中国优势基因型弓形虫Wh3株诱导人单核巨噬细胞THP-1偏移极化的研究  

Migration of human macrophages THP-1 induced by the dominant genotype of toxoplasma gondii Wh3 strain in China

作　　者：张芳芳 王聪 罗庆礼 沈继龙[1] Zhang Fangfang;Wang Cong;Luo Qingli(Dept of Microbiology and Parasitology,Anhui Provincial Laboratory of Pathogen Biology and Anhui Key Laboratory of Zoonoses,School of Basic Medicine,Anhui Medical University,Hefei 230032;The Affiliated Women and Child Health Care Hospital of Anhui Medical University(Anhui Women and Child Health Care Hospital),Hefei 230001;The Second People’s Hospital of Hefei,Hefei 230011;Dept of Laboratory Center of Pathogenand Immunology,School of Basic Medicine,Anhui Medical University,Hefei 230032)

机构地区：[1]安徽医科大学基础医学院病原生物学教研室,病原生物学安徽省重点实验室,人畜共患病安徽高校省级重点实验室,合肥230032 [2]安徽医科大学附属妇幼保健院(安徽省妇幼保健院),合肥230001 [3]合肥市第二人民医院检验科,合肥230011 [4]安徽医科大学基础医学院病原与免疫学实验中心,合肥230032

出　　处：《安徽医科大学学报》2021年第2期169-175,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家重点基础研究发展计划项目(编号:2010CB530001);国家自然科学基金(编号:81471983)。

摘　　要：目的研究弓形虫Wh3株感染巨噬细胞后,诱导巨噬细胞偏移分化情况。方法获取弓形虫Wh3株、RH株和ME49株,感染人巨噬细胞THP-1,感染后6、24 h分别收集培养上清液和巨噬细胞。ELISA检测培养上清液中巨噬细胞分泌的白细胞介素(IL)-6、IL-10、肿瘤坏死因子(TNF)-α、TGF-β等细胞因子浓度。qRT-PCR分析M1/M2相关基因iNOS、Arginase-1和TNF-α、IL-12、TGF-β、IL-10等细胞因子mRNA转录水平。Western blot分析巨噬细胞iNOS和Arginase-1蛋白表达水平。结果ELISA检测培养上清液中细胞因子含量,发现弓形虫ME49株感染巨噬细胞6、24 h后,IL-6和TNF-α含量最高(P<0.01);弓形虫RH株感染后IL-10和TGF-β含量最高(P<0.01);弓形虫Wh3株感染后培养上清液细胞因子含量介于RH株和ME49株之间(P<0.01)。感染ME49株弓形虫的巨噬细胞24 h后iNOS转录水平及感染6、24 h后TNF-α和IL-12转录水平均升高(P<0.001);RH株感染6、24 h后巨噬细胞的IL-10和Arginase-1的转录水平及感染6 h后TGF-β转录水平升高(P<0.05);同样发现弓形虫Wh3株介于RH株和ME49株之间。Western blot测定iNOS、Arginase-1也得到相似的结果。在感染Wh3株和ME49株弓形虫24 h后,iNOS蛋白表达增高(P<0.0001),而在感染RH株24 h后,Arginase-1表达量升高(P<0.001)。结论弓形虫Wh3株感染后,巨噬细胞表现出介于M1和M2之间的极化方向,且更接近于M2极化方向。Objective To study the polarization of macrophages induced by toxoplasma gondii Wh3 strain. Methods Macrophages THP-1 were infected with toxoplasma gondii Wh3 strain, RH strain and ME49 strain. After 6 hours and 24 hours of infection, the supernatant and macrophages were collected. The concentrations of IL-6, IL-10, TNF-α and TGF-β of the supernatant were detected by ELISA. In macrophages, the mRNA transcription levels of M1/M2 related genes iNOS, arginase-1 and TNF-α, IL-12, TGF-β, IL-10 were detected by qRT-PCR, and the expression levels of iNOS and arginase-1 were analyzed by Western blot. Results The results of ELISA showed that the contents of IL-6 and TNF-α were the highest after infection by ME49 strain 6 hours and 24 hours(P<0.01), and the highest of IL-10 and TGF-β were detected after infected with RH strain(P<0.01). However, the content of cytokines in supernatant infection by Wh3 strain was between RH strain and ME49 strain. Some inflammatory factors and cytokines mRNA in macrophages were detected after 6 hours and 24 hours of infection, and iNOS, TNF-α and IL-12 significantly increased after infected with toxoplasma gondii ME49 strain(P<0.05), arginase-1, IL-10 and TGF-β significantly increased after RH strain infection. It was also found that the levels of mRNA in macrophages infected with Wh3 strain was between RH strain and ME49 strain. After 24 hours of infection, the expression of iNOS protein increased with wh3 and me49 strains of toxoplasma gondii(P<0.000 1), while the expression of arginase-1 increased with RH strain(P<0.001). Conclusion The polarization direction of macrophages was between M1 and M2 after infection with toxoplasma gondii Wh3 strain, and it was closer to M2.

关 键 词：巨噬细胞极化 经典激活途径 旁路激活途径 弓形虫 

分 类 号：R382.5[医药卫生—医学寄生虫学] R392.12[医药卫生—基础医学]