益生菌对肠易激综合征小鼠肠道黏膜CRF-R1表达及肠道屏障功能的影响  

The effect of Bifidobacterium infantis on intestinal CRF-R1 and intestinal barrier function in murine model with irritable bowel syndrome

作　　者：刘思濛 于泳[1,2] 黄煌[1,2] 丁一芮 白利梅[1,2] 李付广[3] 郑鹏远[1,2] 

机构地区：[1]郑州大学第二附属医院消化内科 [2]郑州大学医学微生态及临床营养研究所,河南郑州450014 [3]郑州大学基础医学院微生物学与免疫学教研室,河南郑州450001

出　　处：《中国微生态学杂志》2014年第9期993-997,1001,共6页Chinese Journal of Microecology

基　　金：国家自然科学基金(81370494);国家973计划前期研究专项(2011CB512006);河南省科技厅重点攻关(102102310400);郑州市创新型科技人才队伍建设工程基金(131PLJRC656);河南省卫生厅普通项目(201303086)

摘　　要：目的通过观察肠易激综合征（IBS）小鼠模型结肠黏膜肥大细胞上CRF-R1及结肠黏膜PAR-2、Claudin1~4等的表达变化,探讨IBS中应激通过肥大细胞引起肠道屏障功能障碍的可能机制并观察婴儿双歧杆菌的治疗作用。方法 30只雄性Balb/c小鼠随机分为对照组、模型组及婴儿双歧杆菌组。以束缚应激法建立IBS小鼠模型。婴儿双歧杆菌组给予婴儿双歧杆菌灌胃,而对照组及模型组给予等体积生理盐水灌胃。观测腹肌收缩反射（AWR）后处死小鼠。ELISA检测外周血中类胰蛋白酶的表达变化。免疫组织化学分析结肠黏膜CRF、PAR-2、Claudin1、Claudin2、Claudin3、Claudin4的表达情况。免疫荧光双标分析结肠黏膜CRF-R1在肥大细胞的表达情况。RT-PCR检测结肠CRF-R1mRNA的表达情况。结果与对照组相比,模型组小鼠外周血类胰蛋白酶表达量增加;结肠黏膜中CRF、PAR-2、Claudin2表达量、CRF-R1＋肥大细胞数目及CRF-R1 mRNA表达量增加,结肠黏膜Claudin1、Claudin3、Claudin4表达量降低,差异均有统计学意义（P〈0.05）。婴儿双歧杆菌干预后,该组小鼠外周血中类胰蛋白酶表达量降低;结肠黏膜中CRF、PAR-2、Claudin2表达量、CRF-R1＋肥大细胞数目及CRF-R1 mRNA表达量降低,结肠黏膜Claudin1、Claudin3、Claudin4表达量增高,差异均有统计学意义（P〈0.05）。结论 IBS小鼠模型中,婴儿双歧杆菌可以减轻肠道屏障功能障碍,其机制可能与抑制结肠黏膜肥大细胞上CRF-R1的表达而抑制肥大细胞的激活及其免疫因子的释放,从而降低结肠黏膜PAR-2的表达有关。Objective To investigate the effect of Bifidobacterium infantis on the expression of intestinal CRF-R1 in mast cells and the intestinal barrier function in murine model with irritable bowel syndrome（ IBS）. Methods Thirty Balb /c male mice were randomly divided into control group,model group and Bifidobacterium infantis group.The IBS mice models were established by chronic restraint stress. The Bifidobacterium infantis group received intragastrical Bifidobacterium infantis daily for 14 days,while the control and model groups were treated with equal volume of saline. The mice were killed at the day 14. The weight and the abdominal withdrawal reflex（ AWR） of mice were assessed. The level of tryptase in blood serum was detected by ELISA. The expression of CRF,PAR-2,Claudin1,Claudin2,Claudin3 and Claudin4 in colon mucosa was analyzed by immunohistochemistry. The expression of CRF-R1 in mast cells in colon mucosa was detected by double immunofluorescence staining. The expression of CRF-R1 mRNA in colon was detected by reverse transcription polymerase chain reaction（ RT-PCR）. Results Compared to the control group,the level of tryptase in peripheral blood,the expression of CRF,PAR-2,Claudin2,CRF-R1＋mast cells,and CRF-R1 mRNA in colon increased significantly,while the expression of Claudin1,Claudin3 and Claudin4 decreased significantly in model group（ P〈 0. 05）. After the treatment of Bifidobacterium infantis,the level of tryptase in peripheral blood,the expression of CRF,PAR-2,Claudin2,CRF-R1＋mast cells,and CRF-R1 mRNA in colon decreased significantly,while the expression of Claudin1,Claudin3 and Claudin4 increased significantly as compared with the model group（ P〈 0. 05）. Conclusion Bifidobacterium infantis can reduce the expression of intestinal CRF-R1 in mast cells and alleviate the intestinal barrier dysfunction in murine model with irritable bowel syndrome.

关 键 词：肠易激综合征 益生菌 肥大细胞 肠道屏障功能 

分 类 号：R378.991[医药卫生—病原生物学]