HCV感染诱导BV-2细胞TNF-α分泌与TLR2 mRNA表达的关系  

The relationship between TNF-α secretion and TLR2 mRNA expression in HCV infected BV-2 cells

作　　者：杨娟[1] 王国玮[1] 王振海[1] 

机构地区：[1]宁夏医科大学附属医院神经内科,750004

出　　处：《中国神经免疫学和神经病学杂志》2011年第1期20-23,共4页Chinese Journal of Neuroimmunology and Neurology

基　　金：国家自然科学基金资助项目(30760067);教育部春晖计划基金资助项目(z-2008-1-7502);宁夏医科大学博士点建设开发课题资助项目

摘　　要：目的探讨丙型肝炎病毒(HCV)感染后小鼠小胶质细胞(BV-2)上清液中TNF-α水平与Toll样受体2(TLR2)mRNA表达的相关性,为进一步探讨TLR2介导的小胶质细胞抗HCV的免疫机制提供依据。方法将BV-2细胞分为空白对照组(常规培养用DMEM高糖培养液)、正常血清对照组〔常规培养用含有20%(体积分数)正常血清的DMEM高糖培养液〕及实验组〔常规培养用含有20%(体积分数)HCV阳性血清的DMEM高糖培养液〕,分别以ELISA方法检测各组HCV感染后3、6、9 h的BV-2细胞培养上清液中的TNF-α水平,以RT-PCR法检测同组相同时间点BV-2细胞TLR2 mRNA表达变化,并分析两者的相关性。结果空白对照组与正常血清对照组相同时间点TLR2 mRNA表达及TNF-α水平比较无统计学差异(P>0.05)。实验组3、6、9h TNF-α水平分别为(144.67±5.51)(、168.33±3.06)、(185.50±4.82)pg/mL,均高于两对照组〔空白对照组分别为(43.17±1.79)(、44.23±2.13)、(45.61±2.38)pg/mL、正常血清对照组分别为(43.33±1.97)、(43.34±1.55)(、48.08±2.24)pg/mL〕(均P<0.05);实验组3、69、h时TLR2 mRNA表达水平分别为1.08±0.061、.42±0.05、1.61±0.57,各时间点间比较有统计学差异(P<0.05),实验组69、h与3 h比较有统计学差异(P<0.05),但9 h时与6 h时比较无统计学差异(P>0.05)。Pearson相关分析显示TLR2 mRNA表达与TNF-α分泌水平呈正相关(r=0.819,P<0.01)。结论 HCV感染后TLR2表达及TNF-α水平均明显增高,两者可能参与了BV-2细胞抗HCV的免疫过程。Objective To detected the expression of TLR2 mRNA and the levels of TNF-α in supernatant of HCV infected microglial cells in mice （BV-2 cell）, and to explore the immune mechanisms against HCV mediated by TLR2 in microglia. Methods BV-2 ceils were divided into the control group, the normal serum control group that cultured with 20% normal serum containing DMEM high glucose medium and the experimental group that cultured with 20% HCV-positive serum DMEM high glucose medium. The expression of TLR2 mRNA was detected by RT-pCR, and the level of TNF-α was measured with ELISA at 3 h, 6 h, 9 h after HCV infection. The correlation between these two factors was analyzed by Pearson linear correlation analysis. Results There was no significant difference of the levels of TLR2 mRNA between the control group and the normal serum control group （P〉0.05）. The TNF-α levels at 3 h, 6 h, 9 h in the experimental group [（144. 67±5.51）, （168.33±3.06）, （185.50± 4.82） pg/mL, respectively] were higher than in the control groups [ （43.17±1.79）, （44.23±2.13）, （45.61±2.38） pg/mL, respectively] （P〈0. 05）, and it showed a gradually increasing trend. The expression of TLR2 mRNA after infection appeared significantly increase at 3 h, peaked at 6 h and there was significant difference between 3 h, and 6 h, 9 h （1.08±0. 06, 1.42±0.05, 1.61± 0.57, respectively） in the experimental group （P〈0.05）, but there was no significant difference between 6 h and 9 h groups （P〉0.05）. Furthermore, expression of TLR2 mRNA and level of TNF-α was positively correlated with a correlation coefficient of 0. 819 （P〈0.01） showed by Pearson correlation analysis. Conclusions The expression of TLR2 mRNA and level of TNF-α significantly increased in BV-2 cells after HCV infection, both of which might be involved in anti-HCV immunity.

关 键 词：丙型肝炎病毒 小鼠小胶质细胞 TOLL样受体2 肿瘤坏死因子-α 

分 类 号：R512.3[医药卫生—内科学]