夏枯草注射液诱导K562细胞凋亡的实验研究

目的探讨夏枯草注射液体外抗白血病作用,为临床应用夏枯草注射液治疗白血病提供实验依据。方法采用M TT法测定夏枯草注射液对K 562细胞的增殖抑制率,计算出IC50值;绘制夏枯草注射液(50、100 m g/mL)作用于K 562细胞的生长曲线。用倒置显微镜、姬姆萨染色法、M TT染色法光镜下观察细胞形态,台盼蓝拒染法、流式细胞仪P I染色观察夏枯草注射液对K 562细胞的促凋亡活性,用免疫细胞化学法检测夏枯草注射液对凋亡相关基因bcl-2和bax蛋白表达的影响,并用图文分析系统进行定量分析。结果夏枯草注射液对K 562细胞增殖具有明显的抑制作用,且在一定的剂量范围内抑制作用具有明显的剂量依赖性(r=0.985 0),IC50为0.113 m g/mL。夏枯草注射液(50 m g/mL)作用于K 562细胞后,倒置显微镜、姬姆萨染色、M TT染色光镜下观察,均可见典型的凋亡细胞的形态学特征。流式细胞仪检测结果显示,处理组细胞凋亡率(37.15±1.46)%]明显高于对照组细胞凋亡率(5.56±0.68)%](P<0.01)。夏枯草注射液(50 m g/mL)作用于K 562细胞48 h,bcl-2蛋白表达增强,而bax表达减弱,与对照组相比差异显著(P<0.01)。结论夏枯草注射液可明显抑制K 562细胞增殖,可望成为新的抗白血病药物,诱导K 562细胞凋亡可能是其发挥抗肿瘤作用的机制之一。

Inductive effect of Prunella vulgaris Injection on K562 cells apoptosis

Objective To investigate the antileukemia effect ofPrunella vulgarisInjection(PVI)in order to offer experimental data for the treatment of leukemias with PVI in clinic.Methods Inhibition of PVI on K562 cell proliferation and IC₅₀ value was assayed by MTT assay.The growth curve of K562 cells was drawn also.The cellular morphology was observed by invert micro-scope,Giemsa staining,and MTT assay.The apoptosis induced the effect of PVI to K562 cells was observed by trypan-blue exclusion test and flow cytometer(FCM).The expression of apoptosis related protein bcl-2 and bax was measured by immunocytochemistry,and the quan-titative analysis was made with figure analysis system.Results PVI obviously inhibited the proliferation of K562 cells in a dose-dependent manner(r-0.9850).The IC₅₀ was 0.113 mg/mL.After K562 cells were treated with PVI(50mg/mL),the morphologi-cal characters of apoptosis were observed.The results of FCM showed that the apoptosis rats(37.15±1.46)%]in the treatment group were significant higher than that in control group I-(5.56-t-0.68)%](P<0.01).After K562 cells were treated by PVI(50 mg/mL)for 48 h,the expression of protein bcl-2 was up-regulated,and the expression of protein bax was down-regulated.The differences between the treatment group and control group are significant(P<0.01).Conclusion PVI can inhibit the proliferation of K562 cells and may be a new antileukemia drug.PVI can induce the apoptosis of K562 cells,which may be one of antileukemia mechanisms of PVI.