Exploration of the Effect and Mechanism of Shuangzhu Kangxian Prescription in Improving Carbon Tetrachloride-Induced Hepatitic Fibrosis in Rats Based on Wnt/β-catenin Signaling Pathway
Objective To investigate the effect and mechanism of Shuangzhu Kangxian Prescription(Astragali Radix,bran-fried Atractylodis Macrocephalae Rhizoma,vinegar-prepared Rhizoma Curcumae,Bupleuri Radix,Salviae Miltiorrhizae Radix et Rhizoma,Litchi Semen)on improving hepatitic fibrosis induced by carbon tetrachloride(CCl4)in rats based on the Wnt/β-catenin signaling pathway.Methods The rat model of hepatitic fibrosis was replicated by subcutaneous injection of 3.0 mL·kg-1 40%CCl4 twice a week for 8 weeks.SD rats were randomly divided into blank control group(n=8),model group(n=7),positive control group(n=7,intragastric administration of 43.19 mg·kg-1 silymarin),low-dose Chinese medicine group(n=6,intragastric administration of 4.3 g·kg-1Shuangzhu Kangxian Prescription),medium-dose Chinese medicine group(n=6,intragastric administration of 8.6 g·kg-1Shuangzhu Kangxian Prescription),high-dose Chinese medicine group(n=7,intragastric administration of 17.2 g·kg-1Shuangzhu Kangxian Prescription).The continuous intragastric administration was given once a day for 4 consective weeks,in addition to the blank control group,the other groups continued to be subcutaneously injected with CCl4 at the same time.The pathological changes of liver tissue were observed by HE and Masson staining.The levels of serum type Ⅳ collagen(Col Ⅳ),type Ⅲ procollagen(PC Ⅲ),hyaluronic acid(HA)and laminin(LN)were detected by ELISA.The protein expression levels of Wnt1,β-catenin and PPAR-γ in liver tissue were detected by Western Blot.The mRNA expression levels of Wnt1,β-catenin and PPAR-γ in liver tissue were detected by qPCR.Results Compared with the blank control group,the liver of the model group showed partial necrosis of liver cells,the normal hepatic lobule structure was destroyed,the arrangement of liver cell cords was disordered,the central vein or portal area was enlarged,and a large number of inflammatory cells infiltrated to form bridging necrosis.The collagen fibers in the central vein or portal area of the liver proliferated significantly,forming fibrous septa,and the fibrosis score were significantly increased(P＜0.05).The levels of serum Col IV,PC Ⅲ,HA and LN were significantly increased(P＜0.05).The protein and mRNA expression levels of Wnt1 and β-catenin in liver tissue were significantly increased(P＜0.05),and the protein and mRNA expression levels of PPAR-γ were significantly decreased(P＜0.05).Compared with the model group,the steatosis of hepatocytes in the positive control group and the medium-and high-dose groups of Chinese medicine was improved,and the necrosis and inflammatory cell infiltration were reduced.The degree of hepatitic fibrosis was improved,the liver collagen fibers were reduced,and the fibrosis score was significantly reduced(P＜0.05).The levels of serum Col Ⅳ,PC Ⅲ,HA and LN were significantly decreased(P＜0.05).The protein and mRNA expression levels of Wnt1 and β-catenin in liver tissue were significantly decreased(P＜0.05),and the protein and mRNA expression levels of PPAR-γ were significantly increased(P＜0.05).Conclusion Shuangzhu Kangxian Prescription has the effect of anti CCl4-induced hepatitic fibrosis in rats,and its mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway and up-regulation of PPAR-γ expression.
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