六神丸诱导HL-60细胞凋亡的实验研究

An Experimental Study on the Apoptosis of HL - 60 Cells Induced by Liushen Wan

目的:从细胞凋亡角度探讨六神丸治疗白血病的机理,为临床应用提供依据。方法:分别制备六神丸乙醇提取剂和六神丸含药血清,用MTT法检测六神丸对HL-60细胞增殖的抑制作用,吉姆萨染色观察细胞凋亡的形态学变化,原位末端标记(POD)法检测细胞凋亡现象。结果:六神丸乙醇提取剂对HL-60细胞的增殖有明显的抑制作用,且呈现浓度依赖关系;中剂量组所得六神丸含药血清对HL-60细胞增殖的抑制作用优于低剂量组和高剂量组。吉姆萨染色显示六神丸乙醇提取剂和六神丸含药血清处理过的HL-60细胞均出现典型的凋亡形态。原位末端标记表明,六神丸乙醇提取剂诱导HL-60细胞凋亡在一定范围内呈现时间-浓度依赖关系;六神丸含药血清诱导HL-60细胞凋亡作用优于As_2O_3血清组和空白血清组(P<0.01)。结论:六神丸体外诱导HL-60细胞凋亡可能是六神丸治疗白血病的作用机制之一。

Objective: From the viewpoint of apoptosis to explore the mechanism of Liushen Wan(LW) for leukemia, so as to provide the evidence for clinical application of LW. Methods: Alcoholic LW extract and serum containing LW were prepared individually. The inhibition of HL - 60 cellular proliferation by LW was datermined by MTT method; the morphological change of apoptotic cell was ob- served by Giemsa stain; and the apoptotic phenomenon was detected by orthotopic terminal label. Results: The alcoholic LW extract can markedly inhibit the proliferation of HL - 60 cells in a dose - de- pendent manner. The action of inhibiting HL - 60 cell proliferation by moderate dosage of serum containing LW is superior than that of low dosage and high dosage. Giemsa stain showed that typical apop- totic structure appeared in HL - 60 pretreated by alcoholic LW ex- tract and serum containing LW. The orthotopic terminal label showed that the alcoholic LW extract induced apoptosis of HL - 60 cells in a time - dose dependent manner. The action of inducing HL - 60 cells apoptosis by serum containing LW is stronger than that of As_2O_3 - serum group and hlank serum group(P < 0.01). Conclusion: LW inducing HL - 60 cells apoptosis in vitro may be one of the mechanisms for the treatment of leukemia.