Dynamic Microbial Shifts and Functional Analysis of Saliva Microbial Communities with Caries Children
目的 观察龋病和无龋koko体育app 唾液菌群的动态变化，同时对二者口腔微生态在糖代谢及多种氨基酸合成转运过程中的功能差异进行分析。 方法 采用宏基因组学的方法分别对10例无龋koko体育app 和10例龋病koko体育app 唾液菌群进行组成及功能分析，6个月后，进一步利用PacBio SMRT测序技术，分析两组koko体育app 口腔菌群变化，探索疾病和健康状态下，口腔菌群变化规律，从组成到功能对koko体育app 口腔微生态进行全面解读。 结果 随时间推移，两组koko体育app 口腔微生态组成发生显著改变，在门水平上，菌群变化趋势一致，Firmicutes比例增加，而Actinobacteria和Bacteroidetes比例降低。在属水平上，两组koko体育app 唾液菌群的变化出现差异，其中，Lactobacillus、Methylobacterium和Megasphaera在龋病组koko体育app 中的丰度呈上升趋势，而在无龋组koko体育app 中呈下降趋势。在种水平上，隶属于乳杆菌属的L. fermentum、L. gasseri、L.oris、S. downei等菌株在龋病koko体育app 中呈上升趋势，但在无龋koko体育app 中始终处于较低水平。S. gordonii以及S. mutans在龋病koko体育app 中有一定程度下降，但在无龋koko体育app 中始终处于低水平。S. mutans和C. gracilis等菌种与龋失补牙数（decayed, missing and filled teeth, dmft）呈正相关，而N. flavescens与dmft呈负相关。三羧酸循环关键节点相关基因，gltA、icd以及mqo，谷氨酸生成相关基因gudB以及精氨酸生成相关基因argAB/C/J在无龋koko体育app 中丰度显著增加。同时，电子传递链中的NADH脱氢酶相关基因nuoB/C/D/E/H/I/J/K/L/M在无龋koko体育app 中的丰度显著增加。 结论 koko体育app 唾液菌群是动态变化的，其变化趋势与口腔健康状态有关，健康的口腔微生态中有更广泛的氧化磷酸化，谷氨酸以及精氨酸等氨基酸生成和转运也更为活跃。Abstract: Objective To observe the dynamic changes in the salivary microbiota of children with dental caries and those who were caries-free and to analyze the functional differences in the oral microecology of the two groups during the course of sugar metabolism and the synthesis and transport of multiple amino acids. Methods Ten children with dental caries and 10 caries-free children were enrolled. We employed Illumina metagenomics technology to analyze the composition and function of salivary microbiome in children with and without caries. Six months later, PacBio single-molecule long-read sequencing technology was used to analyze the changes over time in the oral microbial communities of the two groups. We studied the patterns of change in the oral microbial communities under diseased or healthy conditions and attempted to offer a comprehensive interpretation of children’s oral microbiota in terms of its composition and functions. Results The composition of the oral microbiota of children with or without dental caries changed significantly over time. At the phylum level, changing trends in the salivary microbial communities of children with dental caries were in line with those in caries-free children. In these microbial communities, increased proportions of Firmicutes and decreased proportions of Actinobacteria and Bacteroidetes were found in the two groups. At the genus level, however, the two groups showed significantly different changes of the salivary microbial communities. Upward trends in the abundance of Lactobacillus, Methylobacterium, and Megasphaera were found in the caries group, while the abundance of these genera in the caries-free group showed downward trends. At the species level, L. fermentum, L. gasseri, L. oris, S. downei, and some other species belonging to the genus Lactobacillus showed upward trends in the saliva of children with caries, while they consistently stayed at a relative low level of abundance in caries-free children. The abundance of S. gordonii and S. mutans decreased to a certain extent in children with dental caries, but the abundance of S. gordonii and S. mutans in caries-free children were always at a low level. Species such as S. mutans and C. gracilis were positively correlated to the sum of decayed, missing and filled teeth (dmft), while N. flavescens was negatively correlated to dmft. gltA, icd, and mqo, the key genes related to tricarboxylic acid (TCA) cycle, gudB, a glutamate synthesis-related gene, and argAB/C/J, arginine synthesis-related genes, were significantly increased in caries-free children. In addition, the abundance of the NADH dehydrogenase-related gene nuoB/C/D/E/H/I/J/K/L/M in the electron transport chain increased significantly in caries-free children. Conclusion Dynamic changes were found in the oral microbiota of children with or without caries. The trends of microbial shifts over time were associated with the oral health status. Oxidative phosphorylation and the synthesis and transport of amino acids such as glutamate and arginine in the oral microecology were more active in caries-free children.
图 4 糖代谢及氨基酸合成过程中关键基因差异分析
Figure 4. 🌌 Analysis of differences in key genes involved in glucose metabolism and amino acid synthesis
KEGG: Kyoto Encyclopedia of Genes and Genomes database. A: Key gene abundance for TCA cycle; B: Key gene abundance for glutamate and arginine cycle key genes synthesis; C: Abundance of NADH dehydrogenase related genes. *P<0.05. n=10.
图 5 腐胺及氨基酸运输过程关键基因差异分析
Figure 5. 🤡 Analysis of differences in key genes involved in putrescine and amino acid transportation
ESS: Electrochemical sodium symporter; NSS: Neurotransmitter sodium symporter; AGCS: Alanine or glycine cation symporter. A: Key gene abundance for putrescine transport system; B: The ESS family represents glutamate: Na+ symporter; The NSS family represents neurotransmitter: Na+ symporter; The AGCS family represents alanine or glycine: cation symporter. *P<0.05. n=10.
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