甘草总黄酮与石见穿总酚酸联用促肝星状细胞凋亡作用.docVIP

　　甘草总黄酮与石见穿总酚酸联用促肝星状细胞凋亡作用 【摘要】 目的 研究甘草总黄酮与石见穿总酚酸联用对氧化应激的肝星状细胞(HSC)的促凋亡作用。方法 MTT法检测甘草总黄酮与石见穿总酚酸联用对大鼠肝星状细胞株(HSC-T6)增殖的影响。次氮基三乙酸铁与HSC-T6体外共培养建立氧化应激模型,采用TBA法检测细胞内丙二醛(MDA)含量,运用邻苯酸酚自氧化速率法检测超氧化物歧化酶(SOD)活性。荧光探针DCFH-DA检测细胞内活性氧水平。荧光染料Hoechst33258观察细胞的形态及凋亡情况。JC-1检测细胞内线粒体膜电位变化情况。结果 甘草总黄酮与石见穿总酚酸联合处理HSC-T6后,细胞的增殖受到抑制,并促进HSC-T6凋亡;细胞内线粒体膜电位下降;随着药物剂量的增加细胞内活性氧含量下降。结论 甘草总黄酮与石见穿总酚酸联用促进氧化应激的HSC凋亡,此作用可能与线粒体凋亡途径有关。 【关键词】 甘草总黄酮;石见穿总酚酸;肝星状细胞;凋亡;线粒体膜电位 　Abstract：Objective To investigate the effect of Glycyrrhiza uralensis flavonoids and salvianolic acid (GS) on the apoptosis of HSC-T6 cells. Methods HSC-T6 etric method and trypan blue staining easure the cell proliferentiation and survival rate. Production of reactive oxygen species (ROS) of drug-treated cells onitored by a fluorescent probe 2’,7’-dichlorodihydrofluorescin fluorescence (DCFH-DA). Morphological changes associated ined by staining itochondrial transmembrane potential inde by the dual-emission potential-sensitive probe 5,5,6,6-tetra-chloro-1,1,3,3-tetraethyl-imidacarbocyanine iodide (JC-1). Result Oxidative stress decreased the activity of SOD in HSCs, increased the content of MDA in HSC-T6. GS inhibited the cells proliferation and decreased the iROS levels in a dose dependent manner, After 24 hours treatment orphology ic ratio and the increase of kidney-shape nuclear cells. Furthermore, GS induced a concentration decrease in ROS production and in mitochondrial membrane potential. Conclusion GS play an important role in oxidative stress HSC-T6 apoptosis ay be corrected itochondria apoptosis pathitochondria membrane potential 　　肝纤维化(hepatic fibrosis,HF)是慢性肝病的共同病理学基础,其中25%～40%最终发展为肝硬化甚至肝癌 [1]。目前认为,肝星状细胞(hepatic stellate cells,HSC)是HF细胞外基质的主要细胞,HSC的活化和功能的改变是HF形成的关键[2-3],也是抗纤维化研究的核心。越来越多的实验表明,氧化应激是不同原因引起的HF发生的共同通路。实际上,铁负载、乙醇、四氯化碳和丙型肝炎病毒等引起HF的病因都可通过氧化应激和脂质过氧化途径激活HSC。Iredale等[4]的实验表明,激活的HSC主要不是通过转化为静止的HSC实现的,也并非引起坏死,因为纤维间隔及其邻近区域没有坏死及炎症的表现,而是凋亡的结果。在急性或慢性肝损伤修复的动物模型中也发现了HSC的凋亡。由激活的HSC分泌的金属蛋白酶组织抑制剂(TIMP)的表达减少,解除了对胶原酶活性的抑制使胶原降解增加,再加上HS