苦参碱诱导NB4细胞凋亡作用及机制  

Mechanism of matrine-induced apoptosis in NB4 cells

作　　者：李瑞[1] 朱玉良[1] 董晶[1] 张紫阳[1] 董均明 马玲[1] 

机构地区：[1]哈尔滨医科大学公共卫生学院,黑龙江哈尔滨150081

出　　处：《中国公共卫生》2016年第6期785-788,共4页Chinese Journal of Public Health

基　　金：黑龙江省教育厅项目(1154h20)

摘　　要：目的探讨苦参碱(Mat)诱导急性早幼粒白血病细胞株NB4细胞凋亡作用及机制。方法不同剂量M at(0.25、0.50、1.00 mg/m L)作用于NB4细胞(24、48、72 h),噻唑蓝法(M TT)检测细胞增殖抑制率;Hoechst33258染色观测细胞形态学变化;流式细胞术(FCM)检测细胞周期,Annexin V-FITC/PI双标记法测定细胞凋亡率;分光光度法检测NB4细胞内caspase-3和caspase-8活性。结果与对照组比较,Mat可明显抑制NB4细胞增殖,诱导细胞凋亡,呈时间和剂量效应(P<0.05);细胞形态学观察可见明显的细胞凋亡特征;与对照组比较,0.50、1.00 mg/m L M at组G0/G1期细胞比例(分别为57.91%、83.00%)明显增加,S期比例(分别为40.95%、16.10%)减少;与对照组比较,0.50、1.00 mg/m L Mat组NB4细胞内caspase-3酶活性[分别为(77.41±4.01)、(111.78±4.05)]、1.00 mg/m L M at组NB4细胞内caspase-8酶活性(44.98±7.63)明显增强(P<0.05)。结论 M at可抑制NB4细胞增殖并诱导细胞凋亡,其机制可能与G_0/G_1期细胞阻滞、抑制DNA合成并激活细胞内caspase-3、caspase-8凋亡途径有关。Objective To study apoptosis effect of matrine （Mat） on acute promyelocytic leukemia cells （NB4） and its mechanism. Methods NB4 cells were treated with Mat of different concentrations （0. 25,0. 5, and 1.0 mg. mL-1 ） for 24,48 ,and 72 hours. The morphologic changes of the cells were observed with Hoechst 33258 staining and proliferation inhibition of NB4 cells was determined with 3- （ 4,5-dimethyl-2-thiazolyl ） -2,5-diphenyl-2-H-tetrazolium bromide （MTT） assay. Cell cycle and apoptosis rate were analyzed by flow cytometry with annexin V-fluorescein isothiocyanate/propidium iodide （FITC/PI） double staining. The relative activity of caspase-3 and caspase-8 were examined with spectrophotometry assay. Results Mat could significantly inhibit the proliferation and induce apoptosis of NB4 cells in a dose- and time-de- pendent manner compared with the normal control （ P 〈 0. 05 ）. Morphology change observation revealed characteristics of cell apoptosis. Mat at the doses of 0. 5 and 1.0 mg.mL-1 could block NB4 cell cycle at G0/G~ stage （57.91% and 83.00 % ） and reduce the proportion of the cells at S stage （40. 95 % and 16. 10% ）, and significantly increase the activity of caspase-3 （77.41±4. 01 and 111.78 ±4. 05）. Further analysis indicated that Mat at the does of 1.0 mg.mL-1 could increase the activity of caspase-8 （44. 98 ±7.63 ） （ P 〈 0. 05 ）. Conclusion Mat could inhibit the proliferation and induce apoptosis of NB4 cells; the mechanism of the effects may be related to the arrest of G0/G1 phase,inhibition of DNA syn- thesis, and activation of caspase-3 and caspase-8 pathway.

关 键 词：苦参碱(Mat) NB4细胞 CASPASE-3 CASPASE-8 细胞凋亡 

分 类 号：R55[医药卫生—血液循环系统疾病]