lncRNA-Gm4419对高糖培养的肾小球系膜细胞增殖和纤维化的影响

发布时间：2018-11-22 08:50

【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,Ed U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(P0.01)。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(P0.05)。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(P0.05)。结论lnc RNA-Gm4419参与糖尿病肾小球系膜增生和纤维化的调节。
[Abstract]:Objective to investigate the effects of long chain non-coding Gm4419 on proliferation and fibrosis of glomerular Mesangial cells in mice cultured with high glucose. Methods fluorescence quantitative PCR was used to detect the expression of Gm4419 in diabetic kidney and glomerular Mesangial cells cultured with high glucose. PcDNA3.1 ()-Gm4419 overexpression plasmids were constructed by designing and synthesizing Gm4419 siRNA, and Gm4419 siRNA and pcDNA3.1 ()-Gm4419 overexpression plasmids were transfected into glomerular Mesangial cells cultured with high and low glucose, respectively. The proliferation ability of transfected cells was detected by Ed U method. The expression of fibronectin in kidney was detected by Western blot. Results the expression of Gm4419 in diabetic kidney and glomerular Mesangial cells cultured with high glucose was significantly higher than that in normal and low glucose groups (P0.01). Compared with high glucose mock or control group, compared with high glucose mock group or control group, the proliferation ability of high glucose Gm4419 knockdown group was decreased and the expression of fibrosis marker protein was decreased (P0.05). In addition, the results of enzyme digestion and sequencing confirmed that the constructed pcDNA3.1 ()-Gm4419 overexpression plasmid was transfected into the cells of low glucose group. Compared with the low glucose mock group or the control group, the proliferation ability of the cells in the low glucose Gm4419 overexpression group was enhanced. The expression level of fibrosis marker protein was increased (P0.05). Conclusion lnc RNA-Gm4419 is involved in the regulation of glomerular Mesangial hyperplasia and fibrosis in diabetic patients.
【作者单位】：重庆医科大学基础医学院细胞生物学及遗传学教研室;重庆医科大学基础医学院生物信息学教研室;第三军医大学高原军事医学系军事医学地理学教研室;重庆医科大学基础医学院实验教学中心;
【基金】：国家自然科学基金预研项目(NSFYY201524) 重庆医科大学基础医学院2015年苗圃基金(JC201508)~~
【分类号】：R692.9;R587.2

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