Study On The Prevention And Treatment Of Alcoholic Liver Fibrosis By Tiaoganlipi Prescription Based On TGF-?1/Smads Pathway And Its Related Mechanism

Objective:In this study,the rat model of alcoholic liver fibrosis was taken as the research object.this study observed the effect of Tiaoganlipi prescription on cytokines related to TGF-?1/Smads signaling pathway.In order to explore the prevention and treatment effect of Tiaoganlipi prescription on alcoholic liver fibrosis and its related mechanismMethods:Alcohol intragastric method was used to establish rat model of alcoholic liver fibrosis.112 SD rats were randomly divided into 7 groups after adaptive feeding for 3 days.They were set as blank control group(Con,10),model group(AHF,17),low dose group(L,17),medium dose group(M,17),high dose group(H,17),prevention group(P,17)and control group(F,17).Except for the Control group,Rats in model group d were given 56° liquor by gavage.Rats in model group were given 5 6° liquor by gavage.Diluted by concentration gradient of 40%,4 5%,50%and 56%,respectively.The concentration of alcohol increases once every four weeks.The adaptability of gavage volume increased by 0.5?1ml/100g.At week 1 to 4,4 0%alcohol 4g/(kg·d)was administered by gavage;At 5?8 weeks,4 5%alcohol was given 4.5g/(kg·d);At week 9-12,50%alcohol 5g/(kg·d)was given;At week 13 to 18,56%alcohol was given 5.6g/(kg·d).At the end of the 18th week of alcohol intragastric modeling,the dosage of Tiaolilipi prescription was given to group L(0.5 g/kg),group M(1 g/kg),group H(2g/kg),and group F(0.54g/kg)respectively.At the end of the 22nd week,the rats were sacrificed and the liver tissue morphology was observed.At the end of the 22nd week,the rats were sacrificed and the liver tissue morphology was observed.After HE staining and Masson trichrome collagen staining,the pathological changes of the liver were observed under the microscope.The serum levels of ALT,AST,HA,CIV and LN,as well as the contents of TGF?-1,TNF-?,IL-1 and IL-10 in liver tissues were determined by ELISA.The expression levels of TGF?-1,Smad2/3 and Smad7 were detected by PT-PCRResults:(1)The changes of general indexes of rats were as follows.Compared with the control group,the body weight of AHF group decreased significantly(P<0.05)and the liver and spleen index increased(P<0.05,P<0.01).Compared with the AHF group,the weight of rats increased significantly(P<0.05)and the index of liver and spleen decreased(P<0.05,P<0.01)after the administration of Tiaoganlipi prescription.However,the difference was not statistically significant in the low dose group(P>0.05).Observe the texture,color and morphology of the liver with the naked eye.The appearance of the liver in group M,group H,group P and group F was close to normal(2)The pathological changes are as follows:In the AHF group,HE staining showed inflammatory cell infiltration,hepatic lobules and focal necrosis,and fibrous connective tissue hyperplasia.After the administration of Tiaoganlipi prescription,the degree of inflammatory cell infiltration and the hyperplasia of fibrous connective tissue were significantly reduced.Transmission electron microscope,compared with Con group,AHF group of liver cell organelles and the decrease in the number of mitochondria,rough endoplasmic reticulum,part has divided period or a spindle into muscle cells,extracellular collagen fibers increases,vacuoles in cells reduced in different degrees,hepatic stellate cells(hepaic stellate cells,HSCs)within the lipid drops,did not see obvious proliferation activity,a small amount of collagen fiber hyperplasia;Observations of Masson staining.In the AHF group,there were significantly more fibers in the liver tissue and the hepatic lobular structure was destroyed;A large number of blue collagen fibers surround the portal vein area;Fiber spacing increases and widens.After the treatment group with Tiaoganlipi prescription,the hepatic lobular structure was slightly damaged and the collagen fibers became thinner and less spaced.Collagen deposition was significantly reduced.The score of hepatocyte degeneration was decreased(3)Compared with the control group,the contents of AST,ALT,HA,LN and CIV in serum of rats in AHF group were significantly increased(P<0.01).Compared with the AHF group,the contents of AST,ALT,HA,LN and CIV were decreased to different degrees(P<0.05,P<0.0 1),after the intervention of Tiaoganlipi prescription(4)Compared with the control group,the contents of TGF?-1,TNF-?,and IL-1 were significantly increased in the AHF group(P<0.01),and the contents of IL-10 were significantly decreased(P<0.05).Compared with the AHF group,after the intervention of Tiaoganlipi prescription,the contents of TGF ?-1,TNF-?,and IL-1 were significantly reduced(P<0.05,P<0.0 1)and the contents of IL-10 significantly increased(P<0.05)(5)Compared with the control group,the expression levels of TGF?-1?Smad2 and Smad3 mRNA in the liver tissues of AHF group were significantly increased(P<0.01)and the expression levels of Smad7 mRNA significantly decreased(P<0.01).Compared with the AHF group,the expression of TGF?-1,Smad2 and Smad3 mRNA decreased significantly(P<0.01)and the expression of Smad7 mRNA increased significantly(P<0.0 5,P<0.0 1).Among them,the high dose of Tiaoganlipi prescription and prevention group had the best therapeutic effectConclusion:1.The AHF model could be successfully established by intragastric administration of SD rats with different concentrations of alcohol for 1 8 weeks2.The results of weight,liver and spleen index,HE staining,transmission electron microscopy,Masson staining,liver function and liver fiber indexes of rats in the AHF group showed that Tiaoganlipi prescription could significantly improve the degree of liver injury and liver fibrosis in rats3.Tiaoganlipi prescription can reduce the contents of IL-1,TNF-? and TGF?-1 in the liver tissues of rats in the AHF group,increase the content of IL-10,reduce the generation of ECM and its effect on the downstream TGF?-1/Smads pathway.Therefore,the mechanism of Tiaoganlipi prescription to reduce alcohol-induced fibrosis in rats may be related to the inhibition of the expression of IL-1,IL-10,TNF-? and TGF?-14.Tiaoganlipi prescription can downregulate the expression of TGF?-1,Smad2 and Smad3 mRNA,and up-regulate the expression level of Smad7 mRNA.The therapeutic and preventive effects of Tiaoganlipi prescription on alcoholic liver fibrosis may be realized by affecting the expression of TGF-livelihood 1/Smads signaling pathway related cytokines.