[1]易红,彭睿,孙艳,等.lncRNA-Gm4419对高糖培养的肾小球系膜细胞增殖和纤维化的影响[J].第三军医大学学报,2017,39(03):216-222.
 Yi Hong,Peng Rui,Sun Yan,et al.Effect of lncRNA-Gm4419 on proliferation and fibrosis in high glucose-induced glomerulus mesangial cells[J].J Third Mil Med Univ,2017,39(03):216-222.
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lncRNA-Gm4419对高糖培养的肾小球系膜细胞增殖和纤维化的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
39卷
期数:
2017年第03期
页码:
216-222
栏目:
基础医学
出版日期:
2017-02-15

文章信息/Info

Title:
Effect of lncRNA-Gm4419 on proliferation and fibrosis in high glucose-induced glomerulus mesangial cells
作者:
易红彭睿孙艳罗勇军彭惠民李爱玲张政
重庆医科大学基础医学院:细胞生物学及遗传学教研室,生物信息学教研室,实验教学中心;第三军医大学高原军事医学系军事医学地理学教研室
Author(s):
Yi Hong Peng Rui Sun Yan Luo Yongjun Peng Huimin Li Ailing Zhang Zheng 

Department of Cell Biology and Genetics, Department of Bioinformatics, 4Experimental Teaching Center, College of Basic Medical Sciences, Chongqing Medical University, Chongqing, 400016; Department of Military Medical Geography, College of High Altitude Military Medicine, Third Military Medical University, Chongqing, 400038, China

关键词:
长链非编码RNAGm4419肾小球系膜细胞增殖纤维化
Keywords:
long non-coding RNA Gm4419 glomerular mesangial cells proliferation fibrosis
分类号:
R322.61; R329.28; R394.2
文献标志码:
A
摘要:
目的      探讨长链非编码Gm4419对高糖培养的小鼠肾小球系膜细胞增殖和纤维化的影响。方法      荧光定量PCR检测Gm4419在糖尿病肾病肾脏组织及高糖培养的肾小球系膜细胞中的表达水平。设计合成Gm4419 siRNA,同时构建pcDNA3.1(+)-Gm4419过表达质粒,并将Gm4419 siRNA和pcDNA3.1(+)-Gm4419过表达质粒分别转染至高低糖培养的肾小球系膜细胞,EdU法检测转染后细胞的增殖能力;Western blot检测肾脏纤维标记蛋白的表达水平情况。结果      Gm4419在糖尿病肾病肾脏组织及高糖培养的肾小球系膜细胞中的表达水平较正常组及低糖组显著增高(P<0.01)。荧光定量PCR筛选出一条最佳干扰Gm4419的siRNA,转染高糖组后,与高糖mock组或对照组比较,高糖Gm4419 knockdown组细胞增殖能力减弱、纤维化标记蛋白表达减少(P<0.05)。此外,将酶切和测序结果证实构建成功的pcDNA3.1(+)-Gm4419过表达质粒转染至低糖组细胞,与低糖mock组或对照组相比,低糖Gm4419过表达组细胞增殖能力增强、纤维化标记蛋白表达水平增高(P<0.05)。 结论      lncRNA-Gm4419参与糖尿病肾小球系膜增生和纤维化的调节。
Abstract:

Objective      To determine the effect of long non-coding RNA (lncRNA) Gm4419 on the proliferation and fibrosis of mouse glomerulus mesangial cells under high glucose condition. Methods       The expression of Gm4419 in the kidney tissues from the mouse model of diabetic nephropathy (DN) and glomerulus mesangial cells cultured under high glucose condition were detected by fluorescence qRT-PCR. Gm4419 siRNAs were designed and synthesized, and overexpression plasmid of pcDNA3.1 (+)-Gm4419 was constructed. Then the Gm4419 siRNAs and pcDNA3.1 (+)-Gm4419 over-expression plasmids were transiently transfected respectively into the glomerulus mesangial cells under high or low glucose condition. EdU assay was used to detect the proliferation ability of the transfected cells. Western blotting was employed to detect the protein levels of renal fibrosis markers. Results       The expression levels of Gm4419 were significantly higher in the kidney tissues of DN mice and high-glucose cultured glomerulus mesangial cells when compared with the tissues of control mice and low-glucose cultured cells (P<0.01). After the Gm4419 siRNA with best interfering effect screened by fluorescence qRT-PCR was transfected into the high-glucose cells, the knockdown resulted in the decrease in the proliferation and fibrosis in the cells when compared with high glucose mock cells or control cells (P<0.05). In addition, after the over-expression plasmid pcDNA3.1 (+)-Gm4419 confirmed by enzyme digestion and DNA sequencing was transfected into the cells cultured under low glucose condition, the proliferation ability and fibrosis were enhanced compared with low glucose mock cells or control group (P<0.05). Conclusion       lncRNAGm4419 is involved in the regulation of the proliferation and fibrosis of mesangial cells from DN mice.

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更新日期/Last Update: 2017-02-05