高浓度葡萄糖条件下肾小球系膜细胞内源性烟酰胺磷酸核糖转移酶对波形蛋白表达的调控作用

发布时间：2018-04-25 06:29

本文选题：高浓度葡萄糖 + 烟酰胺磷酸核糖转移酶　；参考：《吉林大学学报(医学版)》2017年05期

【摘要】：目的:探讨高浓度葡萄糖条件下肾小球系膜细胞过度表达的内源性烟酰胺磷酸核糖转移酶(Nampt)对波形蛋白(Vimentin)表达的调控作用,阐明糖尿病并发肾脏炎症纤维化的形成机制。方法:取患严重自发性糖尿病的C57/BL6小鼠肾脏,以野生型C57/L86小鼠作为对照组,行病理切片和组织荧光染色,应用免疫共聚焦方法检测肾小球系膜细胞中内源性Nampt和Vimentin表达及定位;肾小球膜HBZY-1细胞随机分为4组,即0.56mmol·L~(-1)低浓度葡萄糖(LG)对照组、200mmol·L~(-1)高浓度葡萄糖(HG)处理组、HG+FK866组和HG+NMN组;高浓度葡萄糖干预5d后,分别施加FK866(10μmol·L~(-1))和NMN(1 mmol·L~(-1))作用24h后,通过免疫荧光和免疫印迹法检测细胞内源性Nampt、Vimentin、核因子κB p65(NF-κBp65)和依赖于烟酰胺腺嘌呤二核苷酸(NAD+)的组蛋白去乙酰化酶1(Sirt1)表达水平;应用RT-PCR和免疫印迹等方法检测细胞Nampt和Vimentin表达水平。结果:与野生型C57/BL6小鼠组比较,严重糖尿病组小鼠肾小球明显萎缩,肾小球细胞内Vimentin表达水平随内源性Nampt表达水平升高而增加(P0.01);与0.56mmol·L~(-1)低浓度葡萄糖对照组比较,高浓度葡萄糖冲击细胞Nampt、NF-κBp65和Vimentin表达水平明显升高(P0.01),Sirt1表达水平明显降低(P0.01)。HG+FK866和HG+NMN组肾小球系膜细胞中Nampt、Vimentin和NF-κBp65表达水平均较对照组明显降低(P0.01)。结论:高浓度葡萄糖冲击条件下肾小球系膜细胞过度表达的内源性Nampt能够通过NF-κBp65和Sirt1信号通路促进Vimentin表达。
[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.
【作者单位】：桂林医学院公共卫生学院营养与食品卫生学教研室;桂林医学院附属医院优生遗传科;桂林医学院生物技术学院生物技术教研室;
【基金】：国家自然科学基金资助课题(81460164,31060161) 广西壮族自治区科技厅自然科学基金资助课题(2015GXNSF)
【分类号】：R587.2;R692.9

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