大黄虫丸对慢性粒细胞白血病K562细胞增殖和凋亡的影响

Effect of Dahuang Zhechong Wan on Chronic Myeloeytic Leukemia K562 Cell Proliferation and Apoptosis

目的从细胞增殖和凋亡角度探讨大黄虫丸治疗慢性粒细胞白血病的可能机理。方法 16只裸鼠随机分为空白组和中药高、中、低剂量组,每组4只。中药高、中、低剂量组分别给予浓度为1.97、0.9、0.45 g/ml的大黄虫丸溶液灌胃,每组0.2 ml/(10 g·d),每日1次,连续7天,空白组给予等量生理盐水。停药次日,分离血清制备大黄虫丸含药血清。应用MTT法检测大黄虫丸含药血清对慢性粒细胞白血病系K562细胞增殖的影响,细胞克隆形成实验检测对细胞克隆形成的影响,用Annexin V FITC/PI法检测对细胞凋亡和细胞周期的影响。结果中药高剂量组在72 h时生长抑制率明显高于中药中、低剂量组(P<0.01)。中药各剂量组克隆形成率较空白组明显下降(P<0.01),中药中、高剂量组克隆形成率明显低于中药低剂量组(P<0.01)。中药高剂量组细胞早期凋亡率、晚期凋亡率及合计凋亡率均较中药低、中剂量组升高明显(P<0.05或P<0.01)。中药低、中、高剂量组G0/G1期细胞显著高于空白组,G2/M期细胞下降(P<0.01),并且以中药高剂量组明显。结论大黄虫丸能抑制K562细胞增殖和诱导其凋亡,将其阻滞于G0/G1期,并且高剂量的大黄虫丸效果最好。

Objective To study, the possible mechanism of Dahuang Zhechong Wan （DHZCW） in treating chronic myeloeytic leukemia （CML） from the angle of cell proliferation and apoptosis. Methods Sixteen nude mice were randomly divided into blank control group and high, medium and low dose traditional Chinese medicine （TCM） groups, with four in each. The high, medium and low dose TCM groups were given DHZCW solution with the concentration of 1.97, 0. 9, 0. 45g/ml by garage, 0. 2ml/ （ 10g ＂d）, once daily for 7 days, while the blank control group received the same volume of normal saline. The medicated serum containing DHZCW was prepared on the next day of withdrawal. The MTY assay, colony formation assay and Annexin V FITC/PI method were applied to detect the impact on the proliferation of CML cell line K562, the formation of colonies and the apoptosis and cell cycle respectively by the medicated serum. Results The growth inhibition rate of the high dose TCM group on 72 h was significantly higher than that of the medium and low dose TCM group （ P 〈 0. 01 ）. The colony forming rate of all TCM groups was significantly lower than the control group （ P 〈 0. 01 ） and that of the high and medium dose TCM group was lower than the low dose TCM group （P 〈0.01 ）. The apoptosls rate in early and late phase, as well as the total apoptosis rate of high dose TCM group were significantly higher than the low dose TCM group （P 〈0. 05 or P 〈 0. 01）. Cells in G0/Gj phase in low, medium and high dose TCM groups were significantly more than that in the blank control group, with less cells in G2/M phase （P 〈0.01 ）, which was most significantly shown in the high dose TCM group. Conclusion DHZCW could inhibit the proliferation of K562 cells and induce the apoptosis by arrest- ing cells in G0/G, phase. Also, best effects achieved with high dose of DHZCW.