CAJ | 학술논문

目的观察阻断Th17细胞分化对小鼠小肠急性移植物抗宿主病(aGVHD)的影响.方法将小鼠分为5组:正常对照组、单纯照射组、异基因骨髓移植(allo-BMT)组、allo-BMT+二甲基亚砜(DMSO)组、allo-BMT+常山酮(HF)组.移植后观察小鼠GVHD发生情况并进行临床和病理评分;移植后第14天(+14 d),用流式细胞术检测外周血Th1、Th17比例及IFN-γ和IL-17A浓度变化;+14 d,用RT-PCR方法检测小肠T-bet、RORγT、CXCL9、CXCL10、CXCL11和CCL20转录因子表达水平.结果 +14 d,allo-BMT+HF组小鼠小肠损伤较重;allo-BMT+HF组Th1细胞比例高于allo-BMT组[分别为(9.34±0.67)%和(6.69±1.37)%],Th17细胞明显下降[分别为(0.46±0.04)%和(1.01±0.04)%](P<0.05),allo-BMT组IL-17A和IFN-γ浓度分别为(3.54±0.06)和(41.67±0.11)ng/L,而allo-BMT+HF组IFN-γ浓度为(38.77±0.17)ng/L(P<0.05),未检测到IL-17A.RT-PCR分析结果显示allo-BMT+HF组小鼠小肠组织中RORγT表达均较allo-BMT+BMT和正常对照组低(P<0.01);allo-BMT和allo-BMT+HF组T-bet表达量均较正常对照组表达升高(P值均<0.05),但二者相比差异无统计学意义(P>0.05);移植后allo-BMT-HF组CXCL9、CXCL10表达明显高于allo-BMT组(P<0.01),但未检测到CCL20.结论阻断Th17细胞分化加重allo-BMT小鼠小肠损伤.
목적 관찰조단Th17세포분화대소서소장급성이식물항숙주병(aGVHD)적영향.방법 장소서분위5조:정상대조조、단순조사조、이기인골수이식(allo-BMT)조、allo-BMT+이갑기아풍(DMSO)조、allo-BMT+상산동(HF)조.이식후관찰소서GVHD발생정황병진행림상화병리평분;이식후제14천(+14 d),용류식세포술검측외주혈Th1、Th17비례급IFN-γ화IL-17A농도변화;+14 d,용RT-PCR방법검측소장T-bet、RORγT、CXCL9、CXCL10、CXCL11화CCL20전록인자표체수평.결과 +14 d,allo-BMT+HF조소서소장손상교중;allo-BMT+HF조Th1세포비례고우allo-BMT조[분별위(9.34±0.67)%화(6.69±1.37)%],Th17세포명현하강[분별위(0.46±0.04)%화(1.01±0.04)%](P<0.05),allo-BMT조IL-17A화IFN-γ농도분별위(3.54±0.06)화(41.67±0.11)ng/L,이allo-BMT+HF조IFN-γ농도위(38.77±0.17)ng/L(P<0.05),미검측도IL-17A.RT-PCR분석결과현시allo-BMT+HF조소서소장조직중RORγT표체균교allo-BMT+BMT화정상대조조저(P<0.01);allo-BMT화allo-BMT+HF조T-bet표체량균교정상대조조표체승고(P치균<0.05),단이자상비차이무통계학의의(P>0.05);이식후allo-BMT-HF조CXCL9、CXCL10표체명현고우allo-BMT조(P<0.01),단미검측도CCL20.결론 조단Th17세포분화가중allo-BMT소서소장손상.
Objective To study the role of Th17 cells in acute intestine graft-versus-host disease following allogenetic bone marrow transplantation(allo-BMT). Methods Mice were split randomly into five groups: normal control, irradiated, allo-BMT, allo-BMT+DMSO and allo-BMT+ Halofuginone (HF) groups . HF was given intraperitoneally at a dose of 5 μg per mouse from -1 d to 10 d after allo-BMT. aGVHD symptoms were followed-up to perform clinical and pathogenic scores. The levels of Th1/Th17, interleukin-17 and interferon-γ were measured by flow cytometry at day 7 d. mRNA expressions of T-bet, RORγT, CXCL9, CXCL10, CXCL11 and CCL20 in intestine were evaluated by real-time PCR. Results Intestinal damages in allo-BMT-HF mice was more serious than in normal control and allo-BMT groups at day 14 after transplantation. At day 7, Th17 ratio in allo-BMT+HF group was significantly lower than in allo-BMT group. IL-17A was not detected, but Th1 ratio was higher in allo-BMT+HF. There was a similar increment in the relative expressions of T-bet in both allo-BMT and allo-BMT+HF groups. Expressions of CXCL9 and CXCL10 elevated in allo-BMT+HF group, which were significantly higher than those in allo-BMT group (P<0.01). CCL20 expression significantly increased in allo-BMT group, but it was not detected in allo-BMT+HF group. Conclusion Blockage of th17 cells differentiation exacerbated acute intestine graft versus-host disease.