人白细胞介素.doc

HepG2.2.15细胞株中TLR7激活诱导的炎症因子表达 张胜权, 胡道军, 朱娟娟, 罗 欣, 黄海量, 柴 瑜, 陶千山 （安徽医科大学生物化学与分子生物学教研室，安徽 合肥230032） 摘要： 目的 探讨乙型肝炎病毒(HBV)感染是否诱导HepG2.2.15中TLR7表达，分析HepG2.2.15细胞中TLR7激活后诱导炎症因子表达。方法 通过荧光定量PCR分析HepG2和HBV转染细胞株HepG2.2.15中TLR7受体mRNA水平表达。TLR7配体Gardiquimod刺激20 min后，Western-blot分析激活促信号通路；刺激6 h和48 h后，荧光定量PCR分析TLR7激活后IL-6 和TNF-αmRNA水平表达变化；刺激48 h后，ELISA法分析IL-6 和TNF-α蛋白质水平表达变化。结果 HBV转染的HepG2.2.15细胞株中TLR7 mRNA水平表达比HepG2中的表达高约30倍（P0.01)。Gardiquimod刺激6 h后，IL-6 和TNF-αmRNA表达水平明显增加（P0.01)；刺激48 h后，IL-6 和TNF-α蛋白质表达水平明显增加（P0.01)。HepG2.2.15细胞株经Gardiquimod刺激后NF-κB p65亚单位进入细胞核中量显著增加；结论 HBV感染能够诱导HepG2.2.15细胞中TLR7表达，TLR7激活后能够上调细胞中促炎症因子的表达，提示TLR7可能在乙型病毒性肝炎的病理生理过程中发挥作用 关键词：乙型肝炎病毒；TLR7； Gardiquimod; HepG2.2.15细胞株; [中图分类号]R392.11 [文献标识码] Expression of the Inflammation Factors Induced by the Activation of TLR7 in HepG2.2.15 Ce ZHANG Sheng-Quan Hu Dao-Jun Zhu Juan-Jjuan LUO Xin Huang Hai-Liang Chai Yu Tao Qian-Shan Department of Biochemistry and Molecular Biology, Anhui Medical University, Hefei 230032, China Abstract： Objective To explore whether the infection of hepatitis B virus can induce the expression of TLR7 in HepG2.2.15 基金项目：安徽省自然科学基金资助（050430604）及安徽省重点实验室优秀中青年科研带头人专项基金资助（安徽省科技厅，2007年）。 作者简介：张胜权,男，博士，副教授,硕士生导师，从事免疫分子生物学研究，发表论文39篇Tel:0551-5161194, Email: sqz36@. TLR7-induced inflammation factors in HepG2.2.15 cell line. Methods We performed a Real-time PCR to analyze the difference expression of TLR7 between in HepG2 and HepG2.2.15 cell lines. The messenger ribonucleic acid(mRNA) expression of TNF-α and IL-6 was measured by Real-time PCR and the protein secretion of the two cytokines was determined by ELISA method. Western blot was used to analyze the activation of NF-κB in HepG2.2.15 cell lines. Results The expression of TLR7 in HepG2.2.15 cells was about 30-fold higher than the one in the HepG2 cells. After HepG2.2.15 cells was stimulated by Gardiquimod , the mRNA and the protein levels of IL-6 and TNF-α significantly increased about（compare with the control P0.01)