瘀毒清诱导慢性粒细胞白血病K562细胞株凋亡的研究

Effect of Yudu-qing on the apoptosis of chronic myeloid leukemia K562 cells

目的探讨瘀毒清对诱导慢性粒细胞白血病K562细胞株凋亡的作用。方法采用血清药理学方法，取慢性粒细胞白血病K562细胞株悬液分组，分别加入不含药血清（空白对照组）、含伊马替尼血清、含高剂量瘀毒清血清、中剂量瘀毒清血清、低剂量瘀毒清血清、含伊马替尼加高、中、低剂量瘀毒清血清，不加血清作为正常对照组。分别于干预后第12h、24h、48h和72h通过ArmexinV／PI双染法、流式细胞仪检测不同时间、不同药物组的细胞凋亡率。结果正常对照组的原代细胞有较低凋亡率，而空白对照组K562细胞的凋亡率更低，二者比较差异有统计学意义（P〈0．05）。12h、24h内瘀毒清含药血清低、中、高组凋亡率与正常对照组、空白对照组比较存在统计学差异（P〈O．05）。药物组间相互比较亦有显著差异，并呈一定的时间剂量依赖性。瘀毒清高剂量组72h未见凋亡率（31．48±6．58）进一步升高，与48h比较无统计学差异，与瘀毒清中剂量组72h的诱导凋亡率（27．54±5．89）比较也无统计学差异（P〉0．05）。伊马替尼组12h开始即显示较高的凋亡率（23．80±6．94），与空白对照组细胞的凋亡率相比有统计学差异（P〈0．05）。伊马替尼加瘀毒清各剂量组72h的凋亡率与伊马替尼组、瘀毒清低、中、高组的凋亡率比较均有明显差异（P〈0．05）。结论含瘀毒清血清对体外K562细胞有诱导凋亡作用，其作用呈一定时间、剂量依赖关系：瘀毒清与伊马替尼联合使用有增效作用。

Objective To explore the effect of Yuduqing on the apoptosis of CML K562 cells cultured in vitro and its molecular mechanism. Methods In serologic pharmacological test, the K562 cells were divided into 8 different groups. Serum with imatinib plus Yuduqing （high-dose, middle-dose, low-dose） were added into the cells respectively in the 8 groups of K562 cells. Morphological assessment of apoptosis was performed with optical microscope, the rates of apoptosis and the cell cycles analysis was performed with flow cytometry at 12 h, 24 h, 48 h and 72 h time points respectively after the intervention. Results The primary cell of normal control group had a low rate of apoptosis, while the blank control group K562 cell apoptosis rate was lower, the difference is significant （P〈0.05）. The differences between the rates of apoptosis in high-dose, middle-dose and low-dose Yuduqing groups and those in normal control group and blank control group were significant in 12 hours or 24 hours （P〈0.05）. Drug groups showed significant differences of pair-comparison in groups and a certain time dose dependence. But the rate of apoptosis（31.48±6.58） in k562 cells in high-dose Yuduqing group did not increase further at 72 hours after the intervention and it was not statistically different from that of 48 hours, nor statistically different from that of middle-dose group at 72 hours （27.54±5.89） after the intervention （P〉 0.05）. The rate of apoptosis in k562 ceils in imatinib group （23.80±6.94） was relatively high at 12 hours after the intervention and it was significantly different from that in blank control group （P〈0.05）. The rates of apoptosis in imatinib and Yuduqing （high-dose, middle-dose, and low-dose） groups were significantly higher than those in imatinib group or Yuduqing high-dose, middle-dose, and low-dose）groups （P〈0.05）. Conclusion Serum with Yuduqing could induce apoptosis of K562 cells cultured in vitro and its action was dose-time dependent; Serum with Yuduqing （high-dose and middle-dose） was similar to serum with imatinib in inducing apoptosis ofK562 cells cultured in vitro; Yuduqing could enhance the efficacy ofimatinib.