polyI:C诱导人胎盘滋养层细胞抗HBV感染的体外研究  

In vitro study on anti-HBV effect of human placental trophoblast cells induced by PolyI:C

作　　者：资捷 王前[2] 郑磊[2] 熊石龙[2] 王芳 

机构地区：[1]广东省深圳市福田妇幼保健院,518026 [2]南方医科大学南方医院临床检验中心

出　　处：《中国妇幼保健》2012年第8期1228-1231,共4页Maternal and Child Health Care of China

摘　　要：目的:探讨体外polyI:C对人胎盘滋养层细胞乙型肝炎病毒(HBV)感染的作用。方法:将2μg或8μg HBV重组质粒pcDNA3.1(+)-HBV1.3转染人胎盘滋养层细胞Bewo,12 h后以Toll样受体3(TLR3)配体polyI:C处理3天,同时设对照组。采用微粒子酶免疫分析法(MEIA)和荧光定量PCR法分别检测细胞上清液HBsAg、HBeAg及HBV DNA水平,并以荧光定量RT-PCR和ELISA分别检测细胞TLR3 mRNA表达及细胞上清IFN-β水平。结果:与对照组比较,polyI:C均可显著抑制Bewo细胞HBV复制(P<0.01或0.05),但与2μg HBV重组质粒转染组比较,转染8μg HBV重组质粒组polyI:C对HBV复制的抑制率显著下降(P<0.01),且polyI:C诱导Bewo细胞TLR3 mRNA表达及IFN-β水平显著下降,差异有统计学意义(P<0.05)。结论:polyI:C可诱导人胎盘滋养层细胞抗HBV感染,但随着HBV感染量的增大,通过下调细胞TLR3 mR-NA表达及IFN-β产生,其抗HBV感染的作用被显著削弱。Objective： To explore the effect of polyI： C on hepatitis B virus （HBV） infection in human placental trophoblast cells in vitro. Methods： HBV recombinant vector pcDNA3.1 （ ＋ ） - HBV1.3 of 2 μg or 8 μg were transfected into human placental trophoblast cells Bewo, after 12 hours, they were treated with Toll -like receptor （TLR3） ligand polyI： C for three days, control group was designed. MEIA and fluorescence quantitative PCR were used to detect the levels of HBsAg, HBeAg, and HBV DNA in liquid superuatant, fluorescence quantitative RT - PCR and ELISA were used to detect the expression of TLR3 mRNA and the level of IFN - β in liquid supernatant, respectively. Results： Compared with control group, polyI： C could suppressed HBV replication in Bewo cells significantly （P 〈0. 001 or 0. 05 ）, but compared with 2 μg HBV recombinant vector transfection group, the inhibiting rate of polyI： C to HBV replication decreased significantly in 8 μg HBV recombinant vector transfection group （ P 〈 0.01 ） . The expression of TLR3 mRNA and the level of IFN - β in Bewo cells induced by PolyI： C decreased significantly, there was significant difference （P 〈 0. 05 ） . Conclusion： PolyI： C can induce human placental trophoblast ceils against HBV infection, but with the increase of HBV infective dose, the anti - HBV infectious effect is reduced significantly by down - regulating the expression of TLR3 mRNA and the level of IFN - β.

关 键 词：polyI:C TOLL样受体3 胎盘 滋养层细胞 乙型肝炎病毒 

分 类 号：R512.62[医药卫生—内科学]