The Intervention Effect Of ShenKan Wan On Diabetic Nephropathy Fibrosis Based On EndMT

BackgroundDiabetic nephropathy(DN)is one of the most common and serious microvascular complications in patients with diabetes mellitus(DM).The pathogenesis of DN is very complex.In recent years,relevant studies have found that endothelial dysfunction can promote the development of DN.Fibroblasts produced during Endothelial-mesenchymal transition(EndMT)are one of the major sources of renal fibroblasts,whereas renal fibroblasts play a major role in renal fibrosis,While renal fibrosis is the main pathological manifestation of DN.Inflammatory factors,high glucose and so on will endothelium EndMT,which will gradually lead to all kidney cell fibrosis,the final loss of function of the kidney.Therefore,EndMT plays a crucial role in renal fibrosis,which is an important reason and outcome of endothelial cell injury-fibrosis.EndMT endothelial cells lose specific markers when they occur.Such as vascular endothelial cadherin(VE-cadherin),thereby regaining the specific markers of mesenchymal cells such as vimentin,a-smooth muscle actin,a-SMA)and so on.Therefore,EndMT may be an important mechanism leading to renal fibrosis,but its specific regulation is not yet clear.At the same time,endothelial cell injury fibrosis involves many molecular signaling pathways,of which Wnt/?-catenin can regulate the progress of fibrosis.Chinese medicine preparation Shenkangwan can improve DN to a certain extent and delay the progression of DN.However,there is no research yet that whether Shenkangwan can block EndMT and treat DN better is still unclear.Therefore,in this experiment,Shenkang pill was used to interfere with the expression of EndMT and Wnt/?-catenin signaling pathways in DN fibrotic mice and to explore the effect of Shenkang pill in the treatment of diabetic nephropathy fibrosis EndMT.ObjectiveTo observe the effect of Shenkangwan on EndMT and to explore the effect of Shenkangwan on EndMT in DN mice and its possible mechanism.Methods1.Animal groups:C57 mice aged 7-8 weeks were divided into normal control group,DN model group,high and low intervention group of traditional Chinese medicine,and irbesartan control group.Each group of ten mice,in addition to the normal control group of other mice DN modeling,modeling and success after Shen Kang pill and irbesartan gavage.2.testing the mice blood glucose,urea nitrogen,creatinine and other numerical conditions3.The HE and MASSON staining were used to observe the pathological changes of diabetic nephropathy and fibrosis,and the pathological changes of each group were observed.4.Immunohistochemistry and Western blot(WB)were used to detect the protein expression of a-sma,vimentin and VE-cadheri in the tissues.5.The mRNA and protein expression of Wntl,?-catenin and P-GSK-3?were detected by RT-PCR and WB,respectively.Results1.Compared with the normal control group,the blood glucose of DN model group increased obviously and the body weight decreased.The proteinuria of 24-hour urinary protein in the model group was significantly higher than that in the normal group(P<0.05).The proteinuria of 24-hour urinary protein in the Shenkang pill group was significantly lower than that in the model group(P<0.05)Compared with model group,BUN,Scr and Ccr significantly increased(P<0.05).The levels of BUN,Scr and Ccr in Shenkang pill group were significantly lower than those in model group(P<0.05).2.HE staining showed that the blank group showed normal renal tissue,the model group of kidney tissue cells hyperplasia,cell arrangement disorders,glomerular cells decreased.Irbesartan group,Shenkangwan low,medium and high doses of cells arranged in neat rows,no change in glomerular cells.Masson staining showed that the model group staining was strongly positive,bright green fibrous tissue flake proliferation,significant fibrosis;each treatment intervention group a small amount of fibrous tissue proliferation,degree of fibrosis decreased to varying degrees3.Immunohistochemistry and WB results suggest that the model group of a-sma,vimentin protein expression increased,VE-cadherin expression decreased.Compared with the model group,the expression of a-sma,vimentin and the expression of E-cadherin in irbesartan group and Shenkangwan low,medium and high dose groups decreased.4.The results of RT-PCR and WB showed that the mRNA and protein expressions of Wntl,P-GSK-3?,and ?-catenin in the model group were significantly increased compared with those in the normal group(p<0.05)The mRNA and protein expressions of Wntl,P-GSK-3p,and ?-catenin in the kidneys of Kangwan-treated mice were lower than those in the untreated group(P<0.05).There was no significant difference between irbesartan group and Shenkangwan group(P>0.05).ConslusionActivation of Wnt/?-catenin signaling pathway is involved in the pathogenesis of renal fibrosis in DN mice.Shenkang Pills can improve renal function and inhibit fibrosis in DN model mice.The mechanism may inhibit the expression of Wntl,?-catenin and P-GSK-3?,and inhibit the activation of Wnt/?-catenin pathway,thereby increasing the expression of VE-cadherin,decreasing the expression of a-SMA and Vimentin,To anti-fibrosis effect.Shenkangwan can inhibit the development of EndMT by regulating ?-catenin,which can promote the treatment of DN,prevent and delay the progression of renal fibrosis,and provide a new mechanism for the treatment of DN.