摘要
目的探讨生命早期益生菌鼠李糖乳杆菌GG(LGG)肠道定植对子代小鼠肠道屏障及肠道发育的影响及其可能的作用机制。方法选择SPF级6只6周龄野生型C57BL/6雌鼠随机分为实验组和对照组,实验组为LGG活菌组,对照组为LGG灭活菌组,分别予108 cfu/ml LGG活菌或LGG灭活菌灌胃直至自然分娩。两组子代小鼠分别于出生第1~5天继续给予107 cfu/ml LGG活菌或LGG灭活菌灌胃。记录3周子代小鼠体重变化;于第2、3周检测子代小鼠LGG菌的定植情况;运用Real-time PCR方法评价肠道促炎因子及紧密连接分子mRNA变化;运用HE、免疫组化、免疫荧光染色法及酶联免疫吸附法评价3周子鼠的肠道屏障情况。结果实验组的子代小鼠与对照组相比,第1周时体重相比无明显差异,第2周和第3周体重有所增加[第2周:(4.326±0.140)g比(3.790±0.240)g,t=3.707,P=0.006;第3周:(8.040±0.370)g比(7.295±0.326)g,t=3.130,P=0.011];仅在实验组子代小鼠的粪便中可检测出LGG定植。肠道定植可促进小肠绒毛长度及结肠隐窝深度明显增长[空肠:(320.000±22.514)μm比(265.100±15.611)μm,t=8.258,P<0.001;回肠(150.500±13.099)μm比(111.000±11.308)μm,t=9.958,P<0.001;结肠(295.000±15.209)μm比(233.100±6.678)μm,t=9.129,P<0.001]。与LGG灭活组相比,实验组子代小鼠结肠隐窝中杯状细胞的数量增加[(35.24 ±1.370)个比(11.62±0.780)个,t=15.000,P<0.001],促炎因子IFN-γ(0.512±0.206比1.280±0.232,t=4.970,P=0.001)、IL-6(0.941±0.215比1.364±0.271,t=2.452,P=0.040)、IL-10(0.744±0.294比1.341±0.320,t=2.762,P=0.025)和TNF-α(2.581±0.500比3.702±0.150,t=2.553,P=0.034)mRNA的相对表达水平降低,肠道组织紧密连接分子(Claudin3)(1.881±0.172比1.283±0.152,t=4.932,P=0.001)和闭锁蛋白分子(Occludin)(1.164±0.342比0.812±0.224,t=3.67,P=0.016)表达含量显著增加(P<0.01)。结论生命早期LGG定植可通过抑制肠道低度炎症进而保护肠道屏障。本研究将为生命早期补充益生菌进而防�
Objective To investigate the effects of Lactobacillus rhamnosus GG (LGG) colonization in early life on intestinal barrier and intestinal development in offspring mice and its possible mechanism. Methods Six C57BL/6 pregnant mice with the same conception time of 6 weeks were selected and randomly divided into experiment group given 108 cfu/ml LGG live bacteria and control group given LGG inactivated bacteria by gavage from the 18th day of pregnancy until natural birth. The progeny mice in the two groups were continued to be gavaged with 107 cfu/ml of LGG live bacteria or LGG inactivated bacteria on days 1-5 of birth. The body weight changes of 3 week'progeny mice were recorded. The colonization of LGG bacteria in offspring mice was detected at 2nd and 3rd weeks. The mRNA of intestinal proinflammatory cytokines and tight junction molecules were evaluated by real-time PCR method. HE, immunohistochemistry, immunofluorescence staining and enzyme-linked immunosorbent assay were used to evaluate the intestinal barrier of 3-week old off spring mice. Results Compared with the control group, the progeny mice of the experiment group showed no significant difference in body weight at the first week, and the body weight increased at the second week and the third week [2nd week:(3.790±0.240)g vs.(4.326±0.140)g, t=3.707, P=0.006;3rd week:(7.295±0.326)g vs.(8.040±0.370)g, t=3.130, P=0.011]. LGG colonization can be detected only in the feces of progeny mice in the experiment group. Intestinal colonization can promote the growth of small intestine villi and colon crypt depth[jejunum:(320.000±22.514)μm vs.(265.100±15.611)μm, t=8.258, P<0.001;ileum:(150.500±13.099)μm vs.(111.000±11.308)μm, t=9.958, P<0.001;colon:(295.000±15.209)μm vs.(233.100±6.678)μm, t=9.129, P<0.001]. Compared with the control group, the number of goblet cells in the colonic crypt of the experiment group increased(11.62±0.780 vs. 35.24 ±1.370, t=15.000, P<0.001), and the relative mRNA expression levels of pro-inflammatory factors as IFN-γ(1.280
作者
金歌
刘响
谢润祥
郭子宣
孙悦
刘天宇
王邦茂
曹海龙
Jin Ge;Liu Xiang;Xie Runxiang;Guo Zixuan;Sun Yue;Liu Tianyu;Wang Bangmao;Cao Hailong(Department of Gastroenterology and Hepatology, General Hospital of Tianjin Medical University, Tianjin 30052, China)
出处
《中华临床营养杂志》
CAS
CSCD
2019年第2期101-106,共6页
Chinese Journal of Clinical Nutrition
基金
国家自然科学基金资助项目(81741075).
关键词
生命早期
鼠李糖乳杆菌
低度炎症
肠道屏障
Early life
Lactobacillus rhamnosus GG
Low-grade inflammation
Intestinal barrier
