發(fā)布時(shí)間：2018-04-25 06:29

  本文選題：高濃度葡萄糖 + 煙酰胺磷酸核糖轉(zhuǎn)移酶��； 參考：《吉林大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)》2017年05期

【摘要】：目的:探討高濃度葡萄糖條件下腎小球系膜細(xì)胞過度表達(dá)的內(nèi)源性煙酰胺磷酸核糖轉(zhuǎn)移酶(Nampt)對波形蛋白(Vimentin)表達(dá)的調(diào)控作用,闡明糖尿病并發(fā)腎臟炎癥纖維化的形成機(jī)制。方法:取患嚴(yán)重自發(fā)性糖尿病的C57/BL6小鼠腎臟,以野生型C57/L86小鼠作為對照組,行病理切片和組織熒光染色,應(yīng)用免疫共聚焦方法檢測腎小球系膜細(xì)胞中內(nèi)源性Nampt和Vimentin表達(dá)及定位;腎小球膜HBZY-1細(xì)胞隨機(jī)分為4組,即0.56mmol·L~(-1)低濃度葡萄糖(LG)對照組、200mmol·L~(-1)高濃度葡萄糖(HG)處理組、HG+FK866組和HG+NMN組;高濃度葡萄糖干預(yù)5d后,分別施加FK866(10μmol·L~(-1))和NMN(1 mmol·L~(-1))作用24h后,通過免疫熒光和免疫印跡法檢測細(xì)胞內(nèi)源性Nampt、Vimentin、核因子κB p65(NF-κBp65)和依賴于煙酰胺腺嘌呤二核苷酸(NAD+)的組蛋白去乙�；�1(Sirt1)表達(dá)水平;應(yīng)用RT-PCR和免疫印跡等方法檢測細(xì)胞Nampt和Vimentin表達(dá)水平。結(jié)果:與野生型C57/BL6小鼠組比較,嚴(yán)重糖尿病組小鼠腎小球明顯萎縮,腎小球細(xì)胞內(nèi)Vimentin表達(dá)水平隨內(nèi)源性Nampt表達(dá)水平升高而增加(P0.01);與0.56mmol·L~(-1)低濃度葡萄糖對照組比較,高濃度葡萄糖沖擊細(xì)胞Nampt、NF-κBp65和Vimentin表達(dá)水平明顯升高(P0.01),Sirt1表達(dá)水平明顯降低(P0.01)。HG+FK866和HG+NMN組腎小球系膜細(xì)胞中Nampt、Vimentin和NF-κBp65表達(dá)水平均較對照組明顯降低(P0.01)。結(jié)論:高濃度葡萄糖沖擊條件下腎小球系膜細(xì)胞過度表達(dá)的內(nèi)源性Nampt能夠通過NF-κBp65和Sirt1信號通路促進(jìn)Vimentin表達(dá)。
[Abstract]:Aim: to investigate the effect of overexpression of endogenous nicotinamide phosphotransferase (Nampt2) on Vimentin expression in glomerular Mesangial cells under high glucose concentration and to elucidate the mechanism of renal fibrosis in diabetic patients. Methods: the kidneys of C57/BL6 mice with severe spontaneous diabetes were selected and wild-type C57/L86 mice were used as control group. The expression and localization of endogenous Nampt and Vimentin in glomerular Mesangial cells were detected by confocal immunoassay. Glomerular membrane HBZY-1 cells were randomly divided into 4 groups: 0.56mmol Lnln-1) low concentration glucose (LGG) control group (200mmol / L)) HG FK866 group and HG NMN group (treated with high concentration glucose for 5 days), and then treated with FK866(10 渭 mol L (1) and NMN(1 mmol L-1 (1) for 24 h, respectively. The expression of endogenous Namptt1 Vimentin, nuclear factor- 魏 B p65 (NF- 魏 Bp65) and histone deacetylase 1 (Sirt1) dependent on nicotinamide adenine dinucleotide (NAD) were detected by immunofluorescence and Western blotting. RT-PCR and Western blot were used to detect the expression of Nampt and Vimentin. Results: compared with the wild-type C57/BL6 mice group, the glomeruli of severe diabetic mice were significantly atrophied, the expression of Vimentin in glomerular cells increased with the increase of endogenous Nampt expression level, and compared with the control group of low concentration of 0.56mmol Lampan 1, the expression of Vimentin in glomerular cells increased with the increase of endogenous Nampt expression level. The expression level of NF- 魏 Bp65 and Vimentin in high glucose shock cells was significantly higher than that in the control group. The expression level of P0.01HG FK866 and HG NMN in glomerular Mesangial cells was significantly lower than that in the control group. The expression levels of Nampttl Vimentin and NF- 魏 Bp65 in glomerular Mesangial cells of HG NMN group were significantly lower than those of control group. Conclusion: overexpression of endogenous Nampt in glomerular Mesangial cells can promote Vimentin expression through NF- 魏 Bp65 and Sirt1 signaling pathway.
【作者單位】： 桂林醫(yī)學(xué)院公共衛(wèi)生學(xué)院營養(yǎng)與食品衛(wèi)生學(xué)教研室;桂林醫(yī)學(xué)院附屬醫(yī)院優(yōu)生遺傳科;桂林醫(yī)學(xué)院生物技術(shù)學(xué)院生物技術(shù)教研室;
【基金】：國家自然科學(xué)基金資助課題(81460164,31060161) 廣西壯族自治區(qū)科技廳自然科學(xué)基金資助課題(2015GXNSF)
【分類號】：R587.2;R692.9

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本文編號：1800139