| Objective:Previous studies of this subject showed that Fugan Huaxian Dec-oction can improve serum index and pathological changes of liver tissue in r-ats with hepatic fibrosis(HF).On this basis,we combine the basic theory of Chinese medicine,this topic combines the classic theory of "hepatitis,spleen and cultivating spleen",and establishes the syndrome of "Zhengxu toxic and blood stasis" combined with liver fibrosis animal model,and to explore the effect o f Fugan Huaxian decoction on the liver fibrosis model rat with weakened bod y resistance,toxic accumulation and blood stasis,combining disease and syndro me from the perspective of MAPK signaling pathway regulation.Methods:120 SD rats of clean grade were fed for one week.Weighed and randomly divided into six groups,each group of 20,respectively,the normal co-n trol group(group A),model group(group B),colchicine treatment group(grou-p C),Fugan Huaxian decoction high dose group(D Group),Fugan Huaxian De-c oction medium dose group(E group),Fugan Huaxian Tang low dose group(F group).Except for the normal group,the other groups were guided by TCM th eory and used to build the liver fibrosis rat model of weakened body resis-ta nce,toxic accumulation and blood stasis combining disease and syndrome in a ddition to the classical subcutaneous injection of carbon tetrachloride.6 weeks after modeling,2 rats were randomly selected from each group and used for li ver tissue HE staining and Masson staining test model,and the intragastric do se was calculated according to the clinical equivalent amount conversion ra-te of adult to rat.Group C was intragastrically given colchicine(2mg·kg-1·d-1),an d group D was intragastrically given Fuganhuaxian decoction at a high dose.(67.08g·kg-1·d-1),group E was intragastrically given Fuganhuaxian Decoction at a medium dose(33.54g·kg-1·d-1),group F was intragastrically given Fuganhuaxi an Decoction at a low dose(16.77g·kg-·d-1),group A and B were given an eq-ual volume of purified water.The general condition of the rats was observed on the day following the last gavage.Control group(purified water 2ml/d),mod-el group(purified water 2ml/d),colchicine treatment group(2mg·kg-1·d-1),Fuga-n Huaxian Decoction high-dose treatment group(67.08g·kg-1·d-1),Fugan Huaxi an Decoction medium dose treatment group(33.54g·kg-1·d-1),Fugan Huaxian Decoction low dose treatment group(16.77g·kg-1·d-1).The general condition of the rats was observed on the day following the last gavage.The rats were sacr-ificed after fasting for 12 hours,and blood and liver tissues were frozen and stored in a-80?refrigerator for use.The aspartate aminotransferase(AST)and alanine aminotra-nsferase(ALT)was measured by microplate method,the albu-min(ALB)was measured by micro ELIASA method,and the content of hydr-oxyammonia(Hyp)was measured by alkaline hydrolysis method;The levels of type III pro-collagen peptide(PIIINP)and laminin(LN)were determined by ELISA.The degree of liver injury and pathology of the rats after treatment were observed under HE and Masson staining.The expression of ERK1/2,JNK 1/2 and P38 related proteins and their phosphorylated proteins in liver MAPK signaling pathway was detected by Western-Blot method.The results of quanti-tative analysis were P-ERK/ERK,P-JNK/JNK,P-P38.The ratio of P38 is based on the analysis of the effect and effect of Fugan Huaxian Decoction on HF production through MAPK signaling pathway.Results:This research task is intended to establish an liver fibrosis animal model of weakened body resistance,toxic accumlation and blood stasis,combi-ning disease and syndrome model success,The general condition of the rats in each treatment group was worse than that in the normal control group,but it was significantly better than the model group,but there was no significant dif-ference between the treatment groups.Except the model group,the body weig-ht of each group increased(P<0.0 lorP<0.05),but there was no significant dif-ference in body weight between the treatment groups.ALT,AST,ALB and live-r fibrosis four Hyp,LN,PIIINP,the treatment group and the model group were improved,among which the high dose group was the most obvious(P<0.05);Fugan Huaxian Decoction medium and low dose group and There was no sig-nificant difference between the colchicine groups(P>0.05).HE staining of live-r tissue pathological sections showed that compared with the model group,Fu-gan Huaxian Decoction had a small amount of collagen fiber deposition and inflammatory cell infiltration in the high dose group,and the hepatic cord wa-s arranged neatly and without pseudo-lobule formation;colchicine group and li-ver There were no significant differences bet-ween the medium and low dose groups of Huaxian Decoction.There were different degrees of collagen fibrosi-s and hepatocyte degeneration and necrosis.A few of them had incomplete ps-eudolobule formation,hepatic cord structure was disordered,and a large number of inflammatory cells infiltrated.Masson-stained liver histopathological sectio-ns showed that compared with the model group,the lobular structure of the co-lchicine group and the Fugan Huaxian Decoction in the middle and low dose groups were basically present,the portal area was enlarged,and the fibrous tis-sue was proliferated.The high-dose group of Fugan Huaxian Decoction has co-mplete lobular structure,a small amount of fibrous tissue hyperplasia,and the effect of improving liver fib-rosisis most obvious.Western-Blot results showed that P38,ERK1/2,and JNK1/2 were not statistically different among the group-s.P-P38,P-P38/P38,P-ERK1/2,P-ERK/ERK,P-JNK1/2,P-JNK/JNK ratio were stati-stically different between the normal control group and the model group and the treatment group.The expression of the model group was significantly hig-her than that of the treatment group.The highest expression of Fugan Huaxian Dec-oction in the treatment group was the lowest in the treatment group,com-pared with the middle,low and colchicine treatment groups(P<0.01 or P<0.05).There was no significant difference between the low-dose group of Fugan Hu axian Decoction and the colchicine,high-dose and medium-dose groups.Conclusion:1.Base on Subcutaneous injection and combination of CCL4 olive oil mixture,establish an liver fibrosis animal model of weakened body resistance,toxicc accumlation and blood stasis,combining disease and syndrome.2.Fugan Huaxian Decoction can reduce the model of Zhengxu toxic and bloo d stasis the serum(ALT,AST,HYP,LN,PIIINP)content of rats with liver fibrosi s and increase the content of ALB.3.Can reduce the degree of liver fibrosis i n rats;4.Fugan Huaxian Decoction can alleviate or re-gulate liver fibrosis by d own-regulating the expression of P-P38,P-ERK1/2 and P-JNK1/2. |
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