茅莓总皂苷和高三尖杉酯碱、阿糖胞苷体外协同诱导白血病细胞凋亡的机制研究  

The Synergy Mechanism Study of Total Saponins of Rubus Parviflolius L. co-administered with Either Homoharringtonine or Cytosine Arabinoside Inducing Leukemia Cell Apoptosis in Vitro

作　　者：许晓峰[1] 程汝滨[2] 杨波[2] 高瑞兰[3] 张学进[1] 

机构地区：[1]浙江省中西医结合医院血液科,浙江杭州310003 [2]浙江中医药大学药学院,浙江杭州310053 [3]浙江省中医院血液病研究所,浙江杭州310006

出　　处：《中华中医药学刊》2015年第7期1659-1663,I0008,I0009,共7页Chinese Archives of Traditional Chinese Medicine

基　　金：浙江省中医药管理局项目(2011ZA081;2012ZB120;2013ZB095;2014ZB089)

摘　　要：目的:探讨茅莓总皂苷(TSRP)抑制K562和人原代白血病细胞及分别与化疗药高三尖杉酯碱(Hom)、阿糖胞苷(Ara-c)的协同作用,并探讨其协同作用的可能机制。方法:MTT法和PHA-LCM液相-半固体二步培养法检测不同浓度的TSRP对K562和人原代白血病细胞增殖及其与化疗药联用后的影响;观察TSRP分别和Hom、Ara-c联用的抑制作用,用药物相互作用指数评价药物协同作用;DAPI染色法观察一定浓度下TSRP协同化疗药诱导K562细胞凋亡;Western blot检测促凋亡蛋白Cleaved caspase-3、Cleaved caspase-9和Cleaved PARP以及抗凋亡蛋白Survivin的表达。结果:TSRP能够抑制K562和人原代白血病细胞的增殖,呈浓度依赖性,在一定浓度范围内TSRP和化疗药有显著的协同作用;DAPI染色法观察TSRP和化疗药协同较单用化疗药出现明显凋亡现象,诱导细胞发生凋亡;Western blot检测TSRP联合Hom较单用Hom显著提高了K562细胞内PARP、Cleaved caspase-3和Cleaved caspase-9的活性;TSRP联合Ara-c较单用Ara-c显著提高了K562细胞内PARP和Cleaved caspase-3的活性,但是对Cleaved caspase-9的活性影响差别不大;TSRP联合Hom或Arac对Survivin的活性影响均未见明显差别。结论:TSRP分别和化疗药Hom或Ara-c联用对K562和人原代白血病细胞有明显的增殖抑制和诱导凋亡作用,且主要是通过线粒体途径增强凋亡效应,从而增加化疗药物的敏感性,具有良好的临床应用前景。Objective：To investigate the anti- proliferative activity of total saponins of Rubus parviflolius L. （TSRP）a- gainst K562 cells and leukemic progenitor cell as well as the synergistic effects and possible mechanism of TSRP co - ad- ministered with either Homoharringtonine （Horn） or cytosine arabinoside （ Ara - C ） , two major chemotherapy drugs. Methods ： MTT and PHA - LCM liquid and semi - solid culture were used to determine the anti - proliferative activity of different concentrations of TSRP against K562 cells and leukemic progenitor cell as well as the synergistic effects of TSRP and a chemotherapy drug. The inhibitory effect of TSRP combined with Horn and Ara - c, respectively, on cell proliferation was observed, and the synergistic effects were evaluated with the coefficient of drug interaction. DAPI staining was used to determine the synergistic effects of TSRP co - administered with a chemotherapy drug on inducting apoptosis of K562 cells. Western blot was used to determine the expression of thepro - apoptotic proteins including cleaved caspase - 3, cleaved caspase - 9 and cleaved PARP, and the anti - apoptosis protein - Survivin. Results ： TSRP was able to inhibit the proliferation of K562 cells and leukemic progenitor cell, and the inhibitory effect increased with the increasing concentra- tion of RPTS. In a certain concentration range, TSRP and chemotherapy drugs showed significant synergistic effects. DAPI staining indicated that the co - administration of TSRP and a chemotherapy drug showed a more significant effect on indu- cing apoptosis of K562 cells than the administration of a chemotherapy drug alone. Western blot assay indicated that the combination of TSRP and Hom significantly enhanced the activity of PARP, cleaved caspase - 3 and cleaved caspase - 9 in K562 cells compared with the administration of Hom alone ; the combination of TSRP and Ara - c significantly enhanced the activity of PARP and cleaved caspase - 3 in K562 ceils compared with the administration of Ara - c alo

关 键 词：茅莓总皂苷 K562 人原代白血病细胞 协同 凋亡 

分 类 号：R285.5[医药卫生—中药学]