| Objective: To study the effect of endogenous Nampt on Vimentin expression by regulated glomerular fibrosis in diabetic mice model via regulating NF-kappa B and Sirt1. Methods: Applicating diabetic animal models and preparation of kidney biopsy, expression and position of endogenous Nampt and VIM were evaluated in renal glomerular cell both by immune confocal and immune fluorescent microscope. Then, using HBZY-1 cell line, the proteins expression were measured respectively with high concentration of glucose(200mmol/L) of oxidative stress. The experimental group as follows:(1) group(200 mmol /L high glucose),(2) group(200mmol/L glucose + FK866),(3)group(200mmol/L glucose + NMN),(4) group(200mmol/L glucose + FK866 +NMN);The control group:(5)group(0.56mmol/L glucose),(6)group(0.56mmol/L glucose + FK866)(7) group(0.56mmol/L glucose + NMN). In vitro test, all Nampt, NF-κB, Sirt1 and VIM expression were detected by immunofluore-scence assay and those proteins were further confirmed again with western blot techniques. Results: All glomerulars showed obvious atrophy in diabetic groups compared with control group’s. VIM expression obvious increased in the glomerular cells while Nampt expression was significantly increased in the same cells. The protein expression of Nampt and VIM were obvious increased in glomerular mesangial cells HBZY-1 cell after oxidative stress in high glucose treatment(0-5d), at the same times, NF-kappa B protein expression increased but Sirt1 expression significantly reduced in cells(p﹤0.01). When endogenousNampt expression was inhibited by FK866, VIM expression was also obvious decreased following the changes of NF-kappa B and Sirt1 expression. The same results were also seen with NMN treatment. Conclusion: Endogenous Nampt may obviously through the NF-kappa B and Sirt1 interaction to affect VIM expression, which could affect the process of inflammation of kidney fibrosis in diabetic nephropathy. |
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