红景天苷对PM2.5暴露小鼠肠道微生物的调节作用

Moderating effect of salidroside on intestinal microbiota in mice exposed to PM2.5

  • 摘要:
    背景  红景天苷(SAL)对多器官系统具有保护作用。大气细颗粒物(PM2.5)暴露可造成肠道微生物紊乱,损害肠道健康。SAL对PM2.5暴露小鼠肠道微生物的调节作用有待深入研究。
    目的  探讨PM2.5对小鼠肠道菌群的影响及SAL的作用。
    方法  40只6~8周龄C57BL/6雄性小鼠随机分成4组,每组10只,即对照组、SAL组、PM2.5组、SAL+PM2.5组。SAL组、SAL+PM2.5组SAL灌胃(60 mg·kg−1),对照组、PM2.5组无菌生理盐水灌胃(10 mL·kg−1);PM2.5组、SAL+PM2.5组PM2.5悬液气管滴注(8 mg·kg−1),对照组、SAL组无菌生理盐水气管滴注(1.5 mL·kg−1)。2 d为1个实验周期,共进行10个周期,持续20 d。苏木素-伊红染色观察小鼠回肠组织病理变化。采集结肠内容物用于肠道微生物测序与短链脂肪酸(SCFAs)测定。
    结果  PM2.5组回肠组织炎症细胞浸润,SAL+PM2.5组仅有少量炎症细胞浸润。与对照组相比,PM2.5组Shannon降低(P<0.05),Simpson升高(P<0.05),该组肠道菌群多样性下降;SAL+PM2.5组Shannon较PM2.5组升高(P<0.05),Simpson降低(P<0.05),经SAL干预的小鼠肠道菌群多样性上升。主坐标分析结果显示PM2.5暴露组与对照组之间出现明显分离,对照组、SAL组与SAL+PM2.5组之间分离趋势不明显。非加权组平均法(UPGMA)聚类树结果显示对照组与SAL组最先被聚类在一起,两组再与SAL+PM2.5组聚类,最后三组与PM2.5组聚类。PCoA分析与UPGMA聚类结果均表示,PM2.5组均匀度与相似性显著降低。与对照组相比,PM2.5组拟杆菌门、Candidatus_Saccharimonas丰度降低(P<0.05),变形菌门、埃希氏菌属、拟杆菌属、普罗菲登斯菌属、肠球菌属与变形杆菌属丰度升高(P<0.05)。与PM2.5组相比,SAL+PM2.5组变形菌门、放线菌门、普罗菲登斯菌属、变形杆菌属丰度降低(P<0.05),Candidatus_Saccharimonas相对丰度显著升高(P<0.05)。PM2.5组丙酸、戊酸、己酸的含量较对照组降低(P<0.05),SAL+PM2.5组丙酸、异丁酸、丁酸、戊酸、己酸的含量较PM2.5组升高(P<0.05)。
    结论  呼吸道PM2.5暴露可引起小鼠肠组织病理改变、肠道微生物紊乱以及SCFAs的变化,而SAL能够在一定程度上起到保护作用。

     

    Abstract:
    Background Salidroside (SAL) has a protective effect on multiple organ systems. Exposure to fine particulate matter (PM2.5) in the atmosphere may lead to disruptions in gut microbiota and impact intestinal health. The regulatory effect of SAL on the gut microbiota of mice exposed to PM2.5 requires further investigation.
    Objective To evaluate gut microbiota disruption in mice after being exposed to PM2.5 and the potential effect of SAL.
    Methods Forty male C57BL/6 mice, aged 6 to 8 weeks, were randomly divided into four groups: a control group, an SAL group, a PM2.5 group, and an SAL+PM2.5 group, each containing 10 mice. In the SAL group and the SAL+PM2.5 group, the mice were administered SAL (60 mg·kg−1) by gavage, while in the control group and the PM2.5 group, sterile saline (10 mL·kg−1) was administered by gavage. In the PM2.5 group and the SAL+PM2.5 group, PM2.5 suspension (8 mg·kg−1) was intratracheally instilled, and in the control group and SAL group, sterile saline (1.5 mL·kg−1) was intratracheally administered. Each experiment cycle spanned 2 d, with a total of 10 cycles conducted over 20 d. Histopathological changes in the ileum tissue of the mice were observed after HE staining. Colon contents were collected for gut microbiota sequencing and short-chain fatty acids (SCFAs) measurements.
    Results The PM2.5 group showed infiltration of inflammatory cells in the ileum tissue, while the SAL+PM2.5 group exhibited only a small amount of inflammatory cell infiltration. Compared to the control group, the PM2.5 group showed decreased Shannon index (P<0.05) and increased Simpson index (P<0.05), indicating that the diversity of gut microbiota in this group was decreased; the SAL+PM2.5 group showed increased Shannon index compared to the PM2.5 group (P<0.05) and decreased Simpson index (P<0.05), indicating that the diversity of gut microbiota in mice intervened with SAL was increased. The principal coordinates analysis (PCoA) revealed a significant separation between the PM2.5 group and the control group, while the separation trend was less evident among the control group, the SAL group, and the SAL+PM2.5 group. The unweighted pair-group method with arithmetic means (UPGMA) clustering tree results showed that the control group and the SAL group clustered together first, followed by clustering with the SAL+PM2.5 group, and finally, the three groups clustered with the PM2.5 group. The PCoA and UPGMA clustering results indicated that the uniformity and similarity of the microbiota in the PM2.5 group were significantly decreased. Compared to the control group, the PM2.5 group showed decreased abundance of phylum Bacteroidetes and Candidatus_Saccharimonas (P<0.05) and increased abundance of phylum Proteobacteria, genus Escherichia, genus Bacteroides, genus Prevotella, genus Enterococcus, and genus Proteus (P<0.05). Compared to the PM2.5 group, the SAL+PM2.5 group showed decreased abundance of phylum Proteobacteria, phylum Actinobacteria, genus Prevotella, and genus Proteus (P<0.05), and increased abundance of Candidatus_Saccharimonas (P<0.05). The PM2.5 group showed reduced levels of propionic acid, valeric acid, and hexanoic acid compared to the control group (P<0.05), while the SAL+PM2.5 group showed increased levels of propionic acid, isobutyric acid, butyric acid, valeric acid, and hexanoic acid compared to the PM2.5 group (P<0.05).
    Conclusion Exposure to PM2.5 can cause pathological alterations, microbial dysbiosis, and disturbing production of SCFAs in intestinal tissue in mice. However, SAL can provide a certain degree of protective effect against these changes.

     

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