中药地椒乙醇提取物对人白血病细胞增殖抑制作用.docVIP

中药地椒乙醇提取物对人白血病细胞增殖抑制作用 　 作者：孙震晓，张英慧，程霜，马清温，郭善利，张金保 【关键词】 乙醇提取物 ［摘要］目的：筛选中药地椒乙醇提取物的抗肿瘤活性成分，探讨其可能的抗肿瘤作用机制。方法：从野生地椒乙醇提取物中分离得到乙酸乙酯、正丁醇和丙酮3种萃取组分，以体外培养的人白血病细胞株K562和HL60作为研究对象。将各萃取组分与肿瘤细胞作用一定时间后，计数存活肿瘤细胞数，分析药物对肿瘤细胞生长曲线的影响，考察各组分对肿瘤细胞生长的抑制作用。以去甲斑蝥素作为阳性对照组，通过形态学分析等方法，研究中药地椒乙醇提取物可能的抗肿瘤作用机制。结果：中药地椒乙酸乙酯萃取物对人白血病细胞株K562和HL60均有显著的抑制作用，且呈药物浓度依赖性，并可诱导肿瘤细胞凋亡；而正丁醇和丙酮萃取物对两种白血病细胞均无显著的抑制作用。结论：体外实验表明，乙酸乙酯萃取物是中药地椒乙醇提取物中主要的抗肿瘤活性成分。 　　［关键词］地椒; 乙醇提取物; 乙酸乙酯萃取物; 抗肿瘤药; K562; HL60 　　　Antitumor effect of ethanol extracts from Thymus quinquecostatus Celak on human leukemia cell line 　　ABSTRACT Objective: To screen the antitumor fraction of ethanol extracts from Thymus quinquecostatus Celak and investigate its antitumor effect on human leukemia cell line. Methods: Ethyl acetate, nbutanol and acetone fractions were separated from the ethanol extracts of wild Thymus quinquecostatus Celak. Growth inhibiting effects of these extracts on human leukemia cell lines K562 and HL60 were determined by live cell counting and cell growth curve analysis. The possible antitumor mechanism was studied by morphological analysis with norcantharidin as a positive control. Results: Ethyl acetate fraction could significantly inhibit the proliferations of K562 and HL60 cells, and the inhibiting effect depended on the concentration of ethyl acetate fraction. Ethyl acetate fraction could induce apoptosis of K562 and HL60 cells. The nbutanol and acetone fractions had no significant inhibiting effect on K562 and HL60 cells. Conclusion: Ethyl acetate fraction is the major antitumor fraction in ethanol extracts from Thymus quinquecostatus Celak. 　　KEY WORDS Thymus quinquecostatus Celak; ethanol extract; ethyl acetate fraction; antineoplastic agents; K562; HL60 　　中药地椒（Thymus quinquecostatus Celak），又名百里香，为唇形科百里香属植物，属矮小半灌木，茎匍匐生长，常生长于山坡和海边低丘上［1,2］。药用其地上部分，阴干或鲜用，味辛、性温，有小毒，有祛风解表、行气止痛之功［3］。民间单用地椒或配伍其他中药治疗癌症，确有一定的疗效。我们在对地椒进行有效成分提取、分离及分析的基础上［4,5］，以小鼠移植性肿瘤为模型，研究了地椒的抗肿瘤作用及其对机体免疫功能的影响［6］。随后，我们又以体外培养的人红白血病K562细胞和人早幼粒白血病HL60细胞为研究对象，进一步筛选地椒乙醇