参附注射液、高乌甲素注射液诱导白血病细胞株HL-60分化、凋亡的研究

A Study on Shenfu Injection and Lappaconitine Hydrobromide Injection Inducing HL-60 Differentiation and Apoptosis

【目的】观察参附注射液及高乌甲素注射液诱导白血病细胞株HL-60分化、凋亡的作用,初步验证温阳法治疗肿瘤的理论。【方法】采用人类急性白血病细胞株HL-60为模型,设参附注射液组(终浓度分别为12.5、25、50、100μL/mL),高乌甲素组(终浓度分别为25、50、100μL/mL),亚砷酸钠对照组(终浓度分别为0.25、2、15μmol/L)及阴性对照组;检测细胞增殖、细胞形态、硝基四氮唑蓝(NBT)还原能力、细胞周期分布、凋亡率等指标。【结果】参附注射液作用后HL-60细胞增殖受抑制,高乌甲素注射液对HL-60的生长无抑制作用;经药物处理60 h后HL-60细胞均出现程度不同以凋亡为主的形态改变;低浓度参附注射液(12.5μL/mL)、高乌甲素注射液(50μL/mL)作用60 h后HL-60细胞NBT还原能力增强,而其他浓度组NBT还原能力反未增强;药物处理60 h后,100μL/mL高乌甲素液及25.50μL/mL参附液流式细胞仪检测直方图上呈现亚二倍体凋亡峰,凋亡率呈不同程度增高。【结论】参附注射液、高乌甲素注射液对人类急性白血病细胞株HL-60具有不同程度的诱导分化和凋亡的双重作用,为温阳法治疗肿瘤提供了一定的理论依据。

Objective To observe the effect of Shenfu Injection（SI） and lappaconitine hydrobromide injection（LHI） on inducing HL-60 differentiation and apoptosis,and to supply the experimental evidence for warming-yang method in treating tumor.Methods Human acute myelocytic leukemia cell line of HL-60 was established.SI groups（at the final concentrations of 12.5,25,50 and 100 μL/mL respectively）,LHI groups（at the final concentrations of 25,50 and 100 μL/mL respectively）,sodium arsenite control groups（at the final concentrations of 0.25,2 and 15 μmol/mL respectively）,and negative control group were set up.The parameters of cell proliferation,cell morphology,nitroblue-tetrazolium（NBT） reduction,cell cycle and apoptotic rate were observed.Results SI inhibited HL-60 proliferation,but LHI had no inhibition on HL-60 growth after co-culture.Apoptosis of HL-60 in different degrees was found in SI groups and LHI groups after co-culture for 60 hours.NBT reduction ability was increased in low-dose SI group and low-dose LHI group after treatment for 60 hours,but NBT reduction ability remained unchanged in high-dose LHI group and high-dose SI group.The sub-G1 peak was found in flow-cytometry test and the apoptosis rate was increased to different degrees.Conclusion SI and LHI have biphasic effect on inducing differentiation and apoptosis of HL-60.And this will supply the experimental evidence for warming-yang method in treating leukemia.