刚地弓形虫钙依赖蛋白激酶9对小鼠Ana-1巨噬细胞功能的影响  

Effects of Calcium Dependent Protein Kinase 9 of Toxoplasma gondii on the Functions of Mouse Ana-1 Macrophages in vitro

作　　者：马群山[1] 刘欣超[1] 孙晓妮[1] 王帅[1] 徐立新[1] 宋小凯[1] 严若峰[1] 李祥瑞[1] 

机构地区：[1]南京农业大学动物医学院,南京210095

出　　处：《畜牧兽医学报》2017年第4期731-739,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：农业部公益性行业专项(200903036)

摘　　要：本试验旨在探究刚地弓形虫钙依赖蛋白激酶9(CDPK9)对小鼠Ana-1巨噬细胞活性及功能的影响。利用激光共聚焦技术验证CDPK9与Ana-1巨噬细胞的结合情况;采用细胞计数试剂盒(cell counting kit-8,CCK-8)检测不同浓度CDPK9处理24h对Ana-1巨噬细胞增殖活性的影响;利用流式细胞术检测不同浓度CDPK9处理48h对Ana-1巨噬细胞凋亡及吞噬功能的影响;采用总一氧化氮试剂盒检测不同浓度CDPK9处理48h对Ana-1巨噬细胞NO分泌量的影响;利用细胞因子ELISA试剂盒检测不同浓度CDPK9处理48h对Ana-1巨噬细胞TNF-α、TGF-β1、IL-10、IL-1β分泌的影响。结果如下:CDPK9能够与Ana-1巨噬细胞结合。CDPK9对Ana-1巨噬细胞增殖具有抑制作用,较高浓度时抑制作用明显。CDPK9能够显著提高Ana-1巨噬细胞的凋亡比例,同时促进其吞噬能力。CDPK9上调了Ana-1巨噬细胞NO产量。细胞因子IL-1β和TNF-α的表达量上升,而TGF-β1和IL-10的表达量有所下降。CDPK9能够与巨噬细胞结合并通过多种途径影响其活性及免疫功能。This study aimed to investigate regulation of calcium dependent protein kinase 9 of Toxoplasma gondii to the macrophages of mouse. The laser confocal microscopy was utilized to check whether CDPK9 was able to bind to Ana 1 macrophages. After co-incubation with CDPKg, the proliferation of macrophages was checked by cell counting kit-8 （CCK-8）. Then apoptosis and phagocytosis of macrophages were calculated by flow eytometry. NO production of cells was eval- uated by Total Nitric Oxide Assay kit and expressions of TNF-α, TGF-β1, IL 10 and IL-1β were detected by ELISA research reagent. The pictures of laser confocal microscopy showed that CD- PK9 could bind to macrophages. After co-incubation with CDPK9, the proliferation of macropha- ges was suppressed. Moreover, both apoptosis and phagocytosis rates of cells were increased. However, NO production was up-regulated by CDPK9. Concerning cytokines, production of IL- 1β and TNF-αwas increased, while the production of TGF-β1 and IL-10 was decreased. Our study demonstrated that CDPK9 was able to bind to macrophages and exert its effects on the activity and functions of Ana-1 macrophages via different pathways.

关 键 词：刚地弓形虫 钙依赖蛋白激酶9 巨噬细胞 流式细胞术 细胞因子 

分 类 号：S855.9[农业科学—临床兽医学] S852.72[农业科学—兽医学]