青蒿琥酯联合三氧化二砷对NB4细胞增殖与凋亡的影响  

Effect of combination of artesunate and arsenic trioxide on the proliferation and apoptosis of NB4 cells

作　　者：尹婷 任汝静[1] 李晓强 赵丽凤 程金来 沈建英 谭余庆[1] Yin Ting;Ren Rujing;Li Xiaoqiang;Zhao Lifeng;Cheng Jinlai;Shen Jianying;Tan Yuqing(Master of 2016,Safety Evaluation Center of Chinese Materia Medica,Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100029,China)

机构地区：[1]中国中医科学院中药研究所中药安全评价中心,北京100029

出　　处：《国际中医中药杂志》2018年第11期1049-1054,共6页International Journal of Traditional Chinese Medicine

基　　金：中央级公益性科研院所基本科研业务费专项资金资助（ZXKT15026）

摘　　要：目的 研究青蒿琥酯联合三氧化二砷对NB4细胞增殖与凋亡的影响。方法 将不同浓度的青蒿琥酯和三氧化二砷分别作用于NB4细胞48 h，应用MTT法检测细胞增殖情况。将细胞分为空白对照组、青蒿琥酯组、三氧化二砷组、三氧化二砷联合青蒿琥酯组。青蒿琥酯组以0.4 μmol/L青蒿琥酯进行干预，三氧化二砷组以1 μmol/L三氧化二砷干预，三氧化二砷联合青蒿琥酯组以0.4 μmol/L青蒿琥酯及1 μmol/L三氧化二砷进行干预。采用流式细胞术检测细胞周期和细胞凋亡，采用蛋白免疫印迹法检测细胞Bcl-2与Bax蛋白表达。结果 MTT检测显示，与对照组比较，青蒿琥酯0.25、0.50、1.00、2.00、4.00 μmol/L组细胞增殖抑制率[分别为（19.26±3.59）%、（36.53±2.67）%、（61.32±2.50）%、（70.30±3.15）%、（86.92±0.02）%]升高（P＜0.05）；三氧化二砷1、2、4、8、16 μmol/L组细胞增殖抑制率[分别为（12.69±2.43）%、（64.26±2.02）%、（85.10±2.67）%、（92.06±2.21）%、（93.67±3.36）%]升高（P＜0.05）；与三氧化二砷组比较，三氧化二砷联合青蒿琥酯组增殖抑制率[（40.17±5.49）%比（32.23±3.52）%]、细胞G0/G1期细胞比例[（74.20±1.43）%比（66.14±1.78）%]、细胞凋亡率[（58.00±2.41）%比（34.57±1.22）%]升高（P＜0.05）；细胞Bcl-2蛋白[（0.45±0.09）比（1.03±0.10）]表达降低（P＜0.05），Bax蛋白[（1.35±0.09）比（1.13±0.09）]表达升高（P＜0.05）。结论 青蒿琥酯可明显增强三氧化二砷对NB4细胞的增殖抑制和诱导凋亡作用，其机制可能与下调抗凋亡Bcl-2蛋白表达及上调促凋亡蛋白Bax表达有关。Objective To investigate the effects of artesunate combined with arsenic trioxide （ATO） on the proliferation and apoptosis of NB4 cells. Methods The NB4 cells were treated with different concentrations of artesunate and arsenic trioxide respectively for 48 h. The cell proliferation was detected by methyl thiazolyl tetrazolium （MTT） method. The cells were divided into 4 groups： control group, artesunate group, arsenic trioxide group, and the combination of artesunate and arsenic trioxide group. The cell cycle and apoptosis were detected by flow cytometry （FCM）. The protein expression levels of Bcl-2 and Bax were detected by Western blot. Results The MTT results showed that compared with the control group, the proliferation inhibition rates of 0.25, 0.50, 1.00, 2.00, 4.00 μmol/L artesunate group （19.26% ± 3.59%, 36.53% ± 2.67%, 61.32% ± 2.50%, 70.30% ± 3.15%, 86.92 ± 0.02%） significantly increased （P〈0.05）; the proliferation inhibition rates of 1, 2, 4, 8, 16 μmol/L arsenic trioxide group （12.69% ± 2.43%, 64.26% ± 2.02%, 85.10% ± 2.67%, 92.06% ± 2.21%, 93.67% ± 3.36%） significantly increased （P〈0.05）; and the proliferation inhibition rate （40.17% ± 5.49% vs. 32.23% ± 3.52%） of combination of artesunate and arsenic trioxide group significantly higher than the arsenic trioxide group （P〈0.05）. Compared with the arsenic trioxide group, the percentage of G0/G1 phase cells （74.20% ± 1.43% vs. 66.14% ± 1.78%）, the apoptosis rate （58.00% ± 2.41% vs. 34.57% ± 1.22%）, and the expression level of Bax protein （1.35 ± 0.09 vs. 1.13 ± 0.09） in the combination of artesunate and arsenic trioxide group significantly increased （P〈0.05）, the expression level of Bcl-2 protein （0.45 ± 0.09 vs. 1.03 ± 0.10） in the combination of artesunate and arsenic trioxide group significantly decreased （P〈0.05）. Conclusions Artesunate can significantly enhance the proliferation inhibition and apoptosis induced by arsenic trioxide on NB4 cells. The possible

关 键 词：细胞增殖 细胞凋亡 青蒿琥酯 三氧化二砷 基因 BCL-2 bcl-2-相关x蛋白 NB4 细胞 

分 类 号：R285[医药卫生—中药学]