体外弓形虫感染小鼠巨噬细胞对其极化的影响  

The effect of Toxoplasma gondii infection on mouse macrophage polarization in vitro

作　　者：祝瑶[1] 胡守萍[1] 张交儿[1] 张卓[1] 郑君[1] 贾洪林[1] 何希君[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2016年第6期475-478,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：兽医生物技术国家重点实验室(SKLVBP201430)

摘　　要：为探索体外弓形虫感染小鼠巨噬细胞对其极化的影响,本研究采用M-CSF诱导分离的小鼠骨髓细胞分化为巨噬细胞,在体外以弓形虫感染骨髓来源的巨噬细胞,分别于感染后0 h、6 h、12 h、24 h、36 h、48 h收集细胞和细胞培养液,利用Griess法和Arginase活性试验对i NOS和Arginase-1的酶活性进行检测。通过荧光定量PCR和western blot方法分别从m RNA水平和蛋白水平测定巨噬细胞中i NOS和Arginase-1的表达量。结果显示,弓形虫感染巨噬细胞后细胞中的Arginase-1的m RNA和蛋白含量均显著上调表达,i NOS的m RNA虽有短暂小幅度上调,但蛋白的含量无明显变化。本研究表明弓形虫感染巨噬细胞后能够明显诱导静息状态的巨噬细胞朝向M2型的方向极化,从而改变巨噬细胞的表型和功能。本研究为进一步阐明弓形虫与机体免疫系统相互作用提供了实验依据。This study was designed to explore the effects of Toxoplasma gondii infection on mouse macrophage polarization/n vitro. The isolated bone marrow cells from mice were induced with M-CSF to differentiate into macrophages, and the bone marrow-derived macrophages were infected with T.gondii, while cells and cell culture medium were collected at 0, 6, 12, 24, 36 and 48 hours post inoculation. The activity of iNOS and Arginase-1 was detected by arginase activity assay and Griess assay. In addition, the expression of iNOS and Arginase-1 was further detected by real-time PCR and western blot in the mRNA and protein levels, which showed the expression of mRNA and protein levels of Arginase-1 was significantly increased in macrophages infected with T.gondii. Althought there was an increase in the iNOS mRNA level at 6 hours after infection, the expression iNOS protein level of iNOS was not significantly different to the negative control at all the time points. Hence, when resting state bone marrow-derived macrophages were infected with T.gondii, they were differentiated into type 2, but not type 1 polarization. At the same time, the phenotype and function of macrophages were altered afterward. The study contributes to further elucidate the interacting between T.gondii and host＇ immune system.

关 键 词：弓形虫 巨噬细胞 极化 

分 类 号：S852.7[农业科学—基础兽医学]