CAJ | 학술논문

目的：探讨血管紧张素II（Ang II）对人单核细胞源树突状细胞（DCs）免疫成熟进程及摄取氧化低密度脂蛋白（ Ox-LDL）功能的影响。方法：采用密度梯度离心结合免疫磁珠法分离人外周血CD14＋单核细胞，经含100μg/L重组人粒细胞-巨噬细胞集落刺激因子（rhGM-CSF）和50μg/L重组人IL-4（rhIL-4）的RPMI-1640培养液培养，诱导分化为未成熟DCs。经不同浓度的Ang II干预24 h，另用10μmol/L血管紧张素II受体1拮抗剂洛沙坦预处理后再加入Ang II干预，采用流式细胞术检测细胞免疫表型（CD83和HLA-DR），酶联免疫吸附试验（ELISA）法检测细胞培养上清液中细胞因子（ IL-12和IFN-γ）的浓度；通过加入10 mg/L的荧光DiI标记的Ox-LDL，用流式细胞仪检测DiI-Ox-LDL的摄取分数；采用real-time PCR法检测清道夫受体A（SR-A）、CD36和凝集素样氧化低密度脂蛋白受体1（LOX-1） mRNA的表达。结果：（1） Ang II干预可上调DCs表面成熟标志物CD83和HLA-DR的表达，且分泌细胞因子IL-12和IFN-γ水平升高，10μmol/L洛沙坦预处理可明显抑制Ang II诱导的DCs 成熟。（2） Ang II干预可促进DCs对DiI-Ox-LDL的摄取，洛沙坦预处理可部分抑制DCs对DiI-Ox-LDL的摄取。（3） Ang II干预可上调LOX-1 mRNA的表达，但对SR-A和CD36的表达无明显影响，洛沙坦预处理可明显抑制LOX-1mRNA的表达，但SR-A和CD36的表达无明显变化。结论：Ang II可促进DCs的免疫成熟，并且可能通过上调LOX-1的表达，促进DCs对Ox-LDL的摄取，这可能是Ang II促动脉粥样硬化的新机制。
목적：탐토혈관긴장소II（Ang II）대인단핵세포원수돌상세포（DCs）면역성숙진정급섭취양화저밀도지단백（ Ox-LDL）공능적영향。방법：채용밀도제도리심결합면역자주법분리인외주혈CD14＋단핵세포，경함100μg/L중조인립세포-거서세포집락자격인자（rhGM-CSF）화50μg/L중조인IL-4（rhIL-4）적RPMI-1640배양액배양，유도분화위미성숙DCs。경불동농도적Ang II간예24 h，령용10μmol/L혈관긴장소II수체1길항제락사탄예처리후재가입Ang II간예，채용류식세포술검측세포면역표형（CD83화HLA-DR），매련면역흡부시험（ELISA）법검측세포배양상청액중세포인자（ IL-12화IFN-γ）적농도；통과가입10 mg/L적형광DiI표기적Ox-LDL，용류식세포의검측DiI-Ox-LDL적섭취분수；채용real-time PCR법검측청도부수체A（SR-A）、CD36화응집소양양화저밀도지단백수체1（LOX-1） mRNA적표체。결과：（1） Ang II간예가상조DCs표면성숙표지물CD83화HLA-DR적표체，차분비세포인자IL-12화IFN-γ수평승고，10μmol/L락사탄예처리가명현억제Ang II유도적DCs 성숙。（2） Ang II간예가촉진DCs대DiI-Ox-LDL적섭취，락사탄예처리가부분억제DCs대DiI-Ox-LDL적섭취。（3） Ang II간예가상조LOX-1 mRNA적표체，단대SR-A화CD36적표체무명현영향，락사탄예처리가명현억제LOX-1mRNA적표체，단SR-A화CD36적표체무명현변화。결론：Ang II가촉진DCs적면역성숙，병차가능통과상조LOX-1적표체，촉진DCs대Ox-LDL적섭취，저가능시Ang II촉동맥죽양경화적신궤제。
AIM:To investigate the effects of angiotensin II ( Ang II) on the immune maturation and the oxi-dized low-density lipoprotein (Ox-LDL)-uptaking capacity of human monocyte-derived dendritic cells (DCs).METH-ODS:Human peripheral blood mononuclear cells were isolated by density gradient centrifugation , and the monocytes were purified by positive selection with anti-CD14 magnetic beads.After cultured with rhGM-CSF (100 μg/L) and rhIL-4 (50μg/L) for 5 d, the monocytes differentiated into immature DCs .On the 6th day of the culture, the cells were treated with various concentration levels of Ang II or pretreated with losartan .The immunophenotypic expression of HLA-DR and CD83 was analyzed by flow cytometry .The secretion levels of IL-12 and IFN-γin the culture supernatants were measured by ELISA.Furthermore, DCs were incubated with DiI-labelled Ox-LDL.The DiI-Ox-LDL-incorporated fraction was investiga-ted by flow cytometry .The mRNA expression of 3 scavenger receptors , scavenger receptor A ( SR-A) , CD36 and lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1), was examined by real-time PCR.RESULTS: Ang II induced the maturation of human monocyte-derived DCs, stimulated the expression of CD83 and HLA-DR, and promoted the secre-tion of IL-12 and IFN-γ, which were suppressed by losartan .Furthermore, Ang II increased the Ox-LDL-uptaking capacity of DCs, which was partially reduced by losartan .The incubation of DCs with Ang II enhanced the mRNA expression of LOX-1 in a dose-dependent manner , which was reduced by losartan .However, the expression of SR-A and CD36 was not changed .CONCLUSION:Ang II promotes the immune maturation of human monocyte-derived DCs and increases the up-take of Ox-LDL probably through the up-regulation of LOX-1 expression.