| Objective:By establishing the model of carbon tetrachloride-induced liver fibrosis in mice,the improving effect of Yu Jin Pulvis(YJP)on liver fibrosis and its possible regulation mechanisms were explored,which could provide a powerful experimental basis for the development of new drugs.Methods:1.C57BL/6 mice were randomly separated into eight groups:(1)normal group(Normal,n=5),(2)CCl4 model group(CCl4,n=10),(3)group of low-dose YJP(CCl4+YJP-100 mg/kg,n=9),(4)group of medium-dose YJP(CCl4+YJP-200 mg/kg,n=9),(5)group of high-dose YJP(CCl4+YJP-300 mg/kg,n=10),(6)alone administration group of YJP(YJP-300mg/kg,n=9),(7)positive control group of Silybin(CCl4+Silybin-100 mg/kg,n=9),(8)positive control group of Fuzheng Huayu capsule(CCl4+FZHY-2 g/kg,n=9).Liver fibrosis in mice was established using 10%CCl4 via intraperitoneal injection for six weeks.The mice were administered with the remaining drugs once a day for six weeks.At the end of the sixth week,the mice were sacrificed to take their eyeball blood and liver tissue,and their general state was observed.2.Serum levels of ALT,AST,TNF-?,IL-1?,IL-12 and IL-18 were detected by ELISA.3.The color and smooth degree of mice liver were observed with naked eye.The pathological changes in liver tissues were evaluated by hematoxylin and eosin(H&E)and Sirius red staining.4.The positive expression of a-SMA and Collagen-I were measured by immunohistochemistry.The protein expression levels of a-SMA and Collagen-I were detected by Western blotting.The mRNA expression levels of a-SMA and Collagen-I were measured by reverse transcription polymerase chain reaction(RT-PCR).5.The protein expression levels of p-ERK/ERK,p-JNK/JNK and p-P38MAPK/P38MAPK in the MAPK pathway and p-PI3K/PI3K,p-Akt/Akt in the PI3K/Akt pathway were detected by Western blotting.Results:1.The mice in the normal group were in good condition,and there was no obvious abnormality in behavior,movement,food and water intake.The mice in the model group presented with lusterless hair and irritability.The above situation of mice in other groups was improved after treatment with drugs.2.Compared with the model group,the YJP and positive control groups could attenuate the levels of ALT,AST,TNF-a,IL-1?,IL-12 and IL-18 in serum.3.Anatomically,the livers were dark red and smooth surface in the normal group.There was a grainy appearance on the surface of liver tissue in the model group.Compared with the model group,YJP and positive control groups could improve the rough surface of liver tissue.The results of the two stainings consistently showed that the liver tissue of the model group had massive steatosis,inflammatory cell infiltration and collagen deposition.YJP could alleviate the above histological changes.4.The results of immunohistochemistry,Western blotting and RT-PCR were consistent.YJP could significantly decrease the expression levels of a-SMA and Collagen-I in liver tissue than that in the model group.5.Compared with that in the normal group,the model group significantly increased the phosphorylation of ERK,JNK,P38MAPK,PI3K and Akt.YJP and Silybin could significantly reduce the expression of p-ERK,p-JNK,p-P38,p-PI3K and p-Akt compared with the model group.However,Fuzheng Huayu capsule could only reduce the phosphorylation of JNK,P38,PI3K and Akt.Compared with the two positive control groups,YJP decreased the levels of p-ERK,p-P38 and p-Akt more significantly.Interestingly,the expression of p-Akt and p-P38MAPK were inhibited when treated with YJP alone rather than p-ERK,p-JNK and p-PI3K.Conclusions:1.Yu Jin Pulvis may alleviate the liver fibrosis by reducing the production of proinflammatory cytokines and extracellular matrix.2.Yu Jin Pulvis can also improve liver fibrosis by inhibiting the phosphorylation of MAPK signaling pathway families and PI3K/Akt at the same time.3.Yu Jin Pulvis has the value of anti-hepatic fibrosis. |
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