目的 探讨当归多糖（Angelica Sinensis Polysaccharide，ASP）改善小鼠肠道微生态对脑缺血再灌注（Ischemia/reperfusion injury，I/R injury）的影响。 方法 将雄性SPF级昆明小鼠随机分为假手术组（Sham组），大脑中动脉栓塞组（MCAO组），当归多糖灌胃组（ASP组），粪菌移植组（Fecal microbiota transplantation，ASP-FMT），每组16只。连续给药7 d后建立MCAO模型，24 h后使用mNSS法检测小鼠神经功能缺陷评分; TTC染色法检测脑梗死相对面积；试剂盒法测定右侧大脑缺血半暗带组织SOD、MDA、GSH含量；Western blot测定炎症因子IL-1β、IL-5、IL-6、IL-10表达量；盲肠内容物16s rRNA测序分析肠道微生物多样性。 结果 与MCAO组相比，ASP干预后小鼠神经功能明显改善（P<0.001），脑梗死相对面积明显减少（P<0.01），氧化应激水平下降（P<0.05）；与MCAO组相比，ASP降低了脑组织IL-1β、IL-5及IL-6表达、上调IL-10表达量（P<0.05）；MACO后小鼠肠道菌群丰富度明显减少，益生菌比例下调，当归多糖灌胃或菌群移植均可部分恢复菌群丰富度，上调益生菌比例；肠道菌群的变化与脑组织损伤程度具有相关性。 结论 当归多糖通过改善肠道微生态，降低氧化应激水平，下调炎症因子水平，减轻脑缺血再灌注后神经功能损伤，减少梗死范围。
Abstract
Objective To investigate the modulatory effects of Angelica Sinensis Polysaccharide (ASP) on intestinal microbiota and its subsequent influence on cerebral ischemia/reperfusion (I/R) injury in murine models. Methods A cohort of 64 SPF level mice was stratified into four groups: sham operation group (Sham group), middle cerebral artery occlusion group (MCAO group), ASP gavage group (ASP group), and fecal microbiota transplantation group (ASP-FMT group), each comprising 16 subjects. Following a 7-day regimen of the respective interventions, the MCAO model was established. Neurological function deficits were quantified 24 hours post-MCAO using the modified Neurological Severity Scores (mNSS); cerebral infarct size was assessed via 2,3,5-triphenyltetrazolium chloride (TTC) staining; concentrations of superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione (GSH) in the ischemic penumbra were determined using assay kits. Additionally, the expression levels of inflammatory cytokines IL-1β, IL-5, IL-6, and IL-10 in brain tissues were quantified via Western blot, while 1 the diversity of intestinal microbiota was analyzed by 6s rRNA sequencing of cecal contents. Results Compared with the MCAO group, neurological function was markedly ameliorated after ASP intervention (P<0.001), cerebral infarct size was significantly reduced (P<0.01), and oxidative stress levels were attenuated (P<0.05). Compared with the MCAO group, the expression of pro-inflammatory cytokines IL-1β, IL-5, and IL-6 were substantially decreased by ASP intervention, while concurrently upregulated the anti-inflammatory cytokine IL-10 (P<0.05). Post-MCAO, a notable depletion in the richness of intestinal microbiota and a reduction in probiotic strains were observed. However, ASP administration or fecal microbiota transplantation partially restored microbiota richness and enhanced the proportion of probiotics. A correlation was established between the modulation of intestinal microbiota and the degree of brain tissue damage. Conclusions Angelica Sinensis Polysaccharide ameliorates intestinal microbiota, mitigates oxidative stress, modulates inflammatory cytokine profiles, and thereby attenuates neurological dysfunction and reduces infarct size following cerebral ischemia/reperfusion injury.
关键词
脑缺血再灌注损伤 /
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当归多糖 /
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肠道菌群 /
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炎症反应
Key words
Cerebral ischemia/reperfusion injury /
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Angelica Sinensis Polysaccharide /
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Intestinal microbiota /
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Inflammatory response
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基金
贵州省大学生创新创业计划训练项目(S202210660071)；贵州省卫健委科学技术基金项目（gzwkj2023-573）
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