Analysis of intestinal flora in Mongolian female students under anxiety in a medical university
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摘要:
目的 探讨蒙古族学生焦虑情绪下肠道菌群变化, 为探究体内菌群与分泌物表达之间的关系提供依据。 方法 通过焦虑自评量表及半结构化访谈对某医学高校454名大学生焦虑情况展开调查, ≥50分者为焦虑情绪组、 < 50分者为对照组(无焦虑情绪组), 收取符合纳入标准且同意取样的被试粪便样本, 从两组中选取蒙古族女生提供的样本。收集每份粪便样品的细菌总DNA进行PCR扩增, 针对16S rDNA V3~V4区进行NovaSeq 2x250bp高通量测序, 并进行组间OTU分析、样品多样性分析、样品组间差异分析及LEfSe分析。 结果 蒙古族医学生焦虑检出率为23.86%, 汉族学生焦虑检出率为16.96%, 差异无统计学意义(P=0.07);组间Alpha多样性指数分析显示, Shannon、Simpson、Chao1、Observed species指数在女生焦虑情绪组与对照组组间差异无统计学意义(t值分别为8.0, 9.0, 6.0, 6.5, P值均>0.05)。在纲、目, 科, 属、种水平上, 焦虑情绪组与对照组在某些物种上差异有统计学意义(t=-2.26~2.57, -5.08~3.58, -2.65~2.09, P值均 < 0.05)。 结论 Alpha多样性指数显示焦虑情绪组与对照组肠道群落丰富度和多样性无差异, 但两组在不同分类水平上均存在差异物种, 提示焦虑情绪状态下肠道菌群结构会改变, 肠道菌群与焦虑情绪存在一定关联。 Abstract:Objective To investigate the change in intestinal flora in Mongolian students with anxiety, so as to provide basis for exploring the relationship between flora and secretion expression in vivo. Methods The Self-rating Anxiety Scale(SAS)was used to assess anxiety in medical college students; then a semi-structured interview was administered. Fecal samples that met the inclusion criteria were collected and divided into anxiety (SAS score≥50) and control groups (no anxiety, SAS score < 50) according to the standard score of SAS. Samples provided by Mongolian female students were selected from each group. The total bacterial DNA was extracted from the fecal samples for PCR amplification and NovaSeq 2x250bp high-throughput sequencing was performed for the V3-V4 region of 16S rDNA gene to obtain the biological information of the intestinal flora. The intergroup OTU, structural diversity, significant difference, and LEfSe analyses were performed with information mining of the literature think tanks. Results Anxiety existed in 23.86% of the Mongolian students, and 16.96% of the Han people. A Chi-square test showed no significant difference in detection of anxiety between Mongolian and Han college students (P=0.07). Analysis of the alpha diversity index showed that the Shannon index, Simpson index, Chao1 index, and Observed species did not differed significantly between the two groups(t=8.0, 9.0, 6.0, 6.5). The difference in abundance of some bacteria was significant at the Class, Order, Family, and Genus levels between the two groups(t=-2.26-2.57, -5.08-3.58, -2.65-2.09, P < 0.05). Conclusion The alpha diversity index showed that there was no significant difference in the abundance and diversity of intestinal flora between the two groups. While there were significant differences at different classification levels, the results suggest that the structure of intestinal flora can change in students with anxiety. -
Key words:
- Anxiety /
- Gastroin testinal tract /
- Bacteria /
- Students /
- Minority groups
1) 利益冲突声明 所有作者声明无利益冲突。 -
表 1 两组蒙古族女生肠道菌群Alpha多样性指数比较(x±s)
Table 1. Comparison of alpha diversity index between the two groups of Mongolian girls(x±s)
组别 人数 Chao1 Observed species PD whole tree Shannon Simpson Goods coverage 焦虑情绪组 5 217.87±22.42 204.00±27.46 13.83±1.55 4.75±0.38 0.91±0.03 1.00±0.00 对照组 5 237.53±26.52 221.00±23.84 14.82±1.71 5.00±0.34 0.92±0.03 1.00±0.00 t值 6.0 6.5 6.0 8.0 9.0 18.5 P值 0.22 0.25 0.22 0.42 0.55 0.25 
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表 2 两组蒙古族女生肠道细菌纲和目水平比较(x±s)
Table 2. Comparison between the 2 groups of Mongolian girls gut microbiota at the class and order level(x±s)
组别 人数 Negativicutes Bacilli Deltaproteobacteria Selenomonadales Lactobacillales Desulfovibrionales 焦虑情绪组 5 9.66×102±5.94×10-3 1.86×10-2±1.90×10-4 3.45×10-4±2.13×10-7 9.66×10-2±5.94×10-3 1.81×10-2±1.80×10-4 3.45×10-4±2.13×10-7 对照组 5 2.43x10-2±1.69×10-4 2.91x10-3±1.24×10-6 2.25x10-3±3.36×10-6 2.47×10-2±1.69x10-4 2.67×10-3±1.19×10-6 2.25×10-3±3.36×10-6 t值 2.06 2.54 -2.26 2.06 2.57 -2.26 P值 0.04 0.02 0.03 0.03 0.01 0.02
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表 3 两组蒙古族女生肠道细菌科水平比较(x±s)
Table 3. Comparison between the 2 groups of Mongolian girls gut microbiota at the family level(x±s)
组别 人数 Bacteroidaceae Veillonellaceae Rikenellaceae Acidaminococcaceae Tannerellaceae Peptostreptococcaceae 焦虑情绪组 5 1.16×10-1±1.02×10-2 9.41×10-2±6.42×10-3 3.12×10-3±1.74×10-5 2.69×10-3±3.28×10-5 4.43×10-3±3.96×10-5 1.34×10-2±5.83×10-5 对照组 5 2.43×10-1±1.08×10-2 7.34×10-3±2.0×10-4 2.13×10-2±2.84×10-4 1.73×10-2±2.43×10-4 1.19×10-2±3.09×10-5 9.69×10-4±1.65×10-6 t值 -1.96 2.38 -2.35 -1.97 -2.00 3.58 P值 0.04 0.02 0.02 0.04 0.04 0.00 组别 人数 Streptococcaceae Barnesiellaceae Marinifilaceae Desulfovibrionaceae FamilyXIII 焦虑情绪组 5 1.25×10-2±1.29×10-4 4.68×10-4±7.28×10-7 5.68×10-4±3.14×10-7 3.47×10-4±2.15×10-7 1.67×10-4±4.38×10-8 对照组 5 1.82×10-3±2.60×10-7 6.37×10-3±3.69×10-5 2.85×10-3±6.93×10-7 2.25×10-3±3.36×10-6 5.79×10-4±1.70×10-7 t值 2.10 -2.15 -5.08 -2.25 -1.99 P值 0.03 0.03 0.00 0.02 0.04
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表 4 两组蒙古族女生肠道细菌属和种水平比较(x±s)
Table 4. Comparison between the 2 groups of Mongolian girls gut microbiota at the genus and species level(x±s)
组别 人数 Bacteroides Alistipes Phascolarctobacterium Parabacteroides RuminococcaceaeUCG-013 焦虑情绪组 5 1.17×10-1±1.02×10-2 3.23×10-3±1.80×10-5 2.69×10-3±3.30×10-5 4.51×10-3±3.96×10-5 2.57×10-3±6.01×10-6 对照组 5 2.43×10 -1±1.08×10-2 2.14×10-2±2.85×10-4 1.74×10-2±2.44×10-5 1.19×10-2±3.11×10-5 7.76×10-3±1.89×10-5 t值 -1.95 -2.33 -1.97 -1.98 -2.32 P值 0.04 0.02 0.02 0.04 0.02 组别 人数 Barnesiella Odoribacter Bilophila Veillonella Streptococcus 焦虑情绪组 5 3.97×10-4±7.88×10-7 4.92×10-4±2.32×10-7 3.47×10-4±2.15×10-7 2.09×10-2±4.14×10-4 1.25×10-2±1.24×10-5 对照组 5 5.67×10-3±3.41×10-5 1.98×10-3±2.47×10-6 1.52×10-3±7.73×10-7 7.82×10-4±2.99×10-7 1.73×10-3±2.07×10-7 t值 -2.00 -2.03 -2.65 2.21 2.16 P值 0.04 0.04 0.01 0.03 0.03 组别 人数 Tyzzerella3 Streptococcus salivarius subsp. thermophilus Bacteroides sp.HPS0048 Actinomyces sp.oral strain Hal-1065 焦虑情绪组 5 1.45×10-3±1.53×10-6 7.48×10-3±6.71×10-3 5.82×10-5±1.30×10-4 2.12×10-4±2.21×10-4 对照组 5 2.12×10-4±2.25×10-7 4.91×10-4±3.10×10-4 2.68×10-4±1.86×10-4 5.58×10-6±1.25×10-5 t值 2.09 0.36 0.36 0.36 P值 0.03 0.01 0.03 0.03
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表 5 组间蒙古族女生核心菌群表
Table 5. The core flora of two groups of Mongolian girls
组别 物种信息 平均丰度的log10值 LDA值 P值 对照组 Marinifilaceae 3.45 3.15 0.01 (n=5) Rikenellaceae 4.33 4.01 0.03 Alistipes 4.33 4.01 0.03 Barnesiellaceae 3.80 3.41 0.01 Barnesiella 3.75 3.45 0.01 Acidaminococcaceae 4.24 3.85 0.03 Phascolarctobacterium 4.24 3.85 0.03 RuminococcaceaeUCG_013 3.89 3.44 0.04 焦虑情绪组 Negativicutes 4.98 4.56 0.03 (n=5) Veillonellaceae 4.97 4.64 0.02 Veillonella 4.32 4.04 0.00 Megasphaera 4.08 3.79 0.01 Selenomonadales 4.98 4.56 0.03 Pasteurellales 4.01 3.58 0.01 Pasteurellaceae 4.01 3.58 0.01 Haemophilus 3.97 3.56 0.01 Prevotella2 1.70 3.26 0.03
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