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  

The effects of artesunate on Eimeria tenella sporozoites invading DF-1 cells

作　　者：焦金英[1,2] 赵昕 承南[1,2] 刘明江 李金贵 夏前贤[1,2] 

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]江苏省动物重要疫病与人畜共患病防控协同创新中心,江苏扬州225009

出　　处：《中国预防兽医学报》2018年第2期147-151,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金资助项目(31272548);江苏省高校优势学科建设工程项目

摘　　要：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(2.5μmol/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SP 2 h,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2 h和12 h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。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 ~mol/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 exlgerimental basis for research on anti coccidiosis mechanism of artemisinin drugs.

关 键 词：青蒿琥酯 柔嫩艾美尔球虫 子孢子 侵入率 

分 类 号：S852.7[农业科学—基础兽医学]