南方医科大学学报
南方醫科大學學報
남방의과대학학보
JOURNAL OF SOUTHERN MEDICAL UNIVERSITY
2013年
11期
1559-1564
,共6页
许巍%杨贵波%端家忠%王越%姚文荣%刘学庆%陈雪梅%丁裕斌%王应雄%何俊琳
許巍%楊貴波%耑傢忠%王越%姚文榮%劉學慶%陳雪梅%丁裕斌%王應雄%何俊琳
허외%양귀파%단가충%왕월%요문영%류학경%진설매%정유빈%왕응웅%하준림
HTR-8/SVneo%Toll样受体%吲哚胺2,3双加氧酶
HTR-8/SVneo%Toll樣受體%吲哚胺2,3雙加氧酶
HTR-8/SVneo%Toll양수체%신타알2,3쌍가양매
HTR-8/SVneo%Toll-like receptors%indoleamine 2,3-dioxygenase
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 1-10 mRNA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。
目的：繫統檢測人絨毛外滋養細胞HTR-8/SVneo中Toll樣受體（TLR）mRNA的錶達情況，定量分析TLRs配體刺激前後吲哚胺2,3雙加氧酶（indoleamine 2,3-dioxygenase, IDO）mRNA的錶達差異。方法 RT-PCR檢測HTR-8/SVneo細胞中TLR 1-10 mRNA的錶達情況；實時熒光定量RT-PCR檢測TLR3、TLR4、TLR7/8、TLR9配體刺激前後IDO mRNA錶達水平。結果 HTR-8/SVneo細胞中有IDO及TLR 1-10 mRNA錶達；在48 h內IDO mRNA錶達隨著時間延長，呈升高趨勢；IDO mRNA的錶達與培養基營養狀況相關；TLRs配體刺激後，除TLR-3轉錄水平錶達下調外，TLR-4、7、8、9 mRNA錶達均上調；poly（I∶C）刺激後IDO mRNA錶達低于正常組，IFN-γ刺激後錶達高于正常組（P<0.05）。結論 TLRs mRNA的錶達提示該細胞具有抗感染作用；HTR-8/Svneo細胞中IDO mRNA的錶達與母胎界麵營養狀況相關；TLRs配體刺激劑對TLRs及IDO mRNA錶達的影響不僅驗證瞭TLRs的功能活性，而且提示瞭IDO在抗病原體免疫應激中可依賴TLRs途徑髮揮作用。
목적：계통검측인융모외자양세포HTR-8/SVneo중Toll양수체（TLR）mRNA적표체정황，정량분석TLRs배체자격전후신타알2,3쌍가양매（indoleamine 2,3-dioxygenase, IDO）mRNA적표체차이。방법 RT-PCR검측HTR-8/SVneo세포중TLR 1-10 mRNA적표체정황；실시형광정량RT-PCR검측TLR3、TLR4、TLR7/8、TLR9배체자격전후IDO mRNA표체수평。결과 HTR-8/SVneo세포중유IDO급TLR 1-10 mRNA표체；재48 h내IDO mRNA표체수착시간연장，정승고추세；IDO mRNA적표체여배양기영양상황상관；TLRs배체자격후，제TLR-3전록수평표체하조외，TLR-4、7、8、9 mRNA표체균상조；poly（I∶C）자격후IDO mRNA표체저우정상조，IFN-γ자격후표체고우정상조（P<0.05）。결론 TLRs mRNA적표체제시해세포구유항감염작용；HTR-8/Svneo세포중IDO mRNA적표체여모태계면영양상황상관；TLRs배체자격제대TLRs급IDO mRNA표체적영향불부험증료TLRs적공능활성，이차제시료IDO재항병원체면역응격중가의뢰TLRs도경발휘작용。
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 cells 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 TLR1-10 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 cells with poly(I:C) lowered the expression of IDO mRNA while IFN-γincreased its expression. Conclusions The expression of IDO 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.
