摘要
目的 用SYBR green Ⅰ法和流式细胞术评价3种青蒿素类药物对体外培养恶性疟原虫的抗疟活性. 方法 采用SYBR green Ⅰ法观察不同浓度的青蒿素（arteminisinin,ART）、蒿甲醚（artemether,AM）及双氢青蒿素（dihydroartemisinin,DHA）对体外培养的恶性疟原虫增殖的抑制作用.ART、AM、DHA的抑制浓度依次为6、6×10^-1、6×10^-2、6×10^-3、6×10^-4 μmol/L,4、4×10^-1、4×10^-2、4×10^-3、4× 10^-4 μmol/L及3×10^-1、3×10^-2、3×10^-3、3×10^-4、3×10^-5 μmol/L,设空白组与对照组.体外药物作用72 h后,SYBR green Ⅰ法测得其抑制率,用50％抑制浓度（50％ inhibitory concentraton,IC50）及95％抑制浓度（95％ inhibitory concentraton,IC95）反映其抑制作用,并与流式细胞术所测得的疟原虫DNA含量的变化进行比较. 结果 SYBR green Ⅰ法结果显示其抑制率随药物浓度的增加而增大,ART、AM与DHA的IC50值分别为11.29、3.32、0.66 nmol/L,IC95值分别为42.55、17.89、4.02 nmol/L;流式细胞术结果中其DNA含量随药物浓度的降低而增加,与SYBR green Ⅰ法结果一致且更具直观性. 结论 SYBR green Ⅰ法的灵敏性与流式细胞术的直观性可以满足体外测定抗疟药对疟原虫抑制作用的需要,均可适于抗疟药物体外活性评价;ART、AM与DHA的体外抗疟活性由高到低的次序为DHA＞AM＞ART.
Objective To evaluate the antimalarial activity against Plasmodiumfalciparum in vitro by SYBR green Ⅰ and flow cytometry.Methods SYBR green Ⅰ was used to evaluate the inhibition to proliferation of Plasmodium falciparum in vitro treated by artimisinin（ART）,artemether（AM） and dihydroartemisinine（DHA） at different concentrations.The intervention concentrations of ART,AM and DHA were 6,6×10^-1,6×10^-2,6×103,6×10^-4μmol/L,4,4×10^-1,4×10^-2,4×10^-3,4×10^-4 μmol/L,and 3×10^-1,3×10^-2,3×10^-3,3×10^-4,3×10^-5 μmol/L respectively,a blank control group was set up.After 72 h of drug intervention in vitro,the inhibition rate was measured using SYBR green Ⅰ,the 50% inhibitory concentration （IC50） and 95% inhibition concentration （IC95） were used to reflect the inhibition activity,then compared with the changes of DNA content of malaria parasites measured by flow cytometry.Results SYBR green Ⅰ showed the inhibition rates increased as dose of ART increased,The IC5o values of ART,AM and DHA were 11.29,3.32,0.66 nmol/L; IC95 values were 42.55,17.89,4.02 nmol/L respectively.At the same time,the results of the flow cytometry showed the content of DNA decreased with the increase of drug concentration,agreeing with the results of SYBR green Ⅰ and being objectively perceived.Conclusion The sensitivity of SYBR green Ⅰ and intuitiveness of flow cytometry could meet the requirements of evaluation on antimalarial activity of artemisinin-based antimalarial drugs against Plasmodiumfalciparum in vitro; The order from high to low of antimalarial activity is DHA〉AM〉ART.
出处
《国际医学寄生虫病杂志》
CAS
2014年第4期218-222,共5页
International JOurnal of Medical Parasitic Diseases
基金
国家自然科学基金（81373364）
