双氢青蒿素与雷公藤甲素对神经胶质瘤细胞的抑制作用

Inhibiting effects of dihydroartemisinin and triptolide on glioma cells

目的探讨两种天然药物双氢青蒿素(DHA)与雷公藤甲素(TP)单独及联合用药对神经胶质瘤细胞体外抑制和体内治疗的作用。方法 CCK-8法检测DHA与TP单独及联合用药对神经胶质瘤细胞活性的抑制作用。Hochest 33258染色观察给药后细胞核的形态变化,采用Rhodamine123和PI染色技术在流式细胞仪下分别检测细胞线粒体膜电位和细胞周期的情况。建立神经胶质瘤小鼠移植瘤模型,给药后分别监测小鼠体质量以及肿瘤体积大小并作统计学分析。苏木精-伊红(H-E)染色主要脏器的病理切片,观察药物的在体毒性。结果 12μg/m L的DHA与TP单独及联合用药处理神经胶质瘤C6和U87细胞48 h可明显抑制细胞的活性,抑制率分别达到85%和80%。DHA与TP单独及联合用药组分别都引起细胞核固缩,细胞线粒体膜电位下降明显,联合用药组的细胞线粒体膜电位的下降明显高于单独用药组(P<0.01)。DHA与TP联合处理细胞主要使细胞周期阻滞在Sub-G1和G2/M期。在体实验结果显示:相比于对照组和单独给药组,联合用药组更能够有效抑制肿瘤的生长(P<0.01)。病理切片H-E染色发现联合用药几乎没有在体毒性。结论DHA与TP联合用药对神经胶质瘤细胞具有显著的协同抑制效果,并且在体治疗效果也非常显著,为临床使用天然药物治疗神经胶质瘤提供一个新的方法和思路。

Objective To investigate the effects of alone and combined administration of two natural drugs dihydroartemisinin（DHA） and triptolide（TP） on in vitro inhibition and in vivo treatment of glioma.Methods C C K-8 method was adopted to detect the inhibitory effect of alone and combined administration of DHA and TP on the viability of glioma cells.Hochest 33258 staining was used to observe changes of cell nucleus after administration.Rhodamine123 and PI staining was adopted to detect the mitochondrial membrane potential and cell cycle.The transplanted tumor model of mice with glioma was established.The body weight and size of tumor were observed after administration and were statistically analyzed.Pathological sections of main organs were H-E stained and the in vivo toxicity of drugs was observed.Results Alone and combined administration of DHA and TP of 12 μg/mL for treating glioma cells C6 and U87 for 48 h significantly inhibited the cell viability and inhibition rates were 85%and 80%,respectively.Alone and combined administration of DHA and TP could cause the karyopyknosis and significant decline of mitochondrial membrane potential.The decline of mitochondrial membrane potential caused by combined administration was more than that caused by alone administration（P〈0.01）.Combined administration of DHA and TP mainly arrested the cell cycle at Sub-G_1and G_2/M phases.The results of in vivo tests suggested that combined administration could inhibit the growth of tumor more effectively than alone administration and controls（P 0.01）.H-E stained pathological sections showed that combined administration had little in vivo toxicity.Conclusion Combined administration of DHA and TP has significant synergetic inhibition effect on glioma cells and the in vivo therapeutic effect is also remarkable,which provides a new method for clinical treatment of glioma with natural drugs.