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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 detect the expression of Toll-like receptor (TLR) mRNA in human chorionic villus trophoblast cells HTR-8 / SVneo and to quantitatively analyze the expression of indoleamine 2,3-dioxygenase , IDO) mRNA expression differences. Methods The expression of TLR 1-10 mRNA in HTR-8 / SVneo cells was detected by RT-PCR. The expression of IDO mRNA in TLR3, TLR4, TLR7 / 8 and TLR9 ligands was detected by real-time fluorescence quantitative RT-PCR. Results IDO and TLR 1-10 mRNA expressions were observed in HTR-8 / SVneo cells. The expression of IDO mRNA increased with time in HTR-8 / SVneo cells. The expression of IDO mRNA was correlated with the nutritional status of media. TLRs ligand stimulation , The expressions of TLR-4, 7, 8 and 9 mRNA were up-regulated except for the down-regulation of TLR-3 transcription. The expression of IDO mRNA was lower in the stimulation of poly (I: C) Normal group (P <0.05). Conclusion The expression of TLRs mRNA suggests that the cells have anti-infective effect. The expression of IDO mRNA in HTR-8 / Svneo cells is related to the nutritional status of maternal-fetal interface. The effect of TLRs ligand agonists on the expression of TLRs and IDO mRNA not only verifies the function of TLRs Activity, but also suggests that IDO plays a role in TLRs-dependent pathogenic immune response.