柴胡皂苷d-黄芩苷配伍对CCl4诱导大鼠HSC的TLR4-NF-κB通路作用研究  

Inhibitory effects of SaikosaponinD-Baicalin on activation of rat HSC induced by CCl_4

作　　者：李敏[1] 高凯 王华林[1] 王義雯 李静[1] 王斌[1] Li Min;Gao Kai;Wang Hualin;Li Jing;Wang Bin(College of Pharmacy of Shaanxi University of Chinese Medicine,Xianyang 712046)

机构地区：[1]陕西中医药大学药学院,咸阳712046

出　　处：《陕西中医》2018年第1期3-5,共3页Shaanxi Journal of Traditional Chinese Medicine

基　　金：陕西省科技厅项目(2013JK4023);陕西省中医药管理局项目(ZY06);陕西省教育厅项目(12JK1016)

摘　　要：目的:探讨柴胡皂苷d-黄芩苷(SaikosaponinD-Baicalin,SB)对CCl4诱导肝星状细胞(Hepatic stellate cell,HSC)的作用机制。方法:HSC分空白对照(无药物处理),CCl4诱导(6mmol/L CCl_4作用6h),给药(6mmol/L CCl4作用6h后,2.5、5、10ug/ml的SB作用24h),TAK-242、BYA 11-7082组(6mmol/L CCl_4作用6h后,TAK-242 10mol/L、BYA 11-7082 2mol/L作用24h)。ELISA法检测透明质酸酶(Hyaluronidase,HA)、层粘连蛋白(Laminin,LN)、III型前胶原(ProcollagenⅢ,PCIII)、IV型胶原(Collagen Type IV,IV-C)浓度,RT-PCR、Western blot方法检测细胞中Toll样受体4(Toll-like receptors4,TLR4)、核转录因子(NF-κB)基因、蛋白的表达。结果:与CCl4干预组比较,SB、TAK-242、BYA 11-7082可降低CCl4造成的HSC细胞增殖,HA、LN、PCIII、HA细胞释放水平及NF-κB基因、蛋白的表达。SB、TAK-242可下调TLR4基因、蛋白表达。结论:SB可以抑制CCl4诱导的炎症因子基因、蛋白表达,改善肝纤维化相关指标,其作用机制可能与TLR4-NF-κB转录活性及蛋白活性相关。Objective:To investigate the inhibitory effect and mechanism of Saikosaponin D-Baicalin on activation of rat HSC induced by CCl4.Methods:The cells were divided into the following groups:control group(no treatment),model group(treated with6mmol/L CCl4treated for24h),SB groups(pretreated with6mmol/L CCl4treated for24h and2.5,5,10μg/ml SB for24h).Concentrations of HA,LN,PCIII,IV C were assayed by ELISA kits.Gene,protein expression of TLR4and NFκB were examined by RT-PCR and western blot analysis.Results:SB,TAK-242and BYA11-7082could significantly inhibit cell proliferation induced by CCl4.Furthermore they can downregulated gene and protein expressions of NFκB and the secretion of HA,LN,PCIII,IV C(P<0.001).Further studies disclosed that the SB and TAK-242could significantly inhibit the gene and protein expressions of TLR4.Conclusion:SB could inhibit inflammatory responses in HSC induced by CCL4probably by inhibiting the transcription activity and protein expression of TLR4-NF-κB which indicates its possible therapeutic effect in liver fibrosis.

关 键 词：肝纤维化 柴胡皂苷D-黄芩苷 肝星形细胞 CCL4诱导 TLR4 NF-κB 

分 类 号：R575[医药卫生—消化系统]