Effect of Xiayuxue Tang with Different Processing Method on CCl4-induced Hepatic Fibrosis in Rats / 中国实验方剂学杂志

ABSTRACT

Objective:To compare the anti-hepatic fibrosis effect of ethanol extract of honey Xiayuxue Tang (H-Xiayuxue Tang)and prepared slices of crude drug-Xiayuxue Tang (C-Xiayuxue Tang). Method:Totally 40 wistar rats were randomlydivided into normal group, model group, H-Xiayuxue Tang group, C-Xiayuxue Tang group, and Fuzheng Huayu group. Liver fibrosis model was induced by hypodermic injection of 50% carbon tetrachloride for 9 weeks. From the seventh week, the normal group and the model group were given 0.3% sodium carboxymethyl cellulose by gavage. The other three groups were given H-Xiayuxue Tang(0.58 g·kg-1), C-Xiayuxue Tang (0.43 g·kg-1), Fuzheng Huayu (2 g·kg-1), respectively. At the end of the ninth week, liver tissues and serum samples were collected for follow-up studies. The pathological changes and fibrosis degree were observed by HE and Sirius red staining, hydroxyproline content in liver tissues was detected by alkaline hydrolysis, serum liver function was determined by automatic analysis, the expressions of collagen type I (COL-I) and α-smooth muscle actin (α-SMA) in liver tissues were detected by Real-time PCR. The expressions of α-SMA in liver tissues was detected by Western blot and Immunohistochemistry, and serum inflammatory factors and oxidantion indexes were determined by ELISA, respectively. Result:Compared with control group, the severity of liver fibrosis was observed, the area ratio of Sirius red positive staining, hydroxyproline (Hyp) content, the protein and mRNA expressions of α-SMA, COL-I in the liver tissue were significantly increased in the model group (P<0.01). And the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (TBil), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-17A (IL-17A) and malondialdehyde (MDA) in the serum of the rats were markedly increased (P<0.01). The levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were apparently decreased (P<0.01). Compared with model group, the degree of liver fibrosis was significantly improved, the area ratio of Sirius red positive staining, the content of Hyp, the protein and mRNA expression of α-SMA and COL-I, and the serum levels of AST, ALT, TBil, TNF-α, IL-1β, IL-17A and MDA were significantly decreased (P<0.05, P<0.01). The serum contents of SOD and GSH-Px were markedly increased (P<0.05, P<0.01), respectively, by H-Xiayuxue Tang or C-Xiayuxue Tang. Among them, the improvement of liver histopathological changes was more obvious in the H-Xiayuxue Tang group than in the C-Xiayuxue Tang group. Meanwhile, the percentage of Sirius red positive staining area, the content of hydroxyproline, the expressions of COL-I and α-SMA mRNA and protein in liver tissue were significantly decreased compared with the C-Xiayuxue Tang group(P<0.05, P<0.01). In addition, there was no significant difference in serum liver function, inflammatory factors and oxidantion related indicators between groups, but the improvement trend of H-Xiayuxue Tang was more obvious. Conclusion:H-Xiayuxue Tang and C-Xiayuxue Tang have a better effect in hepatic fibrosis, especially H-Xiayuxue Tang, suggesting that honey can increase the anti-hepatic fibrosis effect of Xiayuxue Tang.