弓形虫感染对THP-1巨噬细胞极化影响的研究

Effects of the polarization of the macrophage differentiated from THP-1 which infected with Toxoplasmagondii

目的探讨弓形虫感染对体外诱导的人外周血单核细胞(Human acute monocytic leukemia cell line,THP-1)极化特点的影响。方法用佛波酯(PMA)诱导THP-148h使之由悬浮的单核细胞诱导为贴壁的巨噬细胞,体外选择不同时间点感染弓形虫,用Diff染色分析弓形虫在细胞内的增殖情况。Western blot检测极化相关蛋白,Q-PCR检测mRNA的表达情况。结果成功诱导得到贴壁的巨噬细胞模型,感染弓形虫后,M1型巨噬细胞标志性蛋白iNOS及M2型巨噬细胞标志型蛋白Arg-1 36h表达量与对照组差异明显(t=10.23,P<0.05)。Q-PCR的结果显示不同的处理组IL-1、IL-12、iNOS和TNF-α逐渐减少,而IL-10、Arg-1呈逐渐增加,到36h达到顶峰(t=9.587,P<0.05)。结论弓形虫RH株感染THP-1巨噬细胞后可以在胞内生长增殖,THP-1细胞感染后向M2型巨噬细胞方向极化。

To explore the effect of the polarization of the macrophage differentiated from THP-1 which infected with Toxoplasma gondii,monocytes(THP-1)suspended in the culture medium were induced by phorbol ester(PMA)for 48 h to differentiate into adherent macrophages.In vitro experiment at different T.gondii infection time course and the protozoan proliferation rate was studied by using Diff staining.Western blot and Q-PCR were used to detect the expression level of polarization-related proteins and mRNA.The protein expression level of iNOS in M1 macrophage was significantly lower than that in the control group,and the expression of M2 protein Arg-1 was the highest expressed with the highest level at 36 h.Results of Q-PCR showed that the mRNA expression of IL-1,IL-12,iNOS and TNF-αin different treatment groups gradually decreased,but IL-10 and Arg-1 gradually increased and peaked at 36 h(t=9.587,P<0.05).T.gondii can proliferate in the macrophages differentiated from THP-1 cells.After T.gondii infection,it will polarize towards M2 macrophages.