Inhibitory effects of Bupleurum chinense-Scutellaria baicalensisdecoction on activation of rat HSC induced by CCl4 / 中国药理学通报

Aim To investigate the inhibitory effect of Bupleurum chinense-Scutellaria baicalensis decoction(CQ)on activation of rat HSC induced by CCl4 and the mechanism.Methods Cells in logarithmic growth phase were cultured in culture medium without FBS for 24 h.After disassociated using 0.25％EDTA-trypsin,the cells were seeded into respective plates at the density of 1.5×109· L-1 and cultured overnight.The cells were divided into the following groups:control group(no treatment),model group(treated with 6 mmol·L-1 CCl4 for 24 h),CQ groups(pretreated with 6 mmol·L-1 CCl4 for 24 h and 400,500,600 mg·L-1 CQ for 24 h).Concentrations of hyaluronidase(HA),laminin(LN),procollagen Ⅲ(PCⅢ),collagen type Ⅳ(Ⅳ-C)were assayed by ELISA kits.Gene expressions of Toll-like receptor 4(TLR4)and NF-κB were examined by RT-PCR analysis.Protein expression of TLR4 and NF-κB was examined by Western blot.Results CQ could significantly inhibit cell proliferation induced by CCl4.Furthermore,CQ at 600 mg·L-1 significantly downregulated gene and protein expressions of TLR4 and NF-κB.And CQ reduced the secretion of HA,LN,PCⅢ,Ⅳ-C.Further studies disclosed that the TLR4 inhibitor TAK-242 and NF-κB inhibitor BAY 11-7082 could significantly inhibit the gene and protein expressions of NF-κB,but could not change gene and protein expression of TLR4,and reduced the secretion of HA,LN,PCⅢ,Ⅳ-C.Conclusion CQ could inhibit inflammatory responses in HSC induced by CCl4 probably by inhibiting the transcription activity and protein expression of TLR4-NF-κB,which indicates its possible therapeutic effect on liver fibrosis.

Protective Effect and Mechanism of Matrine Combined with Glycyrrhizic Acid in the Treatment of Chronic Liver Injury Induced by Carbon Tetrachloride / 中国药师

Objective: To investigate the protective effects of matrine combined with glycyrrhizic acid on chronic liver injury induced by carbon tetrachloride, and explore the protective mechanism from the points of energy metabolism and CYP enzyme.Methods: The chronic hepatic injury model of rats was induced by CCl4.The changes of activity of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were measured to observe the protective effect of the two drugs and their combination.The contents of glutamate dehydrogenase (GLDH) in serum and adenine nucleoside three phosphate (ATP), adenosine diphosphate (ADP) and adenine monophosphate (AMP) in liver tissue were determined to evaluate the regulation effect on hepatic energy metabolism and mitochondrial function.The levels of CYP1A2, CYP2E1 mRNA and protein in liver tissue were detected by real-time PCR and Western Blot to evaluate the two drugs and their combination on the regulation function of liver CYP enzyme.Results: Matrine (72.8 mg×kg-1)and glycyrrhizic acid(43.4 mg·kg-1)could decrease the serum activities of ALT and ALT in chronic hepatic injury model, and the combination (matrine 36.4 mg·kg-1+glycyrrhizic acid 21.7 mg·kg-1) had the most significant protective effect (P＜0.05).Matrine (72.8 mg·kg-1)and glycyrrhizic acid(43.4 mg·kg-1)could decrease GLDH in serum,and restore the content of ATP in liver (P＜0.05).Matrine (72.8 mg·kg-1) had no effect on the expression of CYP1A2 and CYP2E1mRNA, and glycyrrhizin (43.4 mg·kg-1) could inhibit the expression of CYP1A2, CYP2E1mRNA and protein (P＜0.05).Conclusion: Matrine combined with glycyrrhizin has obvious regulation effect on mitochondrial function and liver protective effect in chronic hepatic injury model.