调控Nampt对糖尿病肾病肾小球细胞表达波形蛋白的影响  

Effect of regulating Nampt on vimentin expression in glomerular cells in diabetic nephropathy

作　　者：陈叶[1] 王曼伊 胡婷婷[3] 黄爽[1] 冯乔[2] 王平[1] 冯乐平[1] 

机构地区：[1]桂林医学院公共卫生学院,广西桂林541004 [2]桂林医学院附属医院优生遗传科,广西桂林541004 [3]桂林医学院生物技术学院,广西桂林541004

出　　处：《基础医学与临床》2016年第11期1505-1510,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(81460164);广西科技厅自然科学基金(2015GXNSF)

摘　　要：目的探讨肾小球细胞内源性Nampt对波形蛋白(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10μmol/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和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。Objective To study the function of endogenous Nampt on vimentin expression in glomerular cells.Methods C57 / BL6 diabetic mice renal pathological tissue was embedded,fixated and sliced,endogenous Nampt and vimentin expression and location are analysized by appling confocal microscope technique. After 5 days of incubation 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（ 24,48,72,96,120 and 144 h） after cultured with hyperglucose. The time-effect course on expression of endogenous Nampt,NF-κB and Sirt1 are measured by Western blot assay. Results 1） With glomerular atrophy,excess vimentin expression clearly increased along withNampt high expression in DN glomerular cells,as compared with the control group（ P〈0. 01）. 2） After the treatment with FK866,vimentin expression significantly decreased 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）. Conclusions In DN glomerular fibrosis,Nampt can control vimentin expression by enhenceing NF-κB and inhibiting Sirt1 signaling pathway.

关 键 词：糖尿病肾病 NAMPT NF-κB 沉默子Sir2同源体1 波形蛋白 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]