CAJ | 학술논문

目的：观察内吗啡肽-1（EM-1）对人外周血来源树突状细胞TLR2和TLR4表达的影响。方法从人外周血中提取和分离单核细胞，在含重组人粒-巨噬细胞集落刺激因子和重组人白细胞介素-4的完全培养基中培养，6d后未成熟树突状细胞（imDC）分4组，空白对照组（BLA组）、EM-1组、LPS组、LPS＋EM-1组。继续培养2 d后，流式细胞术检测各组DC表面 TLR2、TLR4蛋白的表达；RT-PCR 法检测各组DC中TLR2 mRNA、TLR4 mRNA的表达。结果流式结果显示，TLR2、TLR 4在imDC表达较高，随着DC成熟表达下降。与BLA组相比，EM-1组DC表面TLR2、TLR4的表达下调（P＜0.05）；与LPS组相比，LPS＋EM-1组DC表面TLR2、TLR4受体均明显下调（P＜0.01）。RT-PCR的结果显示，与BLA组相比，EM-1干预后引起DC 表面TLR2 mRNA表达明显降低（P＜0.01），TLR4 mRNA的变化无差异（P＞0.05）；与LPS组相比，LPS ＋EM-1组 DC 表面 TLR2 mRNA、TLR4 mRNA均有所下调（P ＜0.05）。结论 EM-1使 DC 表面TLR2、TLR4的表达下调，EM-1对DC免疫功能的影响可能与DC表面TLR2、TLR4表达有关。
목적：관찰내마배태-1（EM-1）대인외주혈래원수돌상세포TLR2화TLR4표체적영향。방법종인외주혈중제취화분리단핵세포，재함중조인립-거서세포집락자격인자화중조인백세포개소-4적완전배양기중배양，6d후미성숙수돌상세포（imDC）분4조，공백대조조（BLA조）、EM-1조、LPS조、LPS＋EM-1조。계속배양2 d후，류식세포술검측각조DC표면 TLR2、TLR4단백적표체；RT-PCR 법검측각조DC중TLR2 mRNA、TLR4 mRNA적표체。결과류식결과현시，TLR2、TLR 4재imDC표체교고，수착DC성숙표체하강。여BLA조상비，EM-1조DC표면TLR2、TLR4적표체하조（P＜0.05）；여LPS조상비，LPS＋EM-1조DC표면TLR2、TLR4수체균명현하조（P＜0.01）。RT-PCR적결과현시，여BLA조상비，EM-1간예후인기DC 표면TLR2 mRNA표체명현강저（P＜0.01），TLR4 mRNA적변화무차이（P＞0.05）；여LPS조상비，LPS ＋EM-1조 DC 표면 TLR2 mRNA、TLR4 mRNA균유소하조（P ＜0.05）。결론 EM-1사 DC 표면TLR2、TLR4적표체하조，EM-1대DC면역공능적영향가능여DC표면TLR2、TLR4표체유관。
Aim To observe the effect of endomorphin-1 (EM-1 )on TLR2 and TLR4 expressions of dendritic cells (DC)from human peripheral blood.Methods Monocytes isolated from human peripheral blood mono-nuclear cells were cultured in medium containing re-combinant human interleukin-4 and recombinant hu-man granulocyte macrophage colony stimulating factor. After six days of culture,the immature dendritic cells (imDC ) were divided into four groups,the control group (BLA group),EM-1 group,LPS group and LPS+EM-1 group.After 2 days of culture,the expres-sions of TLR2 and TLR4 were determined by fluores-cence activated cell sorter(FACS).The expressions of TLR2 and TLR4 at mRNA level in DC were detected by RT-PCR.Results The FACS results showed that the expressions of TLR2 and TLR4 in imDC were high-er,and their expressions were decreased with the mat-uration of DC.Compared with BLA group,the expres-sions of TLR2 and TLR4 in DC were down-regulated in EM-1 group (P<0.05 ).Compared with LPS group, TLR2,TLR4 on DC surface were significantly lower in LPS +EM-1 group (P <0.01 ). RT-PCR results showed that compared with BLA group,EM-1 interven-tion induced TLR2 mRNA expression was down-regula-ted significantly (P<0.01),there were no significant changes of TLR4 mRNA expression (P>0.05 ).mR-NA expressions of TLR2 and TLR4 on DC in LPS +EM-1 group were lower than those in LPS group (P<0.05 ).Conclusions EM-1 enables the down-regula-tion of the expressions of TLR2 and TLR4 on DC sur-face,the effects of EM-1 on immune function may be associated with TLR2 and TLR4 expressions on DC surface.