不可预期应激刺激对大鼠消化道免疫和微生物菌群的影响
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1.中国中医科学院针灸研究所,北京 100700; 2.洛阳市第一中医院,河南 洛阳 471000

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Effects of chronic heterotypic intermittent stress stimulation on gastrointestinal immunity and microflora in rats
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1.Institute of Acupuncture and Moxibustion CACMS, Beijing 100700, China. 2. Luoyang No.1 Hospital of TCM, Luoyang 471000

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    摘要:

    目的 通过对 9 d 不可预期应激刺激干预后大鼠的观察,分析不可预期应激刺激对胃肠道免疫系统和微生物种群的影响,为慢性应激对胃肠道功能的研究提供参考。 方法 24 只 SD 大鼠随机分为空白组和慢性应 激模型组。模型组大鼠连续 9 d 接受不可预期应激刺激。造模完成后随机选择 5 只正常组和模型组大鼠,采集其粪便样本进行肠道微生物多样性分析。两组各 12 只大鼠中,随机选取 4 只进行灌流固定,取胃窦、结肠组织进行形态学观察;其余 8 只大鼠取血浆和肾上腺组织检测其中皮质酮的含量。 结果 (1)9 d 不可预期应激刺激后,模型组大鼠 9 d 体重增值与正常组大鼠相比明显减少(P= 0. 001),模型组大鼠肾上腺组织中皮质酮含量与正常组大鼠相比明显降低(P= 0. 006);模型组大鼠血浆中皮质酮含量与正常组大鼠相比增加(P= 0. 025)。 (2) 9 d 后模型 组大鼠的胃黏膜层上皮细胞排列较正常组稀疏,其厚度与正常组相比减少(P = 0. 034);模型组大鼠的胃黏膜固有 层中性粒细胞数量与正常组相比减少(P = 0. 016);模型组大鼠的结肠黏膜中中性粒细胞数量与正常组相比增多 (P= 0. 013)。 3)经过 9 d 慢性不可预期应激,Alpha 多样性分析显示,模型组大鼠粪便样本 OTU 中细菌种类与正常 组大鼠相比明显增加(P= 0. 001);其中模型组大鼠样本中脱硫弧菌科和螺杆菌科的占比为与正常组大鼠相比增加 (P= 0. 011,P= 0. 047);模型组大鼠样本中类杆菌科的占比与正常组大鼠相比明显减少,差异具有极显著的统计学意义(P= 0. 001)。 结论 9 d 不可预期应激刺激可以造成一个稳定的慢性应激大鼠模型,慢性应激所导致的免疫抑制与大鼠肠道微生物菌群失调伴随发生。

    Abstract:

    Objective To observe the effects of chronic heterotypic intermittent stress ( HIS ) on the gastrointestinal immune system, particularly in terms of (1) the morphology of the gastric and colonic mucosa, and (2) intestinal microbial diversity. Methods We randomly divided 24 SD rats into a control group and a model group. The rats in the model group received HIS for 9 days. After the 9 days of modeling, five rats in the control and model group were randomly selected, and fecal samples were collected for gut microbial diversity analysis. Four of the 12 rats from each of the two groups were randomly selected for perfusion and fixation, and tissues from the gastric antrum and colon were taken for morphological observation. Plasma and adrenal tissues were taken from the remaining eight rats for assessment of corticosterone content. Results (1) After 9 days of HIS, the rats in the model group had gained significantly less weight than those in the control group (P= 0. 001). Furthermore, the corticosterone content in adrenal tissue was significantly lower in the model group vs. the control group (P= 0. 006), and the corticosterone content in plasma was higher in the model group vs. the control group (P= 0. 025). (2) After HIS, the arrangement of gastric mucosal epithelial cells in the model group was thinner than that in the control group (P= 0. 034). Additionally, the number of neutrophils in the lamina propria of the gastric mucosa in the model group was lower than that in the control group (P= 0. 016). The number of neutrophils in the colonic mucosa in the model group was higher than that in the control group (P= 0. 013). (3) After 9 days of HIS, Alpha diversity analysis showed that the number of operational taxonomic units in the fecal samples from the model group was significantly higher than that in the control group (P= 0. 001). The proportion of desulfovibrionaceae and helicobacteraceae in the model group was higher than that in the control group (P= 0. 011, P= 0. 047). Furthermore, the proportion of bacteroidaceae in the model group was significantly lower than that in the control group ( P= 0. 001). Conclusions Nine days of HIS induced a stable rat model of chronic stress in which chronic stress-induced immunosuppression occurred concomitantly with dysregulation of the gut microbial flora.

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王晓宇,马富强,张晓宁,何 伟,宿杨帅,万红叶,胡 玲,景向红.不可预期应激刺激对大鼠消化道免疫和微生物菌群的影响[J].中国比较医学杂志,2020,30(8):1~9.

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  • 收稿日期:2019-10-09
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  • 在线发布日期: 2020-09-02
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