性激素变化对小鼠肠道菌群构成的影响
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北京林业大学生态与自然保护学院 北京 100091

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北京林业大学大学生创新创业训练计划项目(No. X202110022211),北京市自然科学基金项目(No. 5202016)


Effects of the Changes of Sex Hormones on Gut Microbiota Composition of Experimental Mice
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Beijing Forestry University

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    摘要:

    肠道菌群在维持宿主免疫和消化系统功能方面发挥着重要作用,肠道菌群的多样性和丰富度是衡量宿主健康状况的重要生理指标。性激素对动物生长发育发挥重要作用,但其对肠道菌群构成影响的相关实验研究相对较少。本研究以模式生物小鼠(Mus musculus)为对象,探究性激素的变化对小鼠肠道菌群构成的影响。采用外科手术方式建立小鼠去势模型,通过16S rRNA高通量测序技术,研究性激素对小鼠肠道菌群构成的影响。研究结果表明,雌性小鼠和雄性小鼠去势后,性激素水平显著下降。肠道菌群在门水平上,正常小鼠和去势小鼠肠道细菌群落均由拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)、埃普西隆杆菌门(Epsilonbacteraeota)、髌骨细菌门(Patescibacteria)、放线菌门(Actinobacteria)、软壁菌门(Tenericutes)、脱铁杆菌门(Deferribacteres)、酸杆菌门(Acidobacteria)和蓝藻细菌门(Cyanobacteria)组成,主要菌群为厚壁菌门和拟杆菌门物种,两门占物种相对丰度百分比达80%以上。Alpha多样性计算结果表明,雌性小鼠与雄性小鼠的正常组与去势对照组间的微生物区系均匀度无显著差异,Beta多样性结果进一步表明,正常组与去势组之间微生物区系结构无显著差异。由上述结果可以得到结论,性激素变化没有引起小鼠肠道菌群构成的差异。

    Abstract:

    [Objectives] Gut microbiota plays an important role in immune function and gastrointestinal system. The diversity and richness of gut microbiota are important physiological index to measure host health. Sex hormones play an important role on the growth and development of animals, but the effect of sex hormones on gut microbiota composition is not clear. [Methods] In this study, a mouse (Mus musculus) castration model was established by surgical method, fresh fecal samples of mice were continuously observed and collected during feeding, and the effects of sex hormones on the intestinal flora of mice were studied by 16S rRNA high-throughput sequencing technology. Usearch was applied to cluster reads with similarity above 97.0% and generate OTUs. Taxonomic annotations of feature sequences were processed by Bayesian classifier using SILVA as reference database. Statistics on composition in each sample were calculated at level of phylum, class, order, family, genus and species. QIIME was applied to obtain abundance of each species in samples, and distribution histogram at each taxonomic level were generated by certain R package. Alpha diversity metrics were evaluated by QIIME2. Beta diversity analysis was processed by QIIME. Blood was collected from the mice after they were fed for one month. The contents of estradiol and testosterone in serum of mice were determined by radioimmunoassay (RIA). [Results] Sex hormone levels in female and male mice decreased significantly after ovulation (Table 1). At phylum level, the intestinal bacterial communities of both normal mice and castrated mice were composed of Bacteroidetes, Firmicutes, Proteobacteria, Epsilonbacteraeota, Patescibacteria, Actinobacteria, Tenericutes, Deferribacteres, Acidobacteria and Cyanobacteria (Fig. 4). The populations of major bacteria found in normal and castrated mice were similar, with Firmicutes and Bacteroides as the dominant flora, and the sum of relative abundance percentages of the two flora was more than 80% (Table 4). Alpha diversity showed no significant difference in microflora evenness between the normal group and the castrated group (Table 5). Beta diversity showed no significant difference in microflora structure between the normal group and the castrated group. Followings are Beta diversity analysis based on four distance matrices: PCA analysis; PERMANOVA analysis and UPGMA analysis (Fig. 8). [Conclusion] The change of sex hormones did not cause the difference in the composition of the intestinal flora of mice.

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封暖暖,杨鹏程,赵浩汐,张晨曦,蒋元琳,韩香雨,胡德夫,刘树强.2023.性激素变化对小鼠肠道菌群构成的影响.动物学杂志,58(2):263-276.

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  • 收稿日期:2022-03-22
  • 最后修改日期:2023-02-08
  • 录用日期:2023-01-17
  • 在线发布日期: 2023-04-28
  • 出版日期: 2023-04-20