白花蛇舌草水提物抗白血病CEM细胞增殖作用及机制.docVIP

白花蛇舌草水提物抗白血病CEM细胞增殖作用及机制 　 作者：陈秀珍，朱大诚，王艳辉，欧阳永伟，尹小明 【摘要】 目的观察白花蛇舌草水提物对CEM细胞增殖影响及其机制。方法对CEM细胞进行细胞培养。采用台盼蓝拒染法，镜下计数活细胞数，绘制生长曲线，检测白花蛇舌草水提物对CEM细胞的生长抑制作用;琼脂糖凝胶电泳法检测白花蛇舌草水提物对CEM细胞的诱导凋亡作用。结果白花蛇舌草水提物终浓度0.64，3.2，16μg/ml均显著抑制CEM细胞的生长，且呈剂量和时间依赖性。琼脂糖凝胶电泳结果显示实验组出现明显的DNA梯形凋亡条带，而空白对照组没有出现梯形条带。结论白花蛇舌草水提物对CEM细胞生长具有显著抑制作用，诱导肿瘤细胞凋亡可能是其主要机制之一。 【关键词】 白花蛇舌草; CEM细胞; 生长抑制; 细胞凋亡 　　Abstract：ObjectiveTo observe the inhibitory function and the mechanisms of the aqueous extract of Hedyotis diffusa Willd on the CEM cells. MethodsCulture the CEM cells. Apply trypan blue exclusion to take count of the live cells in the microscope, protract the growth curve and detect the growth inhibitory function of aqueous extract of Hedyotis diffusa Willd on CEM cells. Agarose gel electrophoresis was used to detect the induced apoptosis effects of Hedyotis diffusa Willd aqueous extract on CEM cells.ResultsThe aqueous extract of Hedyotis diffusa Willd inhibited the growth of CEM cells obviously as the medicine concentration were 0.64μg/ml, 3.2μg/ml or 16μg/ml. The aqueous extract of Hedyotis diffusa Willd could induce apoptosis of CEM cells in dose and time-dependent manner. An obvious apoptotic DNA ladder appeared in the experimental group within the agarose gel electrophoresis, while in the control group was on contrary.ConclusionThe aqueous extract of Hedyotis diffusa Willd can inhibit the growth of CEM cell strongly. To induce apoptosis is one of its mechanisms. 　　Key words：Hedyotis diffusa Willd; CEM cell; Growth inhibition; Apoptosis 　　白花蛇舌草Hedyotis diffusa Willd为茜草科一年生草本植物白花蛇舌草的全草，其味微苦、微甘，性凉，入心、肝、肺经，具有清热、解毒、利湿、抗癌等功效。近年来许多学者对白花蛇舌草进行了广泛的研究，特别是其抗肿瘤作用的报道较多，但其对白血病作用的研究较少[1，2]。本实验研究白花蛇舌草水提物对白血病CEM细胞增殖影响，探讨其抗白血病的机制，为拓宽其在临床上的应用提供实验依据。 　　1 材料和方法 　　1.1 药品及试剂白花蛇舌草由江西药都樟树中药饮片有限公司生产，批号白花蛇舌草水提物的提取方法参照陈秀珍等[3]报道的方法进行，最终配成浓度为220 mg/ml的白花蛇舌草水提物备用。RPMI1640为Gibco公司产品，小牛血清(FBS)为杭州四季清生产，DNA Ladder检测试剂盒购于凯基生物科技发展有限公司。 　　1.2 细胞培养白血病CEM细胞株本实验室保存。培养基为含体积分数为10%的FBS的RPMI1640，并加入双抗(青霉素100 U/ml和链霉素100 μg/ml)。在37℃，5%CO2，饱和湿度的培