect and mechanism of LncRNA NORAD on the expression of inflammatory factors and fibrotic factors in human renal tubular epithelial cells induced by high glucose

作　　者：卢迪[1] 黎瑶[1] 李婷 邱平[1] Lu Di;Li Yao;Li Ting;Qiu Ping(Department of Endocrinology and Metabolic Diseases,The First Affiliated Hospital of Chengdu Medical College,Chengdu 610500,China)

机构地区：[1]四川成都医学院第一附属医院内分泌与代谢病科,成都610500

出　　处：《广西医科大学学报》2021年第9期1711-1716,共6页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81602821);四川省卫生和计划生育委员会普及应用项目资助(No.18PJ346)。

摘　　要：目的:探讨LncRNA NORAD对高糖诱导的人肾小管上皮细胞中炎性和纤维化因子表达的影响及分子机制。方法:将人肾小管上皮细胞HK-2分为空白对照(NC)组、渗透压对照(OSM)组、高糖处理(HG)组、si-NC+HG组、si-LncRNA NORAD+HG组、miR-NC+HG组、miR-367-3p+HG组、anti-miR-NC+si-LncRNA NORAD+HG组、anti-miR-367-3p+si-LncRNA NORAD+HG组。实时荧光定量PCR(q PCR)检测LncRNA NORAD和miR-367-3p的表达水平;蛋白质印迹(Western blotting)法检测α-SMA、Fn、COL1a1、COL3a1蛋白表达;酶联免疫吸附试验(ELISA)检测TGF-β1、IL-1β水平;双荧光素酶报告实验检测LncRNA NORAD和miR-367-3p的靶向关系。结果:高糖诱导的人肾小管上皮细胞中LncRNA NORAD表达水平升高,miR-367-3p表达水平降低(P<0.05);α-SMA、Fn、COL1a1、COL3a1蛋白的表达水平升高,TGF-β1、IL-1β水平升高(P<0.05)。低表达LncRNA NORAD或过表达miR-367-3p,高糖诱导的人肾小管上皮细胞中α-SMA、Fn、COL1a1、COL3a1蛋白的表达水平降低,TGF-β1、IL-1β水平降低(P<0.05)。LncRNA NORAD靶向调控miR-367-3p;低表达miR-367-3p可以逆转LncRNA NORAD低表达对高糖诱导的肾小管上皮细胞炎性和纤维化因子表达的抑制作用。结论:抑制LncRNA NORAD表达可能通过靶向上调miR-367-3p抑制高糖诱导的人肾小管上皮细胞中炎性和纤维化因子表达。Objective: To explore the effect and mechanism of LncRNA NORAD on the expression of inflammatory and fibrotic factors in human renal tubular epithelial cells induced by high glucose(HG). Methods: Human renal tubular epithelial cells HK-2 were divided into blank control(NC) group, osmotic pressure control(OSM)group, HG group, si-NC+HG group, si-LncRNA NORAD+HG group, miR-NC+HG group, miR-367-3 p+HG group, anti-miR-NC+si-LncRNA NORAD+HG group, anti-miR-367-3 p+si-LncRNA NORAD+HG group. Realtime fluorescent quantitative PCR(qPCR) was used to detect the expression levels of LncRNA NORAD and miR-367-3 p. Western blotting was used to detect the expression of α-SMA, Fn, COL1 a1, and COL3 a1 protein. Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of TGF-β1 and IL-1β. Dual luciferase reporter assay was used to confirm the targeting relationship between LncRNA NORAD and miR-367-3 p. Results: The expression levels of LncRNA NORAD, α-SMA, Fn, COL1 a1 and COL3 a1 protein as well as the TGF-β1 and IL-1β levels in human renal tubular epithelial cells induced by HG were increased, while the expression level of miR-367-3 p was decreased(P<0.05). Low-expression of LncRNA NORAD or over-expression of miR367-3 p led to dramatic reductions in the expressions of α-SMA, Fn, COL1 a1 and COL3 a1 protein and the levels of TGF-β1 and IL-1β(P<0.05). LncRNA NORAD targeted and regulated miR-367-3 p. Low-expression of miR-367-3 p reversed the inhibitory effects of LncRNA NORAD low-expressionon the expression of inflammatory and fibrotic factors in renal tubular epithelial cells induced by HG. Conclusion: Inhibiting LncRNA NORAD expression may inhibit the expression of inflammatory and fibrotic factors in human renal tubular epithelial cells induced by HG by up-regulation of miR-367-3 p.

关 键 词：LncRNA NORAD miR-367-3p 肾小管上皮细胞 炎症因子 纤维化 

分 类 号：R587.2[医药卫生—内分泌]