目的探討夏枯草注射液體外抗白血病作用,為臨床應(yīng)用夏枯草注射液治療白血病提供實驗依據(jù).方法采用MTT法測定夏枯草注射液對K562細(xì)胞的增殖抑制率,計算出IC₅₀值;繪制夏枯草注射液(50、100mg/mL)作用于K562細(xì)胞的生長曲線.用倒置顯微鏡、姬姆薩染色法、M TT染色法光鏡下觀察細(xì)胞形態(tài),臺盼藍(lán)拒染法、流式細(xì)胞儀PI染色觀察夏枯草注射液對K562細(xì)胞的促凋亡活性,用免疫細(xì)胞化學(xué)法檢測夏枯草注射液對凋亡相關(guān)基因bcl-2和bax蛋白表達的影響,并用圖文分析系統(tǒng)進行定量分析.結(jié)果夏枯草注射液對K562細(xì)胞增殖具有明顯的抑制作用,且在-定的劑量范圍內(nèi)抑制作用具有明顯的劑量依賴性(r= 0.9850),IC₅₀ 為0.113 mg/mL.夏枯草注射液(50mg/mL)作用于K562細(xì)胞后,倒置顯微鏡、姬姆薩染色、MTT染色光鏡下觀察,均可見典型的凋亡細(xì)胞的形態(tài)學(xué)特征.流式細(xì)胞儀檢測結(jié)果顯示,處理組細(xì)胞凋亡率[(37.15±1.46)%]明顯高于對照組細(xì)胞凋亡率[(5.56±0.68)%](P<0.01).夏枯草注射液(50mg/ mL)作用于K562細(xì)胞48h,bcl-2蛋白表達增強,而bax表達減弱,與對照組相比差異顯著(P<0.01).結(jié)論夏枯草注射液可明顯抑制K 562細(xì)胞增殖,可望成為新的抗白血病藥物,誘導(dǎo)K562細(xì)胞凋亡可能是其發(fā)揮抗腫瘤作用的機制之-.

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.