Study On The Mechanism Of Fushengong Decoction Against Renal Interstitial Fibrosis In Rats With Chronic Renal Failure

BackgroundChronic Renal Failure?CRF?is the final destination of the continuous progress of various chronic kidney diseases?CKD?.As one of the global public health problems,the whole world's medical experts of kidney disease have been thinking and making a lot of experimental research to solve this problem thoroughly.At present,about 2.4 million patients with renal failure in the world need dialysis.The annual number of patients with CRF increases by 7-8%.End-stage renal disease can only be maintained by dialysis and kidney transplantation.The cost is quite costly and leads to many end-stage patients can't afford the financial burden and abandon treatment.Traditional Chinese medicine for the prevention and treatment of CRF has the advantages of accurate curative effect,safe medication and low cost.It has been paid more and more attention,and Fushengong decoction?FSGD?with effectivity belongs to Guoziguang professor who is famous specialist of traditional chinese medicine and the national medical master was used more than 60 years in the treatment of CRF.The clinical effect of stage?and?was affirmed by the majority of patients.Therefore,this study started with the expression of klotho protein,mRNA and Wnt/?-catenin signal pathway as point of penetration and studied the mechanism of FSGD against renal interstitial fibrosis in rats with CRF,in order to clarify its possible mechanism of delayed progression of CRF.ObjectiveIn view of the research results of the previous research group,to further explore the possible mechanism of FSGD in treating CRF,this experiment intends to further explore the effects of this formula on CRF rats from the perspective of reduction and repair of renal injury and against renal interstitial fibrosis.The expression of effect of klotho protein and mRNA and Wnt/?-catenin signal pathway provide a reliable laboratorial basis for further excavating the clinical experience of Chinese medicine masters and developing a Chinese patent medicine with a clear mechanism,safety,effectiveness,convenience and affordability.MethodsFifty-five Sprague-Dawley male rats were selected and after one week of adaptive feeding,they were randomly divided into the normal group,the model group and the low,medium and high-dose group of FSGD,with 11rats in each group.The normal group was given conventional breeding,and the remaining 4 groups of rats were fed a 0.5%adenine feed to establish a CRF model and fed for 3 weeks.After the modeling was completed,4 rats of model groups and 1 normal rat were randomly selected for hematuria biochemical indicators and histopathological examination to prove whether the modeling is success.After successful modeling,all rats were fed with conventional feed.Rats in the normal group and model group were administered with 0.9%normal saline?NS?20 mL_·kg_·^-1d^-1;rats in the low,medium and high-dose groups of FSGD were dosed according to body weight 4,8,16 g_·kg^-11 FSGD decoction was administered to the stomach,once a day for 30 days.After the intervention treatment,20%urethane 5mL_·kg-1 body weight was used for abdominal anesthesia and 3-5 mL of blood was collected from the heart.The serum was separated to collect supernatant and stored at 4?for future use.Then the rats were sacrificed by cervical dislocation.The abdominal cavity was opened to expose the bilateral kidneys.Hemostatic forcep was used to fix in and out of the renal arteries and veins to prevent a large amount of blood from flowing into the abdominal cavity.They were fixed with 4%paraformaldehyde and frozen with liquid nitrogen.The kidneys after paraformaldehyde fixation were mainly used for paraffin embedding.The kidneys after liquid nitrogen quick-frozen were transferred to-80?refrigerator and stored for future use.Serum of rats in each group was taken to detect the levels of serum creatinine?SCr?and urea nitrogen?BUN?.HE and Masson staining were used to detect the pathological and morphological changes of rat kidney tissues.The mRNA expression of Klotho,Wnt4,and?-catenin were detected by RT-PCR.The protein expression of Klotho,Wnt4,?-catenin were detected by Western Blot and immunohistochemical in kidney tissues.ResultsCompared with the normal group,the levels of SCr and BUN in the serum of the model group increased significantly?P<0.01?,the pathological damage of kidney tissues increased obviously and the expression of Klotho protein and mRNA in the kidney reduced significantly?P<0.01?.The expression of Wnt4,?-catenin protein and mRNA increased significantly?P<0.01?.Compared with the model group,the renal pathological damage of rats in each dose group of FSGD reduced significantly and the contents of serum SCr and BUN reduced significantly?P<0.05,P<0.01?.The expression of Klotho protein and mRNA in kidney tissues increased significantly?P<0.05?and the expressions of Wnt4,?-catenin protein and mRNA decreased significantly of rats in each dose group of FSGD?P<0.05?.ConclusionFSGD can reduce and repair renal damage effectively,inhibit renal interstitial fibrosis and delay the progression of CRF.The mechanism may be related to increasing the expression of renal Klotho protein and mRNA and reducing the expression of Wnt4,?-catenin protein and mRNA.