基于16S rRNA测序分析富锶水对老龄小鼠肠道菌群的影响

Effects of Sr-rich water on gut microbiota in aged mice based on 16S rRNA sequencing

目的探讨富锶水长期干预对老龄小鼠肠道菌群的影响。方法取16只18月龄雄性C57BL/6小鼠,采用锶含量为6.11 mg/L的复合型矿泉水连续干预8周后,收集小鼠新鲜粪便进行16S rRNA测序,分析富锶水对老龄小鼠肠道菌群的影响,并通过KEGG分析富锶水对老龄小鼠代谢通路富集的影响。结果富锶水干预后与对照组相比,老龄小鼠肠道菌群结构发生变化。聚类分析发现,两组共有595个相同OTUs,对照组特有OTUs为1108个,富锶水组特有OTUs为44个。富锶水组的菌群多样性发生改变:与对照组相比,富锶水组老龄小鼠肠道疣微菌门(t=−2.643,P=0.033)、放线菌门(t=−3.389,P=0.006)、梭杆菌门(t=−3.531,P=0.004)、疣微菌纲(t=−2.643,P=0.033)、疣微菌目(t=−2.651,P=0.019)、阿克曼菌科(t=−2.652,P=0.033)、阿克曼菌属(t=−2.652,P=0.033)、嗜黏蛋白阿克曼菌(t=−2.652,P=0.033)相对丰度显著增加,脱铁杆菌门(t=2.337,P=0.047)、α−变形杆菌纲(t=2.506,P=0.041)、脱硫弧菌纲(t=2.337,P=0.047)、Marinifilaceae(t=2.438,P=0.032)、脱铁杆菌科(t=2.337,P=0.047)、Lachnospiraceae_NK4A136_group(t=2.508,P=0.039)相对丰度显著减少。KEGG富集分析发现,富锶水干预后氨基酸相关酶、精氨酸和脯氨酸代谢、酪氨酸代谢、糖基转移酶、磷酸肌醇代谢、磷脂酰肌醇信号系统等代谢通路富集程度显著高于对照组(P<0.05)。结论长期饮用富锶水对老龄小鼠肠道菌群的结构产生影响,不同丰度的肠道菌群可能通过调节细胞结构、能量代谢、肌肉收缩等途径来达到改善肠道结构,维持健康的肠道屏障,预防多种疾病发生的目的,但具体作用机制有待深入研究。

Objective To explore the effects of strontium-rich water on the intestinal flora in aged mice.Methods Sixteen 18-month-old male C57BL/6 mice were treated with fresh compound mineral water containing 6.11 mg/L strontium for 8 weeks.Fresh feces were collected for 16S rRNA sequencing and bioinformatics analysis.The enrichment of functional genes in metabolic pathways was analyzed based on KEGG database.Results Compared with the control group,the abundance and diversity of gut microbiota in the aged mice were significantly different after strontium-rich water intervention.The cluster analysis showed that there were 595 common species in the control group and the Sr-rich water(SRW)group at the OTU level,1108 unique bacterial species in the control group and 44 unique species in the Sr-rich water group.The diversity of bacteria in SRW group was changed.Compared with the control group,the relative abundances of Verrucomicrobiota(t=−3.389,P=0.006),Actinobacteriota(t=−3.389,P=0.006),Fusobacteriota(t=−3.531,P=0.004),Verrucomicrobiae(t=−3.531,P=0.004),Verrucomicrobiales(t=−2.651,P=0.019),Akkermansiaceae(t=−2.652,P=0.033),Akkermansia(t=−2.652,P=0.033)and Akkermansia muciphila(t=−2.652,P=0.033)significantly increased,while those of Deferribacteres(t=2.337,P=0.047),Alphaproteobacteria(t=2.506,P=0.041),Deferribacteres(t=2.337,P=0.047),Marinifilaceae(t=2.438,P=0.032),Deferribacteraceae(t=2.337,P=0.047)and Lachnospiraceae_NK4A136_group(t=2.508,P=0.039)significantly reduced.KEGG analysis revealed that the metabolic pathways were mainly enriched in amino acid related enzymes,arginine and proline metabolisms,tyrosine metabolism,glycosyltransferase,phosphatidylinositol signaling system,and phosphatidylinositol metabolism.Conclusion Long-term consumption of strontium-rich water can affect the structure of intestinal flora in aged mice.Intestinal flora with different abundances may improve intestinal structure,maintain healthy intestinal barrier and prevent various diseases by regulating cell structure,energy metabolism and muscle contraction,etc.However,the specific mechanism of action needs be further studied.