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第41卷第12期 南京医科大学学报(自然科学版)
2021年12月 Journal of Nanjing Medical University(Natural Sciences) ·1741 ·
·基础医学·
长链非编码RNA ENSMUST00000127391 对小鼠肾小球系膜细
胞增殖和纤维化的影响
李兰兰,陈 欣,张恒璐,王 敏 ,陆卫平 *
*
南京医科大学附属淮安第一医院内分泌科,江苏 淮安 223300
[摘 要] 目的:探究长链非编码RNA ENSMUST00000127391(lncRNA 127391)在高糖培养的小鼠肾小球系膜细胞(mice me⁃
sangial cell,MMC)中的表达及功能。方法:在成功构建重组质粒 pCDH⁃lncRNA 127391 的基础上,将重组质粒 pCDH⁃lncRNA
127391和对照质粒pCDH分别转染MMC细胞,通过RT⁃qPCR 法检测lncRNA 127391表达水平。蛋白质免疫印迹法检测增殖
和纤维化相关蛋白水平;CCK8法检测lncRNA 127391对MMC细胞增殖的影响;流式细胞术检测对细胞凋亡的影响。结果:过
表达lncRNA 127391的MMC细胞构建成功。CCK8结果显示,lncRNA 127391表达增加后,MMC细胞增殖速度减慢;蛋白质免
疫印迹结果表明过表达lncRNA 127391后,MMC细胞中细胞周期蛋白D1(Cyclin D1)、增殖细胞核抗原(proliferating cell nucle⁃
ar antigen,PCNA)、纤维连接蛋白(FN)和Ⅰ型胶原(Col⁃1)蛋白表达水平均下降;流式分析结果显示,过表达lncRNA127391后,
MMC细胞凋亡数增加。两两比较,差异均有统计学意义(P < 0.05)。结论:lncRNA 127391可以抑制高糖培养的MMC细胞增
殖和纤维化。
[关键词] 长链非编码RNA;糖尿病肾病;增殖;纤维化
[中图分类号] R329.26 [文献标志码] A [文章编号] 1007⁃4368(2021)12⁃1741⁃06
doi:10.7655/NYDXBNS20211205
The effects of long non⁃coding RNA ENSMUST00000127391 on proliferation and fibrosis
of mice mesangial cells
*
LI Lanlan,CHEN Xin,ZHANG Henglu,WANG Min ,LU Weiping *
Department of Endocrinology and Metabolism,the Affiliated Huai’an NO.1 People’s Hospital of Nanjing Medical
University,Huai’an 223300,China
[Abstract] Objective:This study aims to investigate the expression and function of long non⁃coding RNA ENSMUST00000127391
(lncRNA 127391)in mice mesangial cell(MMC)cultured with high glucose. Methods:The recombinant plasmid pCDH ⁃ lncRNA
127391 was successfully constructed in our group. In this study,the recombinant plasmid pCDH⁃lncRNA 127391 and control plasmid
pCDH were transfected into MMC cells respectively,and the expression of lncRNA 127391 was detected by RT⁃qPCR. The fibrosis
markers of fibronectin(FN)and type Ⅰ collagen(Col⁃1)and proliferation markers of Cyclin D1 and proliferating cell nuclear antigen
(PCNA)were detected by Western blot. CCK8 assay was used to detect the proliferation of MMC. Flow cytometry was used to study the
effect of lncRNA 127391 on the apoptosis of MMC. Results:Overexpression of lncRNA 127391 in MMC cells was successfully
constructed. CCK8 showed that the overexpression of lncRNA 127391 decreased the proliferation rate of MMC cells. And the
expression of Cyclin D1,proliferating cell nuclear antigen PCNA,FN and Col ⁃ 1 in MMC cells decreased after overexpression of
lncRNA 127391. Flow cytometry showed that the overexpression of lncRNA127391 increased the number of apoptotic MMC cells. The
differences were statistically significant(P < 0.05). Conclusion:lncRNA 127391 can inhibit the proliferation and fibrosis of MMC
cells cultured with high glucose.
[Key words] long non⁃coding RNA;diabetic nephropathy;proliferation;fibrosis
[J Nanjing Med Univ,2021,41(12):1741⁃1746]
[基金项目] 国家自然科学青年基金(81800644);江苏省卫生健康委员会课题(H201458);淮安市533英才工程科研项目资助
计划(HAA201744)
∗
通信作者(Corresponding author),E⁃mail:736537823@qq.com;hayylwp@njmu.edu.cn