Effect Of Yishen Huoxue Qingxie Recipe On Renal Fibrosis Based On TGF-β1/Smad3 Pathway

Objective: To observe the clinical efficacy and safety of Yishen Huoxue Qingxie Decoction in the treatment of chronic kidney disease(CKD)stage 3-5 non dialysis patients;to explore the effect of Yishen Huoxue Qingxie Decoction on renal fibrosis and TGF-β1 /Smad3 signaling pathway;to explore effective prescriptions,and to provide ideas for the research of related traditional Chinese medicine in the prevention and treatment of renal fibrosis.Methods:1.Clinical trial:According to the requirements of clinical trial design,72 patients were selected and randomly divided into treatment group and control group,36 cases in each group.The treatment group was treated with Yishen Huoxue Qingxie Decoction for 8weeks.The changes of laboratory indexes(serum creatinine,urea nitrogen,uric acid,hemoglobin,urinary protein / creatinine ratio,glomerular filtration rate,serum TGF-β1,Smad3 levels),TCM symptoms and syndrome scores were observed before and after treatment.2.Cell experiment:Normal human renal tubular epithelial cell line(HK-2)was cultured in vitro,and HK-2 epithelial mesenchymal transition(EMT)model was induced by cyclosporine A(CsA).The experiment was divided into normal control group(group A),model group(group B),valsartan control group(Group C),low-dose Yishen Huoxue Qingxie recipe group(Group D),medium dose Yishen Huoxue Qingxie recipe group(Group E)and high-dose Yishen Huoxue Qingxie recipe group(Group F).MTT assay was used to detect the cell viability,flow cytometry and scratch test were used to detect the cell apoptosis rate and migration ability,and Western blot was used to detect the protein expression levels of TGF-β1 and Smad3.Results:1.Clinical trial results:After treatment,the TCM syndrome scores of the two groups were decreased,especially in the treatment group(P < 0.05),and the total effective rate of TCM syndromes in the treatment group and the control group were 69.70% and43.75%respectively,the treatment group was better than the control group(P < 0.05);After treatment,the total effective rate of the treatment group and the control group were 78.78%and 59.37%,respectively,and the treatment group was better than the control group(P <0.05).After treatment,therapeutic efficacy related indexes SCR,BUN,UA,HGB,UACR,EGFR,TGF-β1 and Smad3 were improved in the treatment group,SCR and EGFR were not significantly improved in the control group,but BUN,UA,HGB,UACR,TGF-β1 and Smad3 were improved.The treatment group was more effective in improving BUN,UA,UACR,TGF-β1 and Smad3 than the control group(P < 0.05).2.Cell experiment results:(1)cell viability and apoptosis rate:After CsA treatment,cell viability decreased significantly and cell apoptosis rate increased.After intervention with valsartan and high,medium and low doses of Yishen Huoxue Qingxie decoction,cell viability in each group increased in varying degrees(P < 0.05),and cell apoptosis rate decreased(P < 0.05);the improvement of C,D and E in group C was better than that in group F(P < 0.05),and there was no statistical difference between the groups(P > 0.05).(2)Effect of migration ability: compared with group A,the migration ability of group B,C,D,E and F was enhanced(P < 0.05),in which group B was significantly enhanced(P < 0.05);the migration inhibition ability of group E was higher than that of group C and F(P < 0.05),and there was no significant difference between group E and group D(P > 0.05).(3)The protein expression of TGF-β1: group B,C,D,E and F was up-regulated in varying degrees(P < 0.05),in which group B was up-regulated significantly(P < 0.05);the protein expression of group D was lower than that of group C,e and f(P < 0.05).(4)Smad3:compared with group A,the protein expression of group B was significantly up-regulated(P < 0.05);compared with group B,the protein expression of group C,D,E and F was decreased(P < 0.05),and the protein expression of group E was lower than that of group C,D and F(P < 0.05).Conclusion:1.Yishen Huoxue Qingxie decoction can improve the clinical symptoms and renal function of CKD3-5 patients with spleen kidney deficiency and Turbid Toxin stasis syndrome,and reduce urinary protein excretion.2.Yishen Huoxue Qingxie decoction can reduce the expression of TGF-β1 and Smad3 in serum of CKD3-5 patients,and delay renal fibrosis.3.Yishen Huoxue Qingxie recipe can effectively inhibit the transdifferentiation of human renal tubular epithelial cells induced by cyclosporine A by partially blocking the TGF-β1 / Smad3 signaling pathway,and play an anti renal fibrosis role.