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目的:探讨PI-IBS小鼠γδT细胞的表型和功能及其在PI-IBS发病中的作用。方法:旋毛虫感染小鼠,观察肠道炎症、腹壁撤退反射(内脏高敏感性)和结肠传输时间(肠道动力)。分别在感染后第2周和第8周处死动物,取末端回肠和近端结肠组织,免疫荧光组化,激光扫描共聚焦显微镜观察肠黏膜γδT细胞的分布和数量变化。收集PI-IBS小鼠肠道淋巴结和脾脏的淋巴细胞,单克隆抗体免疫磁珠分选法分离和纯化γδT细胞,3HTdR法检测其增殖情况,FACS检测其表面分子CD69、CD62L,ELISA检测细胞培养上清液中IFN-γ及IL-17的表达。结果:感染后第2周,PI-IBS小鼠肠道炎症明显,肠黏膜γδT细胞数量明显增加,明显增殖和活化,产生IL-17明显增加(P<0.01)。感染后第8周,PI-IBS小鼠肠道炎症基本消退,动物的腹壁撤退反射和结肠传输试验明显异常。肠黏膜γδT细胞数量仍然明显高于对照组小鼠,仍然有明显增殖、活化和产生IL-17(P<0.01)。结论:肠道γδT细胞增殖、活化及分泌IL-17可能参与PI-IBS的发病。
Objective: To investigate the phenotype and function of γδT cells in PI-IBS mice and its role in the pathogenesis of PI-IBS. Methods: Trichinella were infected with mice and intestinal inflammation, abdomen reflex (visceral hypersensitivity) and colon transit time (gut motility) were observed. The animals were sacrificed at the second week and the eighth week respectively. The distribution and number changes of γδT cells in the intestinal mucosa were observed by immunofluorescence staining and laser scanning confocal microscopy. The lymphocytes from the intestinal lymph nodes and spleen of PI-IBS mice were collected. The γδT cells were isolated and purified by the monoclonal antibody immunomagnetic beads sorting method. The proliferation of γδT cells was detected by 3HTdR method. The surface molecules CD69 and CD62L were detected by FACS. The supernatant IFN-γ and IL-17 expression. Results: At the second week after infection, intestinal inflammation of PI-IBS mice was obvious. The number of γδT cells in intestinal mucosa was significantly increased, and the proliferation and activation of PI-IBS mice significantly increased (P <0.01). At the 8th week after infection, the intestinal inflammation of PI-IBS mice subsided and the abdomen reflex and colonic transit test of animals were obviously abnormal. Intestinal mucosal γδT cell number is still significantly higher than the control group mice, still significant proliferation, activation and produce IL-17 (P <0.01). Conclusion: The proliferation, activation and secretion of IL-17 in γδT cells of gut may be involved in the pathogenesis of PI-IBS.