赵枫, 关素珍, 王凯, 付有娟, 刘红娅, 陈小惠, 戚发秋, 刘志宏. 枸杞多糖通过调节肠道菌群改善孕鼠慢性应激导致的子代情绪变化[J]. 环境与职业医学, 2020, 37(11): 1112-1118. DOI: 10.13213/j.cnki.jeom.2020.20185
引用本文: 赵枫, 关素珍, 王凯, 付有娟, 刘红娅, 陈小惠, 戚发秋, 刘志宏. 枸杞多糖通过调节肠道菌群改善孕鼠慢性应激导致的子代情绪变化[J]. 环境与职业医学, 2020, 37(11): 1112-1118. DOI: 10.13213/j.cnki.jeom.2020.20185
ZHAO Feng, GUAN Su-zhen, WANG Kai, FU You-juan, LIU Hong-ya, CHEN Xiao-hui, QI Fa-qiu, LIU Zhi-hong. Regulation of Lycium barbarum polysaccharide on gut microbiota and emotional change of offspring caused by chronic stress of pregnant rats[J]. Journal of Environmental and Occupational Medicine, 2020, 37(11): 1112-1118. DOI: 10.13213/j.cnki.jeom.2020.20185
Citation: ZHAO Feng, GUAN Su-zhen, WANG Kai, FU You-juan, LIU Hong-ya, CHEN Xiao-hui, QI Fa-qiu, LIU Zhi-hong. Regulation of Lycium barbarum polysaccharide on gut microbiota and emotional change of offspring caused by chronic stress of pregnant rats[J]. Journal of Environmental and Occupational Medicine, 2020, 37(11): 1112-1118. DOI: 10.13213/j.cnki.jeom.2020.20185

枸杞多糖通过调节肠道菌群改善孕鼠慢性应激导致的子代情绪变化

Regulation of Lycium barbarum polysaccharide on gut microbiota and emotional change of offspring caused by chronic stress of pregnant rats

  • 摘要: 背景

    肠道菌群通过脑-肠轴系统影响大脑功能及机体应激反应。肠道菌群和大脑之间的关系对于子代的生长发育尤为重要。

    目的

    了解孕鼠慢性应激对子代肠道菌群的影响,初步探索枸杞多糖(LBP)对因孕期慢性应激而导致的子鼠情绪改变的改善作用及其可能机制。

    方法

    选择成年SD大鼠,雌鼠24只,分为对照组、模型组和模型+LBP组,雄鼠20只,分为对照交配组和模型交配组,采用慢性不可预知温和应激建立孕期慢性应激动物模型,并用LBP灌胃干预。子鼠分为对照子鼠组、模型子鼠组和模型+LBP子鼠组。子鼠出生后第20天,收集新鲜粪便,采用Illumina MiSeq测序技术,测定子鼠粪便中微生物16S rRNAV3-V4区序列,并对群落结构和多样性进行交互分析;在出生后第28天称量体重并內眦静脉采血,采用放射免疫法测定血浆皮质酮水平;在出生后42天利用旷场实验、蔗糖偏好实验以及悬尾实验进行情绪变化的测定。

    结果

    模型组和模型+LBP组雌鼠在应激第7、14天血浆皮质酮水平分别较对照组升高(P < 0.05),提示雌鼠在孕期处于应激状态。LBP干预后,模型+LBP子鼠组体重(91.92±11.19)g高于模型子鼠组(81.06±15.71)g(P < 0.05);模型+LBP子鼠组水平运动(77.47±10.67)分高于模型子鼠组(67.32±11.39)分(P < 0.05);模型+LBP子鼠组1%蔗糖偏好百分比(67.00±6.00)%高于模型子鼠组(59.00±8.00)%(P < 0.05);模型+LBP子鼠组静止时间(113.39±16.75)s短于模型子鼠组(152.13±33.26)s(P < 0.05)。α多样性分析显示,模型+LBP子鼠组Sobs指数(377.20±30.03)高于模型子鼠组(311.00±11.87)(P < 0.05),模型+LBP子鼠组Ace指数(409.38±16.06)高于模型子鼠组(376.33±19.53)(P < 0.05);模型+LBP子鼠组厚壁菌门的相对丰度高于模型子鼠组,模型+LBP子鼠组毛螺菌科的相对丰度高于模型子鼠组。反应子鼠情绪的1%蔗糖偏好百分比与肠杆菌科丰度呈正相关(r=0.59),与拟杆菌科丰度呈正相关(r=0.44),与普雷沃氏菌科丰度呈负相关(r=-0.47);静止时间与肽链球菌科丰度呈负相关(r=-0.51);水平运动得分与双歧杆菌科丰度呈负相关(r=-0.48)(P < 0.05)。

    结论

    孕期慢性应激不仅导致母体处于较高的应激水平,而且其子代的应激水平也较高。LBP可以缓解孕期慢性应激导致的母体高应激状态,可能对子代肠道菌群的定植和物种的组成及多样性产生积极影响,改善因孕期慢性应激所导致的子鼠情绪变化。

     

    Abstract: Background

    Gut microbiota may affect brain function and stress response through the modulation of gut-brain axis (GBA). The relationship between gut microbiota and brain is particularly important for the growth and development of offspring.

    Objective

    This experiment investigates the effect of chronic stress of pregnant rats on the gut microbiota of progeny, and preliminarily explores the improving effect of Lycium barbarum polysaccharide (LBP) on the emotional change of offspring caused by chronic stress during pregnancy and its possible mechanism.

    Methods

    Twenty-four adult female SD rats were divided into a control group, a model group, and a model + LBP group; twenty adult male SD rats were divided into a control group and a model group. Chronic unpredictable mild stress (CUMS) was used to establish a rat model of chronic stress during pregnancy, and intragastric intervention was performed with LBP. The offspring rats were divided into a control offspring group, a model offspring group, and a model + LBP offspring group. The offspring's fresh stool samples were collected on postnatal day 20, the sequence of 16S rRNAV3-V4 regions of microorganisms in the feces of offspring was determined by Illumina MiSeq technique, and the interaction between microbial community structure and diversity were analyzed; their plasma corticosterone levels were detected by radioimmunoassay on postnatal day 28; their emotional changes were measured by open field test, sucrose preference test, and tail suspension test on postnatal day 42.

    Results

    The plasma corticosterone levels of the female rats in the model group and the model + LBP group on the 7th and 14th days of stress were higher than that of the control group (P < 0.05), suggesting that the female rats were in stress during pregnancy. After the LBP intervention, the model + LBP offspring group showed higher body weight, horizontal movement score, and 1% sucrose preference percentage, and shorter resting time than the model group did(91.92±11.19) g vs. (81.06±15.71) g; (77.47±10.67) vs. (67.32±11.39); (67.00±6.00)% vs. (59.00±8.00)%; (113.39±16.75)s vs. (152.13±33.26)s (P < 0.05). The results of α diversity analysis showed that the Sobs index of the model + LBP offspring group (377.20±30.03) was higher than that of the model offspring group (311.00±11.87) (P < 0.05), and the Ace index of the model + LBP offspring group (409.38±16.06) was higher than the model offspring group (376.33±19.53) (P < 0.05). The relative abundance of Firmicutes phylum in the model+LBP offspring group was higher than that in the model offspring group, so was Lachnospiraceae family. The offspring's 1% sucrose preference percentage was positively associated with Enterobacteriaceae abundance (r=0.59) and Bacteroidaceae abundance (r=0.44), and negatively associated with Prevotella Shan and Collins abundance (r=-0.47); resting time was negatively associated with Streptococcus abundance (r=-0.51); horizontal movement score was negatively associated with Bifidobacterium abundance (r=-0.48) (P < 0.05).

    Conclusion

    Chronic stress during pregnancy causes not only the mothers but also the offspring to be in a stress state. LBP can alleviate the maternal elevated stress level caused by chronic stress during pregnancy, positively affect the colonization of offspring's gut microbiota and the composition and diversity of microbial species, and improve the emotional changes of offspring caused by chronic stress during maternal pregnancy.

     

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