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

ABSTRACT

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 groupscontrol 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.