發(fā)布時(shí)間：2018-03-04 01:16

  本文選題：長(zhǎng)鏈非編碼RNA　切入點(diǎn)：漿細(xì)胞瘤異位基因　出處：《山東醫(yī)藥》2017年14期 　論文類型：期刊論文

【摘要】：目的觀察長(zhǎng)鏈非編碼RNA(lncRNA)中的漿細(xì)胞瘤異位基因1(PVT1)表達(dá)下調(diào)對(duì)高糖環(huán)境下腎小球系膜細(xì)胞增殖及細(xì)胞外基質(zhì)(ECM)相關(guān)蛋白表達(dá)的影響,探討PVT1在糖尿病腎病發(fā)病中的作用機(jī)制。方法培養(yǎng)人腎小球系膜細(xì)胞,分為正常對(duì)照組、高糖組、高糖+control siRNA組、高糖+PVT1 siRNA組。正常對(duì)照組在5.55 mmol/L的低糖DMEM完全培養(yǎng)基中培養(yǎng),高糖組、高糖+control siRNA組、高糖+PVT1 siRNA組均在25mmol/L的高糖DMEM完全培養(yǎng)基中培養(yǎng)。高糖+control siRNA組、高糖+PVT1 siRNA組分別轉(zhuǎn)染control siRNA、PVT1 siRNA。分別培養(yǎng)0、24、48、72 h后,采用CCK-8法測(cè)算各組細(xì)胞增殖率。采用流式細(xì)胞術(shù)檢測(cè)各組細(xì)胞G1、S期比例。采用qRT-PCR法檢測(cè)各組細(xì)胞PVT1 mRNA表達(dá)。采用Western blot法檢測(cè)各組細(xì)胞ECM相關(guān)蛋白纖維連接蛋白(FN)、轉(zhuǎn)化生長(zhǎng)因子β_1(TGF-β_1)及Ⅰ型纖溶酶原激活物抑制因子(PAT-1)蛋白的表達(dá)。結(jié)果高糖組細(xì)胞各時(shí)間點(diǎn)細(xì)胞增殖率均高于正常對(duì)照組,高糖+PVT1 siRNA組各時(shí)間點(diǎn)細(xì)胞增殖率均低于高糖組(P均0.05)。與正常對(duì)照組比較,高糖組細(xì)胞G1期比例減少、S期比例增多;與高糖組比較,高糖+PVT1 siRNA組G1比例增加、S期比例減少(P均0.05)。高糖組、高糖+control siRNA組PVT1 mRNA相對(duì)表達(dá)量均高于正常對(duì)照組,高糖+PVT1 siRNA組PVT1 mRNA相對(duì)表達(dá)量低于高糖組(P均0.05)。高糖組、高糖+control siRNA組、高糖+PVT1 siRNA組FN、TGF-β_1、PAT-1蛋白表達(dá)均高于正常對(duì)照組,高糖+PVT1 siRNA組FN、TGF-β_1、PAT-1蛋白表達(dá)均低于高糖組(P均0.05)。結(jié)論在高糖環(huán)境下,腎小球系膜細(xì)胞中PVT1表達(dá)明顯增加,下調(diào)PVT1表達(dá)可明顯抑制腎小球系膜細(xì)胞的增殖、抑制ECM相關(guān)蛋白的表達(dá)。
[Abstract]:Objective to investigate the effect of down-regulation of plasmacytoma ectopic gene 1pPVT1 on the proliferation of glomerular Mesangial cells and the expression of ECM related proteins in high glucose environment. Methods Human glomerular Mesangial cells were cultured and divided into normal control group, high glucose group and high glucose control siRNA group. High glucose PVT1 siRNA group was cultured in 5. 55 mmol/L low glucose DMEM complete medium, high glucose group, high glucose control siRNA group, high glucose PVT1 siRNA group were all cultured in 25 mmol / L high glucose DMEM complete medium, high glucose control siRNA group, high glucose control siRNA group, high glucose control siRNA group, high glucose control siRNA group, high glucose control siRNA group, high glucose PVT1 siRNA group were all cultured in 25 mmol / L high glucose DMEM complete medium. High glucose PVT1 siRNA group was transfected with control siRNA-PVT1 siRNA. The proliferation rate of each group was measured by CCK-8 method, the proportion of G1S phase was detected by flow cytometry, the expression of PVT1 mRNA was detected by qRT-PCR assay, and the ECM associated protein fibronectin fibronectin was detected by Western blot assay. Expression of TGF- 尾 1) and plasminogen activator inhibitor type I (PAT-1) protein in TGF- 尾 1. Results the cell proliferation rate in high glucose group was higher than that in normal control group at all time points. The cell proliferation rate of high glucose PVT1 siRNA group was lower than that of high glucose group at each time point (P < 0.05). Compared with the normal control group, the G 1 phase ratio of high glucose group was lower than that of high glucose group, and the proportion of S phase was increased in high glucose group. The relative expression of PVT1 mRNA in the high glucose group and high glucose control siRNA group was higher than that in the normal control group, and the relative expression of PVT1 mRNA in the high glucose PVT1 siRNA group was lower than that in the high glucose PVT1 siRNA group, and the relative expression level of PVT1 mRNA in the high glucose PVT1 siRNA group was lower than that in the high glucose group. In high glucose control siRNA group and high glucose PVT1 siRNA group, the expression of FN- TGF- 尾 1 PAT-1 protein was higher than that in normal control group, and the expression of FN- TGF- 尾 1 PAT-1 protein in high glucose PVT1 siRNA group was lower than that in high glucose group (P 0.05). Conclusion the expression of PVT1 in glomerular Mesangial cells increased significantly in high glucose environment. Down-regulation of PVT1 expression significantly inhibited the proliferation of glomerular Mesangial cells and the expression of ECM related proteins.
【作者單位】： 廣州醫(yī)科大學(xué)附屬第二醫(yī)院;
【基金】：廣東省省級(jí)科技計(jì)劃資助項(xiàng)目(2014A020212324)
【分類號(hào)】：R692

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