长链非编码RNA生长停滞特异性转录本5在糖尿病肾病进展中作用机制研究  

Study on the mechanism of long non⁃coding RNAs growth arrest⁃specific transcript 5 in the progression of diabetic nephropathy

作　　者：李杨 徐曼 潘妙霞[1] 陈剑妹[1] 王玉川 蔡兴莉 LI Yang;XU Man;PAN Miao-xia;CHEN Jian-mei;WANG Yu-chuan;CAI Xing-li(Department of Nephrology and Rheumatology,Haikou Hospital,Xiangya Medical College,Central South University,Haikou 570208,China)

机构地区：[1]中南大学湘雅医学院附属海口医院肾病风湿科,海南海口570208

出　　处：《临床军医杂志》2023年第7期716-721,共6页Clinical Journal of Medical Officers

基　　金：海南省自然科学基金(819QN384);海南省自然科学基金(820QN422);海南省卫生健康行业科研项目(19A200125);海南省卫生健康行业科研项目(20A200321)。

摘　　要：目的探讨长链非编码RNA生长停滞特异性转录本5(LncRNA GAS5)在糖尿病肾病(DN)进展中的作用机制。方法选取10只健康5周龄大鼠为研究对象。将大鼠分为模型组与对照组,每组各5只。构建DN细胞模型,并分为LncRNA GAS5干扰组、LncRNA GAS5过表达组与阴性对照组。通过实时荧光定量聚合酶链式反应及蛋白质印迹法检测LncRNA GAS5、miR⁃135a⁃5p及沉默信息调节因子2相关酶1(SIRT1)的表达水平。采用双荧光素酶报告实验验证miR⁃135a⁃5p与LncRNA GAS5、SIRT1之间的相互作用。结果模型组肾组织、血清外泌体、人肾小球系膜细胞(HMC)及上清外泌体中LncRNA GAS5表达水平均低于对照组,差异均有统计学意义(P<0.05)。敲减或过表达LncRNA GAS5后,LncRNA GAS5过表达组HMC细胞中miR⁃135a⁃5p表达水平相应高于或低于阴性对照组,而SIRT1表达水平相应低于或高于阴性对照组,差异均有统计学意义(P<0.05)。感染miR⁃135a⁃5p或转染siSIRT1后,LncRNA GAS5过表达组对HMC细胞增殖的抑制作用及对α⁃平滑肌肌动蛋白、纤连蛋白、α(I)胶原表达的抑制作用均弱于阴性对照组,但差异无统计学意义(P>0.05)。结论LncRNA GAS5可通过miR⁃135a⁃5p/SIRT1通路调控DN的进展。Objective To investigate the mechanism of long non⁃coding RNAs growth arrest⁃specific transcript 5(LncRNA GAS5)in the progression of diabetic nephropathy(DN).Methods Ten healthy 5⁃week⁃old rats were selected as the study subjects.The rats were divided into model group and control group with 5 rats in each group.The DN cell model was constructed and divided into LncRNA GAS5 interference group,LncRNA GAS5 overexpression group and negative control group.The expression levels of LncRNA GAS5,miR⁃135a⁃5p and silent information regulator factor 2 related enzyme 1(SIRT1)were detected by real⁃time fluorescence quanti⁃tative polymerase chain reaction and western blot.Dual luciferase reporting experiments were used to verify the interaction between miR⁃135a⁃5p and LncRNA GAS5 and SIRT1.Results The expression levels of LncRNA GAS5 in kidney tissue,serum exosomes,hu⁃man mesangial cells(HMCs)and supernatant exosomes in model group were lower than those in control group,with statistical signifi⁃cance(P<0.05).After LncRNA GAS5 was knocked down or overexpressed,the expression level of miR⁃135a⁃5p in HMCs of Ln⁃cRNA GAS5 overexpression group was correspondingly higher or lower than that of negative control group,while the expression level of SIRT1 was lower or higher than that of negative control group,with statistical significance(P<0.05).After infection with miR⁃135a⁃5p or transfection with siSIRT1,the inhibition effect of LncRNA GAS5 overexpression group on HMC cell proliferation,andα⁃smooth muscle actin,fibronectin,α(I)collagen expression inhibition effect were weaker than that of negative control group,with no statistical significance(P>0.05).Conclusion Conclusion LncRNA GAS5 can regulate the progression of DN through miR⁃135a⁃5p/SIRT1 pathway.

关 键 词：糖尿病肾病 外泌体 长链非编码RNA生长停滞特异性转录本5 沉默信息调节因子2相关酶1 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]