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研究
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南京医科大学附属淮安第一医院内分泌科

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] 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)


The role of long non-coding RNA ENSMUST00000127391 in proliferation and fibrosis of mice mesangial cells
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Department of Endocrinology and Metabolism,the Affiliated Huai’an NO.1 People’s Hospital of Nanjing Medical University,Huai’an 223300

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    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 mesangial cell,MMC)中的表达及功能。方法:前期本课题组已成功构建重组质粒pCDH-lncRNA 127391,本研究将重组质粒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 nuclear antigen,PCNA)、纤维连接蛋白(FN)和I型胶原 (Col-1) 蛋白水平表达均下降;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。

    Abstract:

    Objective: To investigate the expression and function of long non?coding RNA ENSMUST00000127391 (lncRNA 127391) in mouse Mesangial cells (mice mesangial cell,MMC) cultured with high glucose.SMethods: 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.SThe Fibrosis markers of FN and Col-1 and Proliferation markers of Cyclin D1 and 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.SResults: Overexpression of lncRNA 127391 in MMC cells was successfully constructed.SCCK8 showed that the overexpression of lncRNA 127391 decreased the proliferation rate of MMC cells , and the expression of cyclin D1 (Cyclin D1), proliferating cell nuclear antigen ((proliferating cell nuclear antigen,PCNA), fibronectin (FN) and type I collagen (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.SThe differences were statistically significant (p<0.05).SConclusion: lncRNA 127391 can inhibit the proliferation and fibrosis of MMC cells cultured with high glucose.

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  • 收稿日期:2021-04-04
  • 最后修改日期:2021-07-12
  • 录用日期:2021-12-23
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