柴胡、柴胡和黄芩配伍对肝细胞损伤体外作用  

Water-extraction from radix bupleuri and compatibility of radix bupleuri and scutellaria baicalensis georgi on carbon tetrachloride-injured hepatocyte in vitro

作　　者：李小菲[1] 李静[1] 王義雯 李敏[1] 

机构地区：[1]陕西中医学院,陕西咸阳712046

出　　处：《吉林中医药》2016年第2期176-180,共5页Jilin Journal of Chinese Medicine

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

摘　　要：目的探讨柴胡、柴胡和黄芩配伍水提液对人肝L-02细胞增殖及对L-02细胞四氯化碳损伤的保护作用及其机制。方法本实验通过体外培养L-02细胞,将柴胡、柴胡和黄芩配伍水提液分别作用于人正常肝细胞及用CCL4损伤后的L02细胞。采用MTT法(体外细胞毒试验)测定柴胡、柴胡和黄芩配伍水提液对L02细胞增殖的影响;采用免疫组化法测定肝细胞中NFKBp65及TLR4蛋白的表达。结果 MTT法发现柴胡水提液对L02细胞有促进增殖作用,同时发现柴胡和黄芩配伍水提液对L02细胞有明显的增殖作用,而且配伍后作用更稳定。免疫组化发现CCL4损伤后L-02细胞中NKFBp65和TLR4蛋白表达增高,柴胡、柴胡和黄芩配伍水提液对肝细胞中的NKFBp65和TLR4蛋白具有抑制作用。结论柴胡水提液对L02细胞有促进增殖作用,柴胡和黄芩配伍水提液对L02细胞增值作用更稳定。且柴胡及柴胡和黄芩配伍可以对CCL4引起的肝损伤起保护作用,其机制可能与下调体内TLR4-NFKB信号通路有关。Objective To explore the effects of water-extraction from radix bupleuri and compatibility of radix bupleuri and scutellaria baicalensis georgi on the proliferation of human L-02 hepatocyte and protection of carbon tetrachloride-injured L-02 hepatocyte and Mechanism. Methods L-02 hepatocyte was cultivated in vitro in this research.Water-extraction from radix bupleuri and compatibility of radix bupleuri and scutellaria baicalensis georgi were used on human normal hepatocyte and L-02 carbon tetrachloride-injured hepatocyte respectively. The effect of water-extraction from radix bupleuri and compatibility of radix bupleuri and scutellaria baicalensis georgi on L-02 hepatocyte proliferation was measured by MTT assay（ cytotoxicity test in vitro）; the expression of NFKBp65 and TLR4 proteins in hepatocytes were determined by immunohistochemistry. Results MTT assay found that water-extraction from radix bupleuri and saikosaponin-d promoted the proliferation of L-02 hepatocyte,and water-extraction from compatibility of radix bupleuri and scutellaria baicalensis georgi promoted the proliferation of L-02 hepatocyte significantly and more stably.The expression of NKFBp65 and TLR4 in L02 cells increased after CCL4 injury. The compatibility of NFKBp65 and TLR4 in liver cells was inhibited by immunohistochemistry and the compatibility of radix bupleuri and scutellaria baicalensis georgi. Conclusion Water-extraction from radix bupleuri and saikosaponin-d promotes the proliferation of L-02 hepatocyte,and water-extraction from compatibility of radix bupleuri and scutellaria baicalensis georgi facilitates the proliferation of L-02 hepatocyte more significantly. Moreover,radix bupleuri and compatibility of radix bupleuri and scutellaria baicalensis georgi can protect hepatocyte from CCL4. The mechanism may be related to the down-regulation of TLR4-NFKB signaling pathway in vivo.

关 键 词：柴胡 黄芩 水提液 L02细胞 CCL4 NKFBp65 TLR4 

分 类 号：R285.5[医药卫生—中药学]