Influence Of Leflunomide On Nuclear Factor-κB And Tumor Necrosis Factor-α In Diabetic Nephropathy Rats

Objective To investigate the expression of nuclear factor-κB(NF-κB) and tumor necrosisfactor-α(TNF-α),the prevention and cure effects of leflunomide in diabetic nephropathy rats.Method Thirty fale Wistar rats were randomly divided into three groups:control group;diabetic nephropathy(DN) group;diabetes treated with leflunomide group.After removing the left renal of rats,rat models of DN were created by injecting STZ(35mg/kg),while the controlling rats received identical volum of citrate buffer solution.When the model were established,rats in the leflunomide-treated were daily garvage of leflunomide(5mg/kg).Give the same capacity saline to comparison group and DN group.When the model were established, blood glucose and body weight were measured every two weeks.All 24 hours urine volume and 24 hours urine protein were determined at the end of eight and twelve weeks.Blood examples were collected from heart to detect serum creatinine and urea nitrogen.The pathological changes of kidney were observed in electron microscope and Hematoxylin and Eosin(HE) stain.The expression of NF-κB protein was examined by immunohistochemistry.The expression of TNF-αwas determined by enzyme linked immunosorbent assay(ELISA).Results(1) Model of DN is only a minor mode,and its rate of madding model was 95%. Itcango on being experimented by the availability of DN model added to experiment.After the major model,there is no death and its mortality is 0%.(2) With the extending of the course,the in weight of kidney of the rats of DN group increased,and creatinine clearance rate increased,and there is proteinuria.Compared with the control group,there were significant differences(P<0.05).The urine protein,serum creatinine,urea nitrogen of the rats of Leflunomide group decreased compared with the same period DN group significantly,and there were significant differences(P<0.05).(3) From histopathology observation the thickening of glomerular basementmembrane (GBM) of leflunomide group reduced significantly,compared with DN group,and mesangial cell proliferation and extracellular matrix(ECM) deposition reduced,which explained that leflunomide can ameliorate renal injury.(4) IHC showed that the expression of NF-κB increased significantly in DNgroup.Compared with the DN group,NF-κB attenuated in DN treated with leflunomide(P<0.05).With the extending of the course,leflunomide treatment inhibited the protein levels of NF-κB in the glomerulus and tubulonterstitium of DN model as dementrated by IHC(P=0.013).(5) ELISA analyses showed the expression of TNF-αincreased significantly in DNgroup, compared with control group.Compared with the DN group,TNF-αreduced in DN treated with leflunomide(P<0.05).With the extending of the course,leflunomide treatment inhibited the protein levels of TNF-αin serum of rats of DN model.Conclusion NF-κB,TNF-αmay play a negative role in the progression of DN.Leflunomide may protect kidney injury through inhibiting the protein levels of NF-κB and TNF-α.