青蒿琥酯对鸡柔嫩艾美尔球虫子孢子侵入DF-1细胞的影响

摘要

In order to study the effect of artesunate (ARS) on E.tenella sporozoites invading cells,DF-1 cells and sporozoites were treated respectively by ARS.After porozoites labeled by special fluorescent dye,the apoptotic rate of DF-1 cells,mitochondrial membrane potential of sporozoites and invasion rate of sporozoites were detected by flow cytometry.The results showed that there was no significant difference in apoptotic rate of DF-1 cells between treated group by ARS (2.5 μmol/L) for 12 hrs and untreated treatment.Comparing to the control,ARS directly treated sporozoites for 2hrs,the mitochondrial membrane potential decreased by 33.97±8.40％ and the invasion rate of the sporozoites declined by 4.13±3.97％.The cell were treated with ARS for 2hrs or 12hrs before adding the sporozoites,the invasion rate of the sporozoites accordingly decreased by 7.23±2.74％ and 53.67±0.74％,respectively.The cells were mixed with sporozoites and then treated by ARS for 12hrs,the invasion rate of the sporozoites declined by 56.30± 1.81％.The cells and sporozoites were simultaneously treated by ARS and bongkerkic acid,the invasion rate enhanced by 9.70±0.60％ (p＜0.01).Therefore,the results indicated that ARS obviously inhibited the invasion rate of sporozoites mainly via acting with the sites on cellular membrane,and this action had a significant time-effect.This study provides experimental basis for research on anti coccidiosis mechanism of artemisinin drugs.%为研究青蒿琥酯(ARS)对柔嫩艾美耳球虫子孢子(SP)侵入DF-1细胞的影响,本实验利用ARS分别处理SP和DF-1细胞,前者经特定荧光标记后,流式细胞仪检测细胞凋亡、SP线粒体膜电位和侵入率.结果显示,ARS (2.5 μmol/L)处理DF-1细胞12 h与未处理组相比,细胞凋亡率无明显差异.与空白对照组相比,ARS直接处理SP2h,可使SP线粒体膜电位下降33.97±8.40％,并使其对细胞的侵入率下降4.13±3.97％.ARS直接处理细胞2h和12h后,再加入SP,则其侵入率分别下降7.23±2.74％和53.67±0.74％.细胞和SP混合后用ARS处理12 h,其侵入率下降56.30±1.81％.若在ARS处理的同时加入凋亡抑制剂米酵菌酸,可以使SP的侵入率提高9.70±0.60％(p＜0.01).这表明ARS作用于细胞后对SP侵入的抑制作用更为明显,并具有显著的时间效应.本研究为青蒿素类药物抗球虫作用机理研究提供实验依据.