青蒿琥酯对小鼠骨髓造血干祖细胞分化的抑制与毒性作用

目的 观察青蒿琥酯(artesunate, AS) 对小鼠骨髓造血干/祖细胞分化的影响。方法 取小鼠骨髓干细胞进行体外液体或半固体定向培养并加入不同浓度的AS, 光镜下计数半固体培养的红系集落形成单位(CFU-E) 与红系爆式集落形成单位(BFU-E) 数量, 流式细胞术检测液体定向培养的细胞凋亡和线粒体膜电位变化, 同时进行DNAladder 凝胶电泳实验。结果 AS 可显著抑制CFUE与BFU-E 集落的生成(P <0.05) 并具有一定的量效关系;DNA ladder 凝胶电泳结果显示0.01 ~1.30 mmol/L 的AS 可明显诱导细胞凋亡, 流式细胞仪检测细胞晚期凋亡 死亡率具有统计学意义(P <0.05);线粒体膜电位的下降程度也与AS 浓度正相关(r=0.546, P=0.018)。结论 AS 对小鼠骨髓造血干/祖细胞的增殖和分化具有明显抑制作用, 小剂量AS 可诱导小鼠骨髓造血干/祖细胞发生凋亡, 大剂量可直接引起细胞坏死。

Inhibitive and toxic effects of artesunate on murine bone marrow hematopoietic stem and progenitor cells

AIM: To observe the influences of artesunate (AS) on murine bone marrow hematopoietic stem and progenitor cells.METHODS: Murine bone marrow hematopoietic stem and progenitor cells were cultured in vitro with different concentration of AS. The numbers of erythrocyte colony-forming unit(CFUE) or burst-forming unit (BFU-E) in liquid or semisolid culture were counted through light microscope.Apoptosis of cultured cells and the changes of mitochondrial membrane potential were detected by flow cytometry (FCM) analysis, meanwhile, DNA ladder gel electrophoresis were conducted.RESULTS: AS could significantly inhibit the formation of CFU-E and BFU-E (P <0.05), and the inhibition effect showed a dose-dependent manner.The result showed that AS could significantly induce cell apoptosis at the concentration ranges from 0.01 mmol/L to 1.30 mmol/L by DNA fragmentation gel electrophoresis assay, and the advanced stage death rate by FCM analysis had statistical significance (P <0.05).There was a positive correlation between AS dose and the decrease of mitochondria membrane potential (r=0.546, P=0.018).CONCLUSION: AS can significantly inhibit the proliferation and differentiation of murine bone marrow hematopoietic stem and progenitor cells.AS can induce apoptosis at low dose and cause cell necrosis at high dose.