Study On The Activation Of HGF/c-met Pathway And The Effect Of Different Ingredients Of Liuwei Dihuang Decoction In5/6Nephrectomy Rats

Objective:1. To study the activation of HGF/c-met pathway in5/6neph-rectomy rats and the treatment mechanism of Liuwei Dihuang Decoction.2.To study the protectionof different combinations of Liuwei Dihuang Decoction on kidney function in5/6Nephrectomy rats.Methods:Experiment1:The60healthy male Sprapue-Demey rats were randomly divided into normal group, sham operation group, model group and Liuwei Dihuang decoction group. Model group and Liuwei Dihuang decoction group both adopted5/6nephrectomy. A week after operation. Liuwei Dihuang decoction group were administrated with Liuwei Dihua-ng decoction, other groups all received equivoluminal distilled water. All animals were killed after8weeks. The24hour urinary protein, serum creatinine and blood ureanitrogen in rats were tested, the remnant kidney were weighted, the pathological changes were observed, the expression level of protein and mRNA of HGF、c-met and Col-Ⅳ in reremnant kidney were measured.Experiment2:The60healthy male Sprapue-Demey rats were randomly divided into Liuwei Dihuang decoction group, three tonifying ingredients group and three reducing ingredients group.Used the same way to set up rat model. A week after operation,Liuwei Dihuang decocti-on group were administrated with Liuwei Dihuang decoction, three tonifying ingredients group were administrated with three tonifying ingredients of Liuwei Dihuang decoction(Ehmannia,Fructus corni, Chine-se yam),and three reducing ingredients were administrated with three reducing ingredients group of Liuwei Dihuang decoction(Rhizoma alismatis, Poria cocos, Cortex Moutan). All animals were killed after8weeks. The observation methods are the same as experiment1.Results:l.After5/6neSphrectomy, the remnant kidney tissues of rats appeared pathological changes,such as glomerular sclerosis, renal tubular lesions and interstitial fibrosis, the levels of serum creatinine, blood urea-nitrogen and24h urinary protein all raised(P<0.01).After the intervene-tion of Liuwei Dihuang decoction, the pathological changes of remnant renal tissues improved, the levels of serum creatinine, blood ureanitrogen and24h urinary protein all obviously reduced (P<0.01)2. After5/6nephrectomy, the expression level of protein of HGF、 c-met and Col-Ⅳall increased in the remnant renal tissues of rats (P<0.01). After the intervention of Liuwei Dihuang decoction,the expression level of HGF protein and c-met protein levels further increased(P<0.05), while the expression level of Col-Ⅳprotein decreased (P<0.05). After5/6nephrectomy, the expression level of of HGFmRNA increased in the remnant renal tissues of rats (P<0.05), the expression level of c-met mRNA and Col-Ⅳ mRNA also further increased (P<0.01). After the intervention of Liuwei Dihuang decoction, the expression level of HGFmRNA further increased (P<0.01), the expression level of c-metmRNA further increased too (P<0.05), however,the expression level of Col-IV mRNA decreased (P<0.05)3. After the intervention of three tonifying ingredients of Liuwei Dihuang decoction,and three reducing ingredients of Liuwei Dihuang dec-oction, the pathological changes were more serious in the remnant renal tissues of rats, the levels of serum creatinine,blood urea nitrogen and24h urinary protein are higher than Liuwei Dihuang decoction group, the expression level of HGF protein, HGF mRNA, c-met protein and c-met mRNA are all less than Liuwei Dihuang decoction group,however, the expression level of Col-Ⅳ protein and Col-Ⅳ mRNA are higher than Liuwei Dihuang decoction group.Conclusion:1. After5/6nephrectomy, the remnant renal tissues of rat appeared pathological changes,such as glomerular sclerosis, renal tubular lesions and interstitial fibrosis, the levels of serum creatinine、 blood urea nitrogen and24h urinary protein all raised. Suggested success-fully copied5/6nephrectomy rat model.2. Liuwei Dihuang decoction can reduce pathological changes and improve renal function in5/6nephrectomy rats. 3. After5/6nephrectomy, HGF/c-met pathway was activated in the remnant renal tissues of rats.4. Liuwei Dihuang decoction can reduce the expression level of Col-Ⅳ, alleviate renal fibrosis. Which might be relative to the function that Liuwei Dihuang decoction can improve the expression level of HGF/c-met in the remnant renal tissues of rats.5. Liuwei Dihuang decoction can reduce pathological changes and protect renal function in5/6nephrectomy rats. Its roles were better than th-Ree the tonifying ingredients of Liuwei Dihuang decoction and the three reducing ingredients of Liuwei Dihuang decoction.