Establishment And Preliminary Study Of The Mechanism Of The Combined Model Of Liver And Kidney Fibrosis

Since the declaration of Helsinki,cannot use human subjects to promote the progress of medical,therefore,some preclinical studies,such as Pharmacology,pharmacokinetics and mechanisms,can be based on the experimental animal ethics rules,using rodents and other animals with high similarity to human gene phenotypes to establish relevant models for research.Not only can controlled the variables and influencing factors artificially,but also can satisfy the research from the data quantity.It will be parallel to the human related diseases,so as to more convenient and effective research on the occurrence and development of human diseases,and further promote the progress of life science.Chronic liver disease and chronic kidney disease have been plaguing human beings for many years,especially in China.The number of patients with chronic kidney disease in China ranks first in the world.Meanwhile,chronic liver disease has been among the top five causes of death in China for a long time.Fibrosis is the necessary stage of all chronic diseases progress.This stage is widely considered to be reversible,and is regarded as a new target for curing chronic diseases of liver and kidney--to contain chronic diseases in the early stage of chronic diseases.However,at present,there is no effective treatment for liver and kidney fibrosis other than organ transplantation.Chronic liver disease and chronic kidney disease causes complex,single target drugs did not obtain good treatment effect,and the theory of traditional Chinese medicine "cure not ill",has its unique advantages in prevention and control of chronic diseases---system theoretical knowledge and rich clinical experience,multiple targets,so the theory of traditional Chinese medicine combined with traditional Chinese medicine is now seen as treatment of liver and kidney of chronic diseases,a new hope,But so far,there is no suitable animal model of liver and kidney fibrosis for scientific research to conduct extensive and in-depth research.This paper attempts to establish two kinds of liver and kidney fibrosis complex models,to carry out in-depth research and comparison on the characteristics and mechanism of the two kinds of liver and kidney fibrosis complex models.The first chapter reviews the methods used in the establishment of rodent models of liver and renal fibrosis by referring to domestic and foreign literatures.Through the different characteristics of different methods for the induction,comparative advantages and disadvant-ages of each model,provides a theoretical basis for the composite model of liver and kidney fibrosis,and also provides a scientific basis for the optimization of existing modeling methods and in-depth discussion of the pathogenesis of liver and kidney diseases.In the second chapter of this paper,TAA combined with UUO method was used to induce the composite model of liver and kidney fibrosis in SD rats.The experimental period was 4 weeks.The effects of model group and blank control group on the body weight and survival rate of rats were observed within 4 weeks.After 4 weeks,the rats were anesthetized and blood samples were collected to observe the serum indexes related to liver and kidney function:ALT,AST,ALP.The changes of serum BUN,UA,T-Bil and UCr.the changes of Hyp content in liver and kidney tissues.the pathological changes of liver and kidney tissues,α-smooth muscle actin（α-SMA）,transforming growth factor-β₁（TGF-β₁）and E-cadherin in liver and kidney tissues were observed.The results showed that after 4 weeks of induction,the liver and kidney function of rats was impaired,inflammatory factors were infiltrated,collagen deposition and fibrous tissue hypeiplasia in the liver and kidney tissues,indicating that the composite model of liver and kidney fibrosis was successfully established.In the third chapter of this paper,TAA combined with UUO method was used to induce liver and kidney fibrosis model in SD rats.The experimental period was 4 weeks.The expression of TNF-α in liver and kidney tissues,the deposition of type Ⅰ and Ⅲ collagen in liver and kidney tissues,and the expression of TLR4/MyD88/NF-κB pathway proteins were observed.The results showed that there were a large number of inflammatory factors in liver and kidney tissues during the modeling period.Meanwhile,TLR4/MyD88/NF-κB pathway protein expression was significantly up-regulated.In the fourth chapter of this paper,CCl₄ combined with adenine was used to induce the composite model of liver and kidney fibrosis in SD rats.The experimental period was 6 weeks.The body weight of model group and blank control group and survival rate were observed within 6 weeks.Blood samples were collected from orbit at 6 weeks after anesthesia.The changes of ALP,BUN,UA,T-Bil and Cr-were observed;the changes of Hyp content in liver and kidney tissues were observed;the pathological changes,α-smooth muscle actin（α-SMA）,transforming growth factor-β₁（TGF-β₁）and E-cadherin in liver and kidney tissues were observed.The results showed that inflammatory factors infiltration,collagen deposition,fibrous tissue proliferation,liver and kidney fibrosis were successfully established.In the fifth chapter of this paper,CCl₄ combined with adenine was used to induce the composit model of liver and kidney fibrosis in SD rats.The experimental period was 6 weeks.The expression of inflammatory factor TNF-α in liver and kidney tissues,the deposition of type Ⅰ and type Ⅲ collagen in liver and kidney tissues,and the expression of TLR4/MyD88/NF-κB pathway related proteins were observed.The results showed that there were a large number of inflammatory factors in liver and kidney tissues during the modeling period.Meanwhile,the expression of TLR4/MyD88/NF-κB pathway protein was significantly up-regulated.In this paper,TAA combined with UUO and CCl₄ combined with adenine were used to establish a composite model of liver and kidney fibrosis in rats.The results showed that TAA combined with UUO and CCl₄ combined with adenine could successfully establish a complex model of liver and kidney fibrosis in rats.The liver and kidney tissues induced by TAA combined with UUO showed infiltration of inflammatory factors.The survival rate was 65.52%after 4 weeks.The liver and kidney tissues of model rats induced by CCl₄ combined with adenine showed vacuolation of liver and kidney tissue cells,infiltration of inflammatory factors,diffuse fibrosis of intercellular tissue,vacuolation of liver and kidney tissue cells.The cycle was 6 weeks,and the survival rate of the rats was 80%.The modeling mechanism of TAA combined with UUO and CCl₄ combined with adenine may be due to the increased expression of TGF-β₁ and α-SMA,recruitment of inflammatory factors,activation of hepatic stellate cells,conversion of renal epithelial-mesenchymal to endothelium.Overexpression of the TLR4/MyD88/NK-κB signaling pathway,amplifies the cascade of inflammation and injury of liver and kidney.