痰热清注射液抑制白血病细胞体外增殖及其作用机制(英文)  

Inhibitory effects of traditional Chinese herbal medicine Tanreqing injection on proliferation of leukemia cells in vitro and the potential mechanisms

作　　者：杨波[1] 卢学春[1] 张峰[2] 刘洋[1] 朱宏丽[1] 

机构地区：[1]解放军总医院老年血液科,北京100853 [2]中国科学院北京基因组研究所,北京100029

出　　处：《中西医结合学报》2011年第4期414-422,共9页Journal of Chinese Integrative Medicine

基　　金：supported by the National Natural Science Foundation of China(No.30772597)~~

摘　　要：目的:探讨痰热清注射液对人白血病细胞体外增殖的影响及其作用机制。方法:将痰热清注射液以体积比稀释成1:2、1:4、1:8、1:16、1:32、1:64、1:128、1:256和1?512,共9个浓度组,分别处理增殖期慢性髓系白血病K562细胞和急性T淋巴细胞白血病Molt4细胞,镜下观察不同时间点各浓度组细胞生长情况,应用活细胞计数试剂盒检测细胞增殖能力,绘制生长曲线,计算抑制率和半数抑制浓度(half inhibitory concentration,IC50);分别采用碘化丙啶染色法和碘化丙啶/膜联蛋白V双染法,通过流式细胞术检测Molt4细胞周期及凋亡的变化;采用实时定量聚合酶链反应方法分析痰热清注射液处理Molt4细胞后凋亡相关基因caspase-3和bcl-2的表达量变化。结果:痰热清注射液对K562和Molt4细胞增殖具有抑制作用,1?2至1?16稀释浓度组的细胞毒性大,细胞基本死亡;抑制作用呈剂量和时间依赖性,IC50分别为1?333和1?142稀释浓度。痰热清注射液1?32浓度组处理72h后,Molt4细胞处于S期的数量明显减少(P<0.05),凋亡细胞比例明显增加(P<0.05);同时,caspase-3表达增加,bcl-2表达明显减少(P<0.05)。结论:痰热清注射液可抑制白血病细胞体外增殖并促进其凋亡,该作用可能是通过减少S期细胞数量,下调bcl-2基因表达和上调caspase-3基因表达而实现的。Objective：The present study investigates the effects of Tanreqing injection,a compound Chinese herbal medicine,on the proliferation of leukemia cells in vitro and discusses the potential mechanisms.Methods：Tanreqing injection was diluted to a series of concentrations（1︰2,1︰4,1︰8,1︰16,1︰32,1︰64,1︰128,1︰256 and 1︰512） by volume and then independently applied to treat chronic myeloid leukemia K562 cells and T cell acute lymphocytic leukemia Molt4 cells at the proliferative stage.Cell growth was observed at different time intervals under a microscope.Cell proliferation was determined by cell counting kit-8 assay and the survival curve was delineated.The inhibitory rate and the half inhibitory concentration（IC50） were calculated.Molt4 cells were stained with propidium iodide（PI） and PI/Annexin V and then the cell cycle and apoptosis were analyzed by using flow cytometry.In addition,a real-time quantitative polymerase chain reaction was subjected to detect the expressions of apoptosis-related genes（bcl-2 and caspase-3） after Tanreqing treatment.Results：Tanreqing injection had inhibitory effects on the proliferation of K562 cells and Molt4 cells.The most toxic concentrations were observed between 1︰2 and 1︰16 where cells were almost necrotic.The inhibitory effect manifested in a concentration-and time-dependent manner.The IC50 of K562 and Molt4 was 1︰333 and 1︰142,respectively.After 1︰32 Tanreqing injection treatment for 72 h,the number of Molt4 cells in the S phase significantly decreased（P0.05）,and the apoptosis rate markedly increased（P0.05）.In addition,increased caspase-3 expression and decreased bcl-2 expression were also observed（P0.05）.Conclusion：Tanreqing injection can both inhibit the proliferation and promote the apoptosis of leukemia cells in vitro,whereby the potential mechanism seems to be mediated in part by decreasing S phase ratio,down-regulating bcl-2 expression and up-regulating caspase-3 expression.

关 键 词：痰热清注射液 白血病 K562细胞 Molt4细胞 细胞增殖 细胞凋亡 

分 类 号：R733.7[医药卫生—肿瘤]