弓形虫速殖子感染对HeLa细胞凋亡的影响

Effect of Toxoplasma gondii tachyzoite infection on HeLa cell apoptosis

目的:探讨不同数量弓形虫速殖子感染对HeLa细胞凋亡的影响。方法:24孔板培养HeLa细胞16 h,各孔接种不同数量弓形虫速殖子继续培养4 h,在培养液中加入终质量浓度为0.5μg/mL放线菌素D诱导凋亡,于加放线菌素D后8、12、24、36 h收集细胞,采用Hoechst 33258染色法和Annexin V-FITC/PI细胞凋亡检测试剂盒检测细胞凋亡率。结果:当虫体量/细胞量(MOI)≤1时,弓形虫感染表现为抑制放线菌素D诱导的HeLa细胞凋亡;随着虫体数量增加(MOI>1)和感染时间延长(≥24 h),虫体感染则表现为促进HeLa细胞凋亡。结论:弓形虫感染可影响体外培养的HeLa细胞的凋亡,影响程度与感染虫体数量相关。

Objective:To explore HeLa cell apoptosis when infected with different number of T.gondii tachyzoites.Methods:HeLa cells were cultured in 24-wells plates for 16 h and incubated with different number of T.gondii tachyzoites.Mter infected for 4 h,the concentration of 0.5 μg/mL actinomycin D was added into the medium.The cells were collected in 8,12,24 and 36 h.The apoptosis of HeLa cell was determined by Hoechst 33258 stain and Annexin V-FITC/PI method.Results:When the rate of parasites/ cells (multiplicity of infection,MOI) was no more than 1 (MOI ≤ 1),the parasites could suppress the cell apoptosis.When MOI ＞ 1 and the infection time over 24 h,the parasites could accelerate the cells apoptosis.Conclusion:T.gondii tachyzoite infection could affect the HeLa cell appotosis in vitro,and the incidence was related to the number of tachyzoites.