鼠李糖乳杆菌GG对后代肠道屏障的保护作用

Q4 Nursing

中华临床营养杂志 Pub Date : 2019-04-30 DOI:10.3760/CMA.J.ISSN.1674-635X.2019.02.007

Ge Jin, Xiang Liu, Runxiang Xie, Zixuan Guo, Yue Sun, Tianyu Liu, Bangmao Wang

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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. \n \n \nResults \nCompared 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. 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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±0.232 vs. 0.512±0.206, t=4.970, P=0.001), IL-6(1.364±0.271 vs. 0.941±0.215, t=2.452, P=0.040), IL-10(1.341±0.320 vs. 0.744±0.294, t=2.762, P=0.025) and TNF-α(3.702±0.150 vs. 2.581±0.500, t=2.553, P=0.034) in the experiment group decreased; the expression levels of the intimate tight junction molecules (Claudin3)(1.283±0.152 vs. 1.881±0.172, t=4.932, P=0.001)and the atresia protein molecule (Occludin)( 1.164±0.342 vs. 0.812±0.224, t=3.67, P=0.016) significantly increased. \n \n \nConclusion \nEarly life LGG colonization protects the intestinal barrier by inhibiting low-grade intestinal inflammation. 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引用次数: 0

摘要

目的探讨鼠李糖乳杆菌GG (Lactobacillus rhamnosus GG, LGG)早期定殖对子代小鼠肠道屏障和肠道发育的影响及其可能的机制。方法选择6周相同受胎时间的C57BL/6妊娠小鼠6只，从妊娠第18天起至自然分娩，随机分为试验组给予LGG活菌108 cfu/ml，对照组给予LGG灭活菌灌胃。两组子代小鼠在出生后第1-5天继续以107 cfu/ml的LGG活菌或LGG灭活菌灌胃。记录3周子代小鼠的体重变化。在第2周和第3周检测LGG细菌在子代小鼠中的定殖。实时荧光定量PCR法检测肠道促炎细胞因子和紧密连接分子mRNA表达水平。采用HE法、免疫组织化学法、免疫荧光染色法和酶联免疫吸附法对3周龄春小鼠肠道屏障进行评价。结果与对照组相比，实验组子代小鼠第1周体重差异无统计学意义，第2周和第3周体重增加[第2周:(3.790±0.240)g vs(4.326±0.140)g, t=3.707, P=0.006;第三周:g(7.295±0.326)和(8.040±0.370)g, t = 3.130, P = 0.011)。实验组仅在子代小鼠粪便中检测到LGG定殖。肠道定殖可促进小肠绒毛和结肠隐窝深度的生长[空肠:(320.000±22.514)μm vs(265.100±15.611)μm, t=8.258, P<0.001];回肠:μm(150.500±13.099)和(111.000±11.308)μm, t = 9.958, P < 0.001;结肠:μm(295.000±15.209)和(233.100±6.678)μm, t = 9.129, P < 0.001)。与对照组相比，实验组结肠隐窝杯状细胞数量增加(11.62±0.780 vs. 35.24±1.370,t=15.000, P<0.001)，促炎因子IFN-γ(1.280±0.232 vs. 0.512±0.206,t=4.970, P=0.001)、IL-6(1.364±0.271 vs. 0.941±0.215,t=2.452, P=0.040)、IL-10(1.341±0.320 vs. 0.744±0.294,t=2.762, P=0.025)、TNF-α(3.702±0.150 vs. 2.581±0.500,t=2.553, P=0.034) mRNA相对表达量降低;亲密紧密连接分子(Claudin3)(1.283±0.152比1.881±0.172,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)的表达水平显著升高。结论生命早期LGG定殖通过抑制低级别肠道炎症来保护肠道屏障。本研究将为生命早期补充益生菌以预防肠道疾病奠定实验基础。关键词:早期生活;鼠李糖乳杆菌GG;慢性炎症;肠屏障

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Protection effect of Lactobacillus rhamnosus GG supplied in early life on intestinal barrier in offspring

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±0.232 vs. 0.512±0.206, t=4.970, P=0.001), IL-6(1.364±0.271 vs. 0.941±0.215, t=2.452, P=0.040), IL-10(1.341±0.320 vs. 0.744±0.294, t=2.762, P=0.025) and TNF-α(3.702±0.150 vs. 2.581±0.500, t=2.553, P=0.034) in the experiment group decreased; the expression levels of the intimate tight junction molecules (Claudin3)(1.283±0.152 vs. 1.881±0.172, t=4.932, P=0.001)and the atresia protein molecule (Occludin)( 1.164±0.342 vs. 0.812±0.224, t=3.67, P=0.016) significantly increased. Conclusion Early life LGG colonization protects the intestinal barrier by inhibiting low-grade intestinal inflammation. This study will lay the experimental foundation for the supplementation of probiotics in early life so as to prevent intestinal diseases. Key words: Early life; Lactobacillus rhamnosus GG; Low-grade inflammation; Intestinal barrier

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来源期刊

中华临床营养杂志 Nursing-Nutrition and Dietetics

CiteScore

0.20

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2282

期刊介绍： The Chinese Journal of Clinical Nutrition was founded in 1993. It is the first professional academic journal (bimonthly) in my country co-sponsored by the Chinese Medical Association and the Chinese Academy of Medical Sciences to disseminate information on clinical nutrition support, nutrient metabolism, the impact of nutrition support on outcomes and "cost-effectiveness", as well as translational medicine and nutrition research. It is also a professional journal of the Chinese Medical Association's Parenteral and Enteral Nutrition Branch. The purpose of the Chinese Journal of Clinical Nutrition is to promote the rapid dissemination of knowledge on nutrient metabolism and the rational application of parenteral and enteral nutrition, focusing on the combination of multidisciplinary and multi-regional field investigations and clinical research. It mainly reports on nutritional risk screening related to the indications of parenteral and enteral nutrition support, "cost-effectiveness" research on nutritional drugs, consensus on clinical nutrition, guidelines, expert reviews, randomized controlled studies, cohort studies, glycoprotein and other nutrient metabolism research, systematic evaluation of clinical research, evidence-based case reports, special reviews, case reports and clinical experience exchanges, etc., and has a special column on new technologies related to the field of clinical nutrition and their clinical applications.