调控宿主细胞自噬对弓形虫在宿主细胞内增殖的影响

Influence of regulation of host cell autophagy on the proliferation of Toxoplasma gondii in host cells

目的研究细胞自噬在弓形虫增殖复制中的作用。方法分别用自噬抑制剂Bafilomycin A1及自噬诱导剂氯化锂作用于人胚胎成纤维细胞(HEF),然后将弓形虫(RH株)速殖子悬液分别以虫/细胞2∶1、4∶1、8∶1、16∶1感染HEF细胞,作用1、2、4、8、24、48、96 h。采用吖啶橙(AO)荧光染色和丹酰尸胺(MDC)荧光染色检测HEF细胞自噬情况,采用Giemsa染色检测不同时间点的弓形虫感染复制动力学。结果 AO和MDC荧光染色结果表明自噬抑制剂Bafilomycin A1及自噬诱导剂氯化锂可分别抑制和促进HEF细胞自噬,Giemsa染色结果显示使用自噬诱导剂氯化锂可以促进弓形虫在HEF细胞内的增殖,使用自噬抑制剂Bafilomycin A1可以抑制弓形虫在HEF细胞内的增殖。结论调控宿主细胞自噬可调控弓形虫增殖复制。

Objective To study the role of autophagy in the replication process of Toxoplasma proliferation. Meth-ods As the experimental groups, these cells were infected by Toxoplasma gondii tachyzoites at given MOI （2 ∶ 1, 4 ∶ 1 ,8 ∶ 1 ,16 ∶ 1 ） . Host cell autophagy was detected through acridine orange staining and MDC fluorescence stai-ning at different time points （1, 2, 4, 8, 24, 48, 96 hs post infection）. Detect the condition of HEF cells autoph-agy with acridine orange fluorescence staining and MDC fluorescence staining, and detect the replication kinetics of Toxoplasma gondii infection at different time points using Giemsa staining. Results The results of acridine orange and MDC fluorescence staining showed that autophagy inhibitors and inducers could inhibit and promote the autoph-agy of HEF cells respectively. From the results of Giemsa staining, it was found that the proliferation of Toxoplasma gondii in HEF cells could be promoted with autophagy inducers and be inhibited with autophagy inhibitors. Conclu-sion The regulation on autophagy of host cell could regulate the proliferation and replication of Toxoplasma gondii.