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

doi:10.11843/j.issn.0366-6964.2017.04.017

摘要/Abstract

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24 h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48 h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48 h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48 h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。

Abstract:

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 CDPK9, the proliferation of macrophages was checked by cell counting kit-8 (CCK-8). Then apoptosis and phagocytosis of macrophages were calculated by flow cytometry. NO production of cells was evaluated 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 CDPK9 could bind to macrophages. After co-incubation with CDPK9, the proliferation of macrophages 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-α was 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.

中图分类号:

马群山, 刘欣超, 孙晓妮, 王帅, 徐立新, 宋小凯, 严若峰, 李祥瑞. 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[J]. 畜牧兽医学报, 2017, 48(4): 731-739.

MA Qun-shan, LIU Xin-chao, SUN Xiao-ni, WANG Shuai, XU Li-xin, SONG Xiao-kai, YAN Ruo-feng, LI Xiang-rui. Effects of Calcium Dependent Protein Kinase 9 of Toxoplasma gondii on the Functions of Mouse Ana-1 Macrophages in vitro[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(4): 731-739.

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