ABSTRACT
Objective::To investigate the relationship between Toll-like receptor 4 (TLR4) mRNA and its downstream inflammatory factor-tumor necrosis factor-α (TNF-α) and hepatocyte apoptosis in mice with intestinal endotoxemia (IETM) of hepatic failure, and explore the regulatory mechanism of Wenyang Jiedu Huayu granule on endotoxin-induced hepatocyte apoptosis. Method::The 85 SPF male SD rats were randomly divided into normal group, model group, TLR4 monoclonal antibody group and Wenyang Jiedu Huayu granule group. D-galactosamine (D-Gal) intraperitoneal injection was performed to establish the IETM model of hepatic failure. The TLR4 monoclonal antibody group and the Wenyang Jiedu Huayu granule group were given Wenyang Jiedu Huayu granule solution by gavage 5 days before the modeling. The normal group and the model group were given isovolumetric distilled water. Each group was given by gavage until sacrifice. Rats in each group were randomly sacrificed at 24, 48, 72 h, respectively, and samples were collected. The levels of serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected at 24, 48, 72 h. The pathological changes of liver tissue were observed by hematoxylin and eosin(HE) staining. The expression of TLR4 in liver tissue was detected by real-time fluorescence quantitative PCR (Real-time PCR). The expression of TNF-α in liver tissue was detected by enzyme-linked immunosorbent assay (ELISA). The apoptotic rate of hepatocytes was detected by flow cytometry. Result::Compared with the normal group, ALT and AST were increased in model group, while the pathological injury degree of liver tissue was significantly increased. The expressions of TLR4 mRNA and TNF-α were increased (P<0.05, P<0.01), whereas the apoptosis rate of liver cells was significantly increased (P<0.05, P<0.01). Compared with the model group, ALT and AST were decreased in Wenyang Jiedu Huayu granule group (P<0.01), and the degree of pathological injury of liver tissue was significantly reduced (P<0.05). The expressions of TLR4 mRNA and TNF-α were significantly decreased (P<0.01), and the apoptosis rate of liver cells was also reduced (P<0.01). Conclusion::TLR4 mRNA and TNF-α are positively correlated with hepatocyte apoptosis in liver failure. Wenyang Jiedu Huayu granule can improve liver function, alleviate liver cell injury and reduce liver cell apoptosis in IETM mice with hepatic failure. The mechanism may be related to its ability to down-regulate the expression of liver TLR4 mRNA, inhibit the release of TNF-α, and reduce the rate of hepatocyte apoptosis.