【摘要】：目的探討長鏈非編碼Gm4419對高糖培養(yǎng)的小鼠腎小球系膜細(xì)胞增殖和纖維化的影響。方法熒光定量PCR檢測Gm4419在糖尿病腎病腎臟組織及高糖培養(yǎng)的腎小球系膜細(xì)胞中的表達(dá)水平。設(shè)計(jì)合成Gm4419 siRNA,同時構(gòu)建pcDNA3.1(+)-Gm4419過表達(dá)質(zhì)粒,并將Gm4419 siRNA和pcDNA3.1(+)-Gm4419過表達(dá)質(zhì)粒分別轉(zhuǎn)染至高低糖培養(yǎng)的腎小球系膜細(xì)胞,Ed U法檢測轉(zhuǎn)染后細(xì)胞的增殖能力;Western blot檢測腎臟纖維標(biāo)記蛋白的表達(dá)水平情況。結(jié)果 Gm4419在糖尿病腎病腎臟組織及高糖培養(yǎng)的腎小球系膜細(xì)胞中的表達(dá)水平較正常組及低糖組顯著增高(P0.01)。熒光定量PCR篩選出一條最佳干擾Gm4419的siRNA,轉(zhuǎn)染高糖組后,與高糖mock組或?qū)φ战M比較,高糖Gm4419 knockdown組細(xì)胞增殖能力減弱、纖維化標(biāo)記蛋白表達(dá)減少(P0.05)。此外,將酶切和測序結(jié)果證實(shí)構(gòu)建成功的pcDNA3.1(+)-Gm4419過表達(dá)質(zhì)粒轉(zhuǎn)染至低糖組細(xì)胞,與低糖mock組或?qū)φ战M相比,低糖Gm4419過表達(dá)組細(xì)胞增殖能力增強(qiáng)、纖維化標(biāo)記蛋白表達(dá)水平增高(P0.05)。結(jié)論lnc RNA-Gm4419參與糖尿病腎小球系膜增生和纖維化的調(diào)節(jié)。
[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.
【作者單位】： 重慶醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院細(xì)胞生物學(xué)及遺傳學(xué)教研室;重慶醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院生物信息學(xué)教研室;第三軍醫(yī)大學(xué)高原軍事醫(yī)學(xué)系軍事醫(yī)學(xué)地理學(xué)教研室;重慶醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院實(shí)驗(yàn)教學(xué)中心;
【基金】：國家自然科學(xué)基金預(yù)研項(xiàng)目(NSFYY201524) 重慶醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院2015年苗圃基金(JC201508)~~
【分類號】：R692.9;R587.2

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