Nampt Regulate NF-κBp65 And Sirt1 And The Interaction Relationship In The Pathogenesis Process Of Diabetic Nephropathy

Objective: To investigate Nampt, NF-κBp65 and Sirt1 expression and the interaction relationship between them by Western blot, immunofluorescence and immunohistochemistry techniques in the diabetic nephropathy tissues(eg. pancrease and kidney). It is pave the way for the diabetic nephropathy treatment. Methods: 1. Tubular epithelial NRK-52 E cells routinely cultured in RPMI-1640 medium under oxidation stress with high glucose concentration(35mmol/L) for different time(0h, 24 h, 48 h, 72 h, 96 h, 120h). The Nampt, NF-κBp65 and Sirt1 protein expression are detected by western blot. In the training process, cell morphology was observed, meanwhile, the expression of Nampt,NF-κBp65 and Sirt1 gene expression are reconfirmed by immunofluorescence. 2. Nampt,NF-κBp65 and Sirt1 expression in pancreas and kidney cell are also detected in vivo test with type 2 diabetes model of SD rat by immunohistochemistry assay. Results: 1.high concentration of glucose(35mmol/L) 1640 RPMI-1640 medium NRK-52 E cells, the application to detect the expression levels of Nampt and NF-κBp65 protein increased while Sirt1 protein level reduced with the increasing of culture time by western blot;Compared with the normal glucose concentration(11mmol/L) group, high glucose concentration(35mmol/L) grop cultured cells becomes larger, irregular and significant morphological changes.By immunofluorescence, Nampt expression was significantly increased and was expressed in the cytoplasm and nucleus,while expression of NF-κBp65 also significantly increased but Sirt1 decreased.2. In the pancreas of SD rat model of type 2 diabetic,Nampt, NF-κBp65 and Sirt1 protein mainly in islets site, Nampt and NF-κBp65 was significantly higher than the control group, Sirt1 protein was lower than the control group; in the kidney tissue, Nampt, NF-κBp65 and Sirt1 protein mainly expressed in renal tubular and part of the glomerular,Nampt and NF-κBp65 was significantly higher, while Sirt1 protein expression was lower than the control group. Conclusions: 1. Cultured the NRK-52 E cells with high glucose concentrations,the expression of protein Nampt and NF-κBp65 increased and Sirt1 reduced. 2. In the pancreas and kidney of the SD rat of type 2 diabetes model, the expression of Nampt and NF-κBp65 increased and the expression of Sirt1 decreased compared with the normal group.In the pathogenesis of diabetes and diabetic nephropathy, Nampt may play an important role in the pathogenesis of diabetes and diabetic nephropathy by regulating the expression of NF-κBp65 and Sirt1.