人单核细胞系THP-1细胞经刚地弓形虫感染产生IL-12的机制研究  

Research on the action mechanism of toxoplasma gondii-induced IL-12 production in human THP-1 cells

作　　者：刘红珍[1] 李文建[2] 仝宇红 钱雪松[2] LIU Hongzhen;LI Wenjian;TONG Yuhong(Department of Protistology,Medical Department of Hebei Engineering University,Hebei,Handan 056000,Chin)

机构地区：[1]河北工程大学医学部病原生物学教研室,河北省邯郸市056000 [2]河北医科大学免疫教研室河北省重大疾病的免疫机制及干预重点实验室 [3]河北省石家庄市第一医院感染管理控制处

出　　处：《河北医药》2018年第12期1765-1768,1773,共5页Hebei Medical Journal

基　　金：河北省邯郸市科学技术研究与发展计划项目(编号:1423108064-9)

摘　　要：目的探讨刚地弓形虫感染人单核细胞系THP-1细胞产生IL-12机制。方法培养人单核细胞系THP-1细胞株,分为对照组和感染组。通过ELISA法检测2组细胞培养上清中IL-12的浓度;实时定量PCR检测2组中IL-12 p35和IL-12 p40亚基m RNA表达情况;构建不同长度IL-12 p35启动子片段并转染到RAW264.7细胞株中,采用荧光检测仪检测荧光强度。结果细胞因子IL-12在弓形虫感染THP-1细胞后,细胞培养上清中分泌明显增加;实时定量PCR检测IL-12 p35和IL-12 p40 m RNA表达,与对照组相比,IL-12 p35及IL-12 p40 m RNA表达明显升高,差异有统计学意义(P<0.05)。成功构建3种不同长度启动子质粒后,经转染处理后,与对照组相比,p35(-1 080~1 bp)/Px P2、p35(-1 080^-20 bp)/Px P2质粒荧光素酶活性增加,差异有统计学意义(P<0.05),而p35(-1 080^-50 bp)/Px P2质粒荧光素酶活性无差异。结论弓形虫感染单核细胞THP-1后,引起IL-12分泌的增加,可能是作用于IL-12启动子-20^-50这段核心区域所引起的。Objective To investigate the action mechanism of toxoplasma gondii-induced IL-12 production in human THP-1 cells. Methods The cultured human monocytic cells series THP-1 cells were divided into control group and infection group. The THP-1 cells in infection group were stimulated with Toxoplasma gondii,then the levels of cytokine-IL-12 in cell culture supernatant were detected in both groups by ELISA,moreover,the expression levels of IL-12 p35 and p40 gene m RNA were measured in both groups by real-time quantitative PCR（ RT-PCR）. The three different length of IL-12 promoter region of plasmids were successfully constructed,which were transfected into RAW264. 7 cells,and the fluorescence intensity was detected by fluorescence detector. Results The levels of IL-12 in culture supernatant in infection group were increased significantly. As compared with those in control group,the expression levels of p35 and p40 gene m RNA in infection group were significantly increased（ P〈 0. 05）. The three different length of IL-12 promoter region of plasmids were successfully constructed,which were transfected into RAW264. 7 cells. As compared with that in control group,the fluorescence intensities of p35（-1,080 - 1 bp）/Px P2 and p35（-1,080 --20 bp）/Px P2 of plasmids were significantly increased（ P〈 0. 05）,however,there was no significant difference in p35（-1,080 --50 bp）/Px P2 plasmid luciferase activity between two groups.Conclusion The levels of IL-12 in THP-1 cells after stimulated with Toxoplasma gondii are increased significantly,which may be caused by the-20 --50 core area of IL-12 promoter.

关 键 词：弓形虫 单核细胞 白介素12 启动子 

分 类 号：R53[医药卫生—内科学]