纳米甲硝唑体外抗滴虫效果及机制的初步研究

Preliminary study on trichomonacidal activity and mechanism of metronidazole-loaded nanoparticles in vitro

目的初步研究纳米甲硝唑体外抗阴道毛滴虫的效果及药物作用机制。方法以肝浸汤培养基培养阴道毛滴虫,通过四甲基偶氮唑盐微量酶反应比色法(MTT法)及传统的记数法观察纳米甲硝唑的体外杀虫效果,利用4,6-联脒-2-苯基吲哚(DAPI)荧光染色法观察纳米药物作用后滴虫细胞核的变化。设甲硝唑原料药组、纳米甲硝唑组、纳米载体组和不加药物处理的对照组。结果纳米甲硝唑及甲硝唑原料药均有抑杀滴虫的作用,与不加药物处理的对照组及纳米载体组比较,差异均有显著性(P均<0.05),杀虫效果随着药物浓度的增加及作用时间的延长而增强。且在甲硝唑浓度相同的情况下,纳米甲硝唑抗滴虫的效果优于甲硝唑原料药,该组滴虫的相对抑制率及死亡率均高于甲硝唑原料药组(P<0.05)。荧光显微镜下观察,使用纳米甲硝唑后,滴虫细胞核出现核固缩、核碎裂等类似凋亡细胞的一系列改变。结论纳米甲硝唑具有较强的体外抗滴虫效果,杀虫效果优于普通甲硝唑,且很可能通过诱导或加速滴虫细胞发生凋亡样死亡而发挥药物作用。

Objective To study the trichomonacidal activity of metronidazole-loaded nanoparticles against Trichomonas vaginalis in vitro. Methods MTT colorimetric assay and classical microscopic counting method were used to determine the inhibitory effect of metronidazole-loaded nanoparticles. The morphological nuclear changes were observed by DAPI staining. Inhibitory effect was observed from metronidazole standard group, metronidazole-loaded nanoparticles group,drug-carrier group, and the control. Results There was significant difference between the metronidazole-loaded nanoparticles group and the control group （P〈0.05）. The group of metronidazoleloaded nanoparticles also had a significant difference with the drug-carrier group （P〈0.05）. The anti-trichomonad activity increased with the drug concentration and time. Additionally, with metronidazole equivalent in concentration to standard solution of metronidazole, drug-loaded nanoparticles had a better anti-trichomonad activity than metronidazole standard （P〈0. 05）. After metronidazole-loaded nanoparticles treated, fragmentation of the cell nuclei was observed by the stain with the specific fluorescent dye DAPI. Conclusion Metronidazole-loaded nanoparticles may induce the apoptosis-like death of T. vaginalis＇ cell.