CAJ | 학술논문

目的：检测小鼠肠系膜淋巴结树突状细胞（ MLNDC）上促肾上腺皮质激素释放因子（CRF）及其受体CRFR1、CRFR2的表达，探讨CRF及其受体对MLNDC的表型改变的影响。方法采用磁珠分选技术分离C57BL／6小鼠的MLNDC，以流式标记CD11c鉴定DC分选纯度，获得高纯度MLNDC后，RT-PCR检测其CRF、CRFR1与CRFR2的转录水平，免疫荧光双染法明确MLNDC表面CRFR1及CRFR2的表达。将MLNDC体外培养，予以CRF受体相应拮抗剂干预，流式细胞术观察DC表面分子变化。结果流式细胞术检测显示经磁珠分选后， CD11 c标记的树突状细胞阳性率为（80．12±6．34）％，锥虫蓝染色显示细胞活性＞90％，MLNDC存在CRF、CRFR1与CRFR2 mRNA的表达，免疫荧光提示其表面存在CRFR1及CRFR2的表达，CRFR1拮抗剂Antalarmin（10 nmol／L）可以下调MLNDC表面共刺激分子CD86，而CRFR2拮抗剂Astressin 2B（10 nmol／L）可以上调MLNDC表面共刺激分子CD86。结论体外分离的C57 BL／6小鼠MLNDC自身存在CRF 及其受体CRFR1和CRFR2的表达，并且该两种受体对MLNDC表型具有相反的作用。
목적：검측소서장계막림파결수돌상세포（ MLNDC）상촉신상선피질격소석방인자（CRF）급기수체CRFR1、CRFR2적표체，탐토CRF급기수체대MLNDC적표형개변적영향。방법채용자주분선기술분리C57BL／6소서적MLNDC，이류식표기CD11c감정DC분선순도，획득고순도MLNDC후，RT-PCR검측기CRF、CRFR1여CRFR2적전록수평，면역형광쌍염법명학MLNDC표면CRFR1급CRFR2적표체。장MLNDC체외배양，여이CRF수체상응길항제간예，류식세포술관찰DC표면분자변화。결과류식세포술검측현시경자주분선후， CD11 c표기적수돌상세포양성솔위（80．12±6．34）％，추충람염색현시세포활성＞90％，MLNDC존재CRF、CRFR1여CRFR2 mRNA적표체，면역형광제시기표면존재CRFR1급CRFR2적표체，CRFR1길항제Antalarmin（10 nmol／L）가이하조MLNDC표면공자격분자CD86，이CRFR2길항제Astressin 2B（10 nmol／L）가이상조MLNDC표면공자격분자CD86。결론체외분리적C57 BL／6소서MLNDC자신존재CRF 급기수체CRFR1화CRFR2적표체，병차해량충수체대MLNDC표형구유상반적작용。
Objective To investigate the expression of corticotropin-releasing factor (CRF) and its receptors including CRFR1 and CRFR2 on mouse mesenteric lymph nodes dendritic cells (MLNDC), and to analyze their effects on the biological phenotypes of intestinal dendritic cells .Methods The MLNDCs were isolated from C57BL/6 mice by using magnetic bead sorting .The purity of CD11c+DCs was identified by flow cytometry .The double-labeling immunofluorescence and the reverse transcriptase-polymerase chain reaction (RT-PCR) were performed to detect the expression of CRF , CRFR1 and CRFR2 on MLNDCs.The MLNDCs were exposed to CRF with or without the interference of CRFR 1 and CRFR2 antagonists .Flow cy-tometry was used to measure the changes of surface molecules ( MHCⅠ and MHCⅡ) and co-stimulatory molecules (CD80 and CD86).Results The CD11c+DCs accounted for (80.12±6.34)% of the isolate cells with a high cell viability of more than 90%.The expression of CRF , CRFR1 and CRFR2 at mRNA lev-el were detected in MLNDCs by RT-PCR.Results of the immunofluorescent staining assay indicated that both CRFR1 and CRFR2 were expressed on the surface of MLNDCs .The expression of CD86 on MLNDCs was inhibited by the treatment of MLNDCs with CRFR 1 antagonist , but enhanced by the treatment with CRFR2 antagonist .Conclusion Both CRF and CRFRs were detected in the MLNDCs isolated from the C57BL/6 mice.The CRF could alter the biological phenotypes of MLNDCs through binding to different CRFRs (CRFR1 and CRFR2), which affected the phenotypes of MLNDCs in opposite ways .