基础医学与临床 ›› 2016, Vol. 36 ›› Issue (11): 1505-1510.

• 研究论文 • 上一篇    下一篇

调控Nampt对DN肾小球细胞表达vimentin的影响

陈叶1,王曼伊2,胡婷婷1,黄爽1,冯乔2,王平1,冯乐平1   

  1. 1. 桂林医学院
    2. 桂林医学院附属医院
  • 收稿日期:2016-05-26 修回日期:2016-09-20 出版日期:2016-11-05 发布日期:2016-10-24
  • 通讯作者: 冯乐平 E-mail:fqw12350@sina.com
  • 基金资助:
    国家自然科学基金;广西科技厅自然科学基金

Effect of Vimentin expression by Regulated Nampt in glomerular cells in diabetic nephropathy

  • Received:2016-05-26 Revised:2016-09-20 Online:2016-11-05 Published:2016-10-24
  • Contact: leping feng E-mail:fqw12350@sina.com

摘要: 目的 探讨肾小球细胞内源性Nampt对vimentin表达的影响和作用机制。方法 用C57/BL6 糖尿病肾病(DN)小鼠肾脏组织经包埋、固定、切片,经免疫共聚焦技术分析内源性Nampt和vimentin蛋白的表达和定位关系;在200 mmol/L葡萄糖(高糖)培养肾小球膜HBZY -1细胞至第5 d时,分别用10 μmol/L FK866和1 mmol/L NMN干预,通过免疫荧光和免疫印迹方法检测内源性Nampt对NF-κB、SIRT1和vimentin表达;按高糖不同培养时间(24、48、72、96、120和144 h)分组,检测细胞Nampt、NF-κB、SIRT1表达时间效应关系。结果 1) DN组小鼠肾小球明显萎缩,肾小球细胞内Nampt、vimentin 蛋白表达明显增加(P﹤0.01);2) 应用FK866处理细胞,vimentin表达明显减少(P﹤0.01);用NMN干预后,vimentin表达同样低于相应对照组(P﹤0.01);3) 随高糖培养细胞时间延长,Nampt和NF-κB表达明显增加(P﹤0.01),相反SIRT1 表达减少(P﹤0.01)。结论 在DN肾小球纤维化中,Nampt能够通过增强NF-κB和抑制SIRT1信号途径,影响肾小球内vimentin蛋白表达。

关键词: 关键词:糖尿病肾病, Nampt, NF-κB, Sirt1, Vimentin

Abstract: Objective To study the function of endogenous Nampt on vimentin expression in glomerular cells. Methods With C57/BL6 diabetic mice experiment, the renal pathological tissue bebing embedded, fixated and sliced, endogenous Nampt and vimentin expression and location are analysized by appling confocal microscope technique. After 5 days in cultured with 200 mmol/L high concentration glucose (hyperglucose), HBZY-1 cells intervened with 10 μmol/L FK866 and 1 mmol/L NMN, Nampt, NF-κB, Sirt1 and vimentin are measured by immunofluorescence and Western blot assay respectively. HBZY-1 cells were divided into groups (24 h,48 h,72 h,96 h,120 h and 144 h) after cultured with hyperglucose, The time-effect course on expression of endogenous Nampt, NF-κB and Sirt1are measured by Western blot assay. Results 1) With glomerular atrophy, excess vimentin expression clearly increased along with Nampt high expression in DN glomerular cells, as compared with the control group (P﹤0.01). 2) After the cells treated with FK866, vimentin expression decreased significantly following Nampt expression inhibited (P﹤0.01). With NMN intervention, vimentin expression is very lower than that of control group (P﹤0.01). 3) With the extension of incubation time of the in the HBZY-1 cells in high concentrations glucose, the expression of Nampt and NF-κB obviously increased (P﹤0.01), while the SIRT1 expression decreased significantly, as compared with control groups (p﹤0.01). Conclusion In DN glomerular fibrosis, Nampt can control vimentin expression by enhenced NF-κB and inhibited SIRT1signaling pathway.

Key words: Key words: Diabetic Nephropathy, Nampt, NF-κB, Sirt1, Vimentin

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