马钱子碱诱导人单核细胞白血病THP-1细胞凋亡及作用机制  

Inducing-apoptosis Effect of Brucine on Human Monocytic Leukemia Cell Line THP-1 and Its Mechanism

作　　者：辛菲[1] 魏武[2] 纪爱芳[2] 申徐良[2] 张国香[2] 张梅香[2] 李仙仙[1] 张海燕[1] 

机构地区：[1]山西医科大学临床一系,山西太原030001 [2]长治医学院附属和平医院血液内科,山西长治046000

出　　处：《中国实验血液学杂志》2014年第3期681-686,共6页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目(30950015);山西省留学回国人员重点科研资助项目(2009-5)

摘　　要：本研究旨在探讨马钱子碱对人单核细胞白血病THP-1细胞的诱导凋亡作用及其可能的作用机制。应用CCK-8法检测马钱子碱对THP-1细胞的生长抑制作用;应用AO/EB荧光染色法、Annexin-Ⅴ/PI双染法检测马钱子碱对THP-1细胞凋亡的影响;PI单染法检测马钱子碱对THP-1细胞周期的影响;RT-PCR检测BCL-2、BAX基因的表达。结果表明,马钱子碱能有效地抑制THP-1细胞的生长,呈剂量和时间依赖关系;THP-1细胞经浓度为400μg/ml马钱子碱作用48 h后,大部分细胞核染色质被EB染成橘红色并呈固缩状,显示为晚期凋亡的细胞形态;0-400μg/ml范围内的马钱子碱作用THP-1细胞48 h后,随着药物浓度的增高,细胞凋亡率逐渐上升;与对照组相比细胞周期被阻滞于G0/G1期,并出现亚二倍体凋亡峰;RT-PCR结果显示,BCL-2基因表达明显下降,BAX基因表达上调。结论:马钱子碱能抑制THP-1细胞生长,诱导其凋亡,并对细胞周期起阻滞作用,其作用机制可能与BCL-2基因表达抑制及BAX基因表达激活有关。This study was aimed to investigate the inducing-apoptosis effect of brucine on human monocytic leukemia cell line THP-1 cells and its possible mechanism. The inhibition effect of brucine on growth of THP-1 cells was measured by CCK-8 method. Morphological changes of THP-1 cells treated with brucine was detected by acridine orange/ethidium bromide（ AO/EB ）double staining. Annexin-V/PI double labeling method was used to assay the apoptosis rate of THP-1 cells. The effect of brucine on THP-1 cell cycle distribution was detected by PI single staining. RT-PCR was used to detect the expression of BCL-2 and BAX. The results showed that the brucine could inhibit the THP-1 cell growth in concentration and time-dependent manners at the range of 50 to 400 μg/ml. The cells stained with AO/EB revealed that the brucine induced the nuclear chromatin condensation. After the THP-1 cells were treated with brucine of 400μg/ml for 48 hours, most nucleis were stained as orange-red, and condensed, displaying the late apoptotic celt morphology. Annexin-V/PI detection showed that brucine could induce apoptosis of THP-1 cells in a concentration-dependent manner. Compared with the control group, more cells in brucine-treated group were arrested at G0/G1 phase in a concentration-dependent manner. RT-PCR detection revealed that the expression of BCL-2 was down-regulated strikingly and BAX was up-regulated. It is concluded that brucine can efficiently inhibit cell growth and block THP-1 cells in Go/ G1 phase. The mechanism of THP-1 cell apoptosis induced by brucine may be related to the inhibition of BCL-2 and activation of BAX.

关 键 词：单核细胞白血病 THP-1细胞 马钱子碱 细胞凋亡 细胞周期 BCL-2基因 BAX基因 

分 类 号：R733.7[医药卫生—肿瘤]