CAJ | 학술논문

目的探讨大鼠肠道树突细胞(DC)介导肥大细胞活化对肠易激综合征(IBS)形成的作用机制.方法 20只SD大鼠按数字表随机分成对照组及模型组(各10只),模型组以结直肠扩张联合束缚应激建立大鼠内脏高敏感模型,以腹部收缩反射(AWR)实验评估大鼠内脏敏感性,免疫组织化学方法检测回盲部结肠组织表面分子CD103表达,甲苯胺蓝染色法计数回盲部结肠黏膜肥大细胞,酶联免疫吸附试验(E LISA)法检测肠黏膜白细胞介素(IL)-4,IL-9表达,蛋白质印迹法检测肠道蛋白酶激活受体2(PAR-2)表达,磁珠分选技术分离肠系膜淋巴结(MLN) DC和脾脏CD4+/CD8+T淋巴细胞,并予以体外共培养,检测DC促淋巴细胞分泌细胞因子的能力.组间比较采用t检验或方差分析.结果模型组大鼠回盲部黏膜肥大细胞数量多于对照组(2.73±0.21比1.13 ±0.10,P=0.000),PAR-2及IL-4、IL-9的表达均高于对照组[2.13±0.81比0.42±0.29、(7.2±1.2)比(3.3±1.0) pg/ml、(7.3±1.3)比(5.2±0.6) pg/ml,P=0.026、0.000、0.001],回盲部DC也多于对照组(8.91 ±2.88比6.34±0.94,P=0.045),体外MLN DC与CD4+T细胞共培养上清中IL-4水平高于对照组[(1.22±0.33)比(0.80±0.48) pg/ml,P=0.000].结论 IBS大鼠肠道DC数目增多,其能通过激活T淋巴细胞产生细胞因子IL-4,使肥大细胞处于高敏状态甚至脱颗粒,从而导致IBS内脏高敏感的产生.
목적 탐토대서장도수돌세포(DC)개도비대세포활화대장역격종합정(IBS)형성적작용궤제.방법 20지SD대서안수자표수궤분성대조조급모형조(각10지),모형조이결직장확장연합속박응격건립대서내장고민감모형,이복부수축반사(AWR)실험평고대서내장민감성,면역조직화학방법검측회맹부결장조직표면분자CD103표체,갑분알람염색법계수회맹부결장점막비대세포,매련면역흡부시험(E LISA)법검측장점막백세포개소(IL)-4,IL-9표체,단백질인적법검측장도단백매격활수체2(PAR-2)표체,자주분선기술분리장계막림파결(MLN) DC화비장CD4+/CD8+T림파세포,병여이체외공배양,검측DC촉림파세포분비세포인자적능력.조간비교채용t검험혹방차분석.결과 모형조대서회맹부점막비대세포수량다우대조조(2.73±0.21비1.13 ±0.10,P=0.000),PAR-2급IL-4、IL-9적표체균고우대조조[2.13±0.81비0.42±0.29、(7.2±1.2)비(3.3±1.0) pg/ml、(7.3±1.3)비(5.2±0.6) pg/ml,P=0.026、0.000、0.001],회맹부DC야다우대조조(8.91 ±2.88비6.34±0.94,P=0.045),체외MLN DC여CD4+T세포공배양상청중IL-4수평고우대조조[(1.22±0.33)비(0.80±0.48) pg/ml,P=0.000].결론 IBS대서장도DC수목증다,기능통과격활T림파세포산생세포인자IL-4,사비대세포처우고민상태심지탈과립,종이도치IBS내장고민감적산생.
Objective To explore the role of altered characteristics of intestinal dendritic cell(DC) in the induction of visceral hyperalgesia through the activation of mast cells in a rat model of irritable bowel syndrome (IBS).Methods A total of 20 Sprague-Dawley rats were divided into modeling and control groups (n =10 each).The IBS rat model was established by combining colorectal distention with restraint stress.Abdominal withdrawal reflex (AWR) was employed to evaluate visceral sensitivity.The surface marker of intestinal DC was analyzed by immunohistochemistry.Toluidine blue staining was used to determine the number of mast cells (MC).The expressions of interleukin (IL)-4 and IL-9 in colonic mucosa were measured by enzyme-linked immunosorbent assay (ELISA) and the level of proteinase-activated receptor-2 (PAR-2) was measured by Western blot.Mesenteric lymph node (MLN) DC and splenic CD4 +/CD8 + T cells were isolated and purified by magnetic label-based technique.Cytokine production of MLN DC co-culturing with CD4 + or CD8 + T cells was determined.Results The number of colonic MC in modeling group was more than that in control group ((2.73 ± 0.21) vs (1.13 ± 0.10),P =0.000).The expressions of PAR-2,IL-4 and IL-9 in colonic mucosa were all higher than those in control group (2.13 ± 0.81 vs 0.42 ±0.29,(7.2 ± 1.2) vs(3.3 ± 1.0) pg/ml,(7.3 ± 1.3) vs (5.2 ±0.6) pg/ml,P =0.026,0.000,0.001).Co-cultured MLN DC with CD4+ T cells showed a predominant IL-4 secretion in the modeling group ((1.22 ± 0.33) vs (0.80 ± 0.48) pg/ml,P =0.000).Conclusion Increased colonic DC may stimulate CD4 + T cells to secrete a high level of IL-4 to cause the degranulation of mast cells and the generation of visceral hypersensitivity in IBS rats.