Th17细胞及其相关细胞因子在幽门螺杆菌感染中的作用研究

发布时间：2018-01-30 15:42

本文关键词： 幽门螺杆菌 Th17 白介素-23 白介素-17　出处：《南华大学》2011年硕士论文　论文类型：学位论文

【摘要】：目的:通过建立幽门螺杆菌(H. pylori)感染的C57BL/6小鼠模型,分析感染后不同时期小鼠胃组织IL-17、IL-23 mRNA及蛋白表达情况,以及脾脏单细胞悬液中Th17细胞应答情况,并分析感染后胃炎的严重程度与Th17-IL-17应答效应的相关性,同时设立治疗组检测上述指标的变化情况。为探讨IL-23/IL-17信号通路在H. pylori感染中的作用奠定基础,为H. pylori诱发的胃炎的免疫防御及治疗提供新的理论和实验依据。方法: (1)对6～8周龄的C57BL/6小鼠进行灌胃感染H. pylori,每隔48小时一次,共灌胃5次。在H. pylori感染后的4、8、12周分别处死小鼠,鉴定H. pylori感染小鼠模型是否成功。同时设立治疗组,采用三联疗法对灌胃感染4周后的小鼠进行治疗; (2)对对照组、感染组、治疗组小鼠分别取部分胃组织作H㤘E染色,显微镜下观察胃组织炎症变化; (3)采用Trizol法对胃组织及脾细胞提取RNA,RT-PCR检测细胞因子IL-17、IL-23 mRNA的表达情况; (4) ELISA检测胃组织匀浆上清中细胞因子IL-17、IL-23蛋白含量; (5)采用PMA+Ionomycin或H. pylori WCP对脾淋巴细胞进行刺激培养,流式细胞术检测脾脏单细胞悬液中Th17细胞应答情况。结果: (1)成功建立了H. pylori感染小鼠模型,感染组小鼠胃黏膜炎症程度随感染时间的延长而不断加重; (2)与对照组相比,H. pylori感染组小鼠的胃组织中IL-17、IL-23的表达在mRNA及蛋白水平均显著增高,H. pylori感染组小鼠的脾淋巴细胞中Th17细胞比例显著高于对照组小鼠的该比值; (3) H. pylori感染组小鼠IL-17、IL-23 mRNA及蛋白含量以及脾淋巴细胞中Th17细胞比例随感染时间延长而增加; (4)治疗组小鼠IL-17、IL-23表达量及脾淋巴细胞中Th17细胞比率,与治疗前即感染后4周的小鼠相比较均有所下降; (5)感染后不同时期小鼠胃黏膜的炎症程度与IL-17、IL-23的表达量存在正相关。结论: (1) H. pylori感染后IL-17、IL-23表达均上调; (2) H. pylori感染后可以诱导Th17细胞应答; (3) H. pylori感染后胃炎程度与胃组织IL-17、IL-23含量存在正相关。
[Abstract]:Objective: to establish a C57BL / 6 mouse model of Helicobacter pylori (H.pylori) infection and analyze the gastric tissue IL-17 of mice at different stages after infection. The expression of IL-23 mRNA and protein, and the response of Th17 cells in spleen single cell suspension. The relationship between the severity of post-infection gastritis and the response of Th17-IL-17 was analyzed. At the same time, a treatment group was set up to detect the changes of the above indexes, which laid a foundation for the study of the role of IL-23/IL-17 signaling pathway in H. pylori infection. To provide a new theoretical and experimental basis for the immune defense and treatment of H. pylori induced gastritis. Methods: (1) six weeks old C57BL / 6 mice were infused with H.pylorius every 48 hours, 5 times after H. pylori infection. After 12 weeks, the mice were killed to determine whether the model of H. pylori infection was successful or not. At the same time, the treatment group was set up, and the mice were treated with triple therapy after 4 weeks of gavage infection. (2) for the control group, the infection group and the treatment group, some gastric tissues were taken from the mice for H? The changes of inflammation in gastric tissue were observed under microscope. Trizol method was used to detect the expression of IL-17 IL-23 mRNA in gastric tissues and spleen cells by RT-PCR. (4) ELISA was used to detect the content of IL-17 and IL-23 in the supernatant of gastric tissue homogenate. 5) splenic lymphocytes were stimulated by PMA Ionomycin or H. pylori WCP. The response of Th17 cells in single cell suspension of spleen was detected by flow cytometry. Results: 1) the model of H. pylori infection in mice was successfully established, and the degree of gastric mucosal inflammation in infected mice was aggravated with the prolongation of infection time. (2) compared with the control group, the expression of IL-17 and IL-23 in the gastric tissues of mice infected with H. pylori was significantly increased in the level of mRNA and protein. The percentage of Th17 cells in spleen lymphocytes of pylori infected mice was significantly higher than that of control mice. The contents of IL-17 IL-23 mRNA and protein and the percentage of Th17 cells in spleen lymphocytes increased with the prolongation of infection time in mice infected with H. pylori. (4) the expression of IL-17 IL-23 and the ratio of Th17 cells in spleen lymphocytes in the treatment group were significantly lower than those in the control group (4 weeks after infection). (5) there was a positive correlation between the inflammatory degree of gastric mucosa and the expression of IL-17 and IL-23 in mice at different stages after infection. Conclusion: 1) the expression of IL-17 and IL-23 were up-regulated after infection with H. pylori. 2) Th17 cells could be induced by H. pylori infection. 3) the degree of gastritis after H. pylori infection was positively correlated with the content of IL-17 and IL-23 in gastric tissue.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R378

【相似文献】