青蒿琥酯及神经酰胺对肝星状细胞增殖、胶原生成及MMP13表达的影响  

Effects of artesunate and ceramide on cell proliferation,collagen generated and MMP13 expression in hepatic stellate cells

作　　者：李英娴[1] 娄婷婷[1] 刘文东[1] 方步武[1] 

机构地区：[1]天津医科大学药理学教研室,天津300070

出　　处：《天津医科大学学报》2013年第4期271-274,共4页Journal of Tianjin Medical University

基　　金：国家自然科学基金资助项目(30772856)

摘　　要：目的:探讨青蒿琥酯及神经酰胺对大鼠原代肝星状细胞(HSCs)增殖以及胶原生成/分泌的影响,从诱导基质金属蛋白酶13(MMP13)表达这一环节探讨其抗肝纤维化的机制。方法:分离大鼠原代HSCs,传代培养活化后,将其分为实验组和对照组,实验组以青蒿琥酯(终浓度为125、150、175、200、225μmol/L)、神经酰胺(终浓度为10μmol/L)作用24、48、72 h。四甲基偶氮唑盐(MTT)法检测细胞增殖率,酶消化法测定羟脯氨酸(Hyp)含量,Western blot法测定MMP13的表达。结果:与细胞对照组相比,青蒿琥酯各浓度组及神经酰胺组作用24、48、72 h的吸光度值均降低(P<0.01),且对HSCs的抑制作用呈剂量-效应和时间-效应关系;均能有效降低HSCs上清液中Hyp含量(P<0.01),且呈剂量-效应关系;均可促进MMP13的表达(P<0.01),且呈剂量-效应关系。结论:青蒿琥酯及神经酰胺抗肝纤维化的可能机制是抑制肝星状细胞的增殖,减少胶原的生成/分泌,促进MMP13的表达,从而促进胶原的降解,使细胞外基质减少,抑制纤维化发生。Objective： To investigate the impact of artesunate and ceramide on cell proliferation and collagen＇generated, and to analyze the underlying molecular mechanisms of anti-fibrogenic effects involving matrix metalloprotenase 13（MMP13） expression in hepatic stellate eells（HSCs）. Methods： Isolated, cultured, and activated, HSCs were divided into seven groups, including one control group, five artesunate-treated experimental groups with 125, 150, 175, 200 or 225 μmol/L and one ceramide-treated experimental group with 10 μmol/L for 24, 48 or 72 hours. The rate of cellular proliferation was measured using the 3-（4, 5-dime-thylthiazol-2-yl）-2, 5- diphenyherazolium bromide （MTT） assay, and the determination of hydroxyproline （Hyp） was acquired by the enzyme digestion method. MMP13 expression was evaluated by Western blotting. Results： The absorbance value in artesunate and ceramide group reduced significantly compared with that in the control group at different points （P〈0.01）. As the concentration increasing and time forwarding, the inhibition of proliferation chanced; Artesunate in different doses and ceramide could effectively reduce the content of Hyp （P〈0.01）, and had a dose-effect relationship; Artesunate in different doses and ceramide could promote MMP13 expression （P〈0.01）, and also presented a dose-effect relationship. Conclusion： Artesunate and ceramide have anti-hepatic fibrosis effect. The possible mechanism may be that they can inhibit the proliferation of HSCs, reduce the secretion of collagen, improve the expression of MMP13 in order to promote degradation of collagen and decrease extracellular matrix.

关 键 词：青蒿琥酯 神经酰胺 肝星状细胞 基质金属蛋白酶13 

分 类 号：R96[医药卫生—药理学]