人绒毛外滋养层细胞HTR-8/SVneo中Toll样受体mRNA的表达及对吲哚胺2,3-双加氧酶mRNA水平的影响

Effects of Toll- like receptors on indoleamine 2, 3- dioxygenase mRNA levels in human trophoblast HTR-8/SVneo cells

目的系统检测人绒毛外滋养细胞HTR-8/SVneo中Toll样受体(TLR)mRNA的表达情况,定量分析TLRs配体刺激前后吲哚胺2,3双加氧酶(indoleamine 2,3-dioxygenase,IDO)mRNA的表达差异。方法 RT-PCR检测HTR-8/SVneo细胞中TLR 1-10mRNA的表达情况;实时荧光定量RT-PCR检测TLR3、TLR4、TLR7/8、TLR9配体刺激前后IDO mRNA表达水平。结果 HTR-8/SVneo细胞中有IDO及TLR 1-10 mRNA表达;在48 h内IDO mRNA表达随着时间延长,呈升高趋势;IDO mRNA的表达与培养基营养状况相关;TLRs配体刺激后,除TLR-3转录水平表达下调外,TLR-4、7、8、9 mRNA表达均上调;poly(I∶C)刺激后IDO mRNA表达低于正常组,IFN-γ刺激后表达高于正常组(P<0.05)。结论 TLRs mRNA的表达提示该细胞具有抗感染作用;HTR-8/Svneo细胞中IDO mRNA的表达与母胎界面营养状况相关;TLRs配体刺激剂对TLRs及IDO mRNA表达的影响不仅验证了TLRs的功能活性,而且提示了IDO在抗病原体免疫应激中可依赖TLRs途径发挥作用。

Objective To study the expression of Toll-like receptors （TLRs） mRNA in human trophoblast HTR-8/SVneo cells and the changes in indoleamine 2,3-dioxygenase （IDO） mRNA expression in response to TLR ligand stimulation. Methods The expressions of TLRs and IDO mRNA in human HTR-8/SVneo ceils were tested by RT-PCR, and the changes in IDO mRNA levels after exposure to TLR3, TLR4, TLR7/8, and TLR9 ligands were quantitatively analyzed with real-time PCR. Results IDO and TLRI-10 mRNAs were expressed in HTR-8/SVneo cells. As the cell culture time extended, IDO mRNA expression level tended to increase within 48 h. After stimulation with the TLR ligands, the expression of TLR-3 mRNA was down-regulated while the expression of TLR-4, 7, 8, and 9 mRNA up-regulated. Stimulation of the ceils with poly（I：C） lowered the expression of IDO mRNA while IFN-y increased its expression. Conclusions The expression of 1DO mRNA is associated with the nutrition of the maternal-fetal interface. Stimulation with the TLR ligands affects the expression of IDO and TLR mRNA expressions in the cells, which verifies the functional activity of TLRs and suggests a role of IDO in TLR pathway-dependent antiviral immunity.