Supplementary Material for: The Trajectory of Successful Aging: Insights from Metagenome and Cytokine Profiling

Introduction: The longevity is influenced by of genetic, environmental, and lifestyle factors. The specific changes that occur in the gut microbiome during the aging process, and their relationship to longevity and immune function, have not yet been fully understood. The ongoing research other microbiome based to longevity cohort in Kazakhstan, provides preliminary information on longevity-related aging, where cytokine expression is associated with specific microbial communities and microbial functions. Methods: Metagenomic shotgun sequencing study of 40 long-lived individuals aged 90 years and over were carried out, who were conditionally healthy and active, able to served themselves, without a history of serious infection and cancer, who had not taken any antimicrobials, including probiotics. Blood serum weas analyzed to clinical and laboratory characteristics. The cytokine and chemokine profile in serum and stool samples was assessed using Multiplex analysis. Results: We found a significant increase in the expression of pro-inflammatory cytokines IL-1a, IL-6, 12p70, IP-10, IFN-α2, IL-15 TNFa, as well as chemokines MIP-1a/CCL3 and MIP-1b/CCL4, chemokine motif ligands MCP-3/CCL7 and MDC/CCL22(1c). Nonagenerians and centenarians demonstrated a greater diversity of core microbiota genera and showed an elevated prevalence of the genera Bacteroides, Clostridium, Escherichia and Alistipes. Conversely, there was a decrease in the abundance of the genera Ruminococcaceae, Fusicatenibacter, Dorea, as well as the species Fusicatenibacter saccharivorans. Furthermore, functional analysis revealed that the microbiome in long-lived group has a high capacity for lipid metabolism, amino acid degradation, and potential signs of chronic inflammatory status. Conclusion: Long-lived individuals exhibit an immune system imbalance and observed changes in the composition of the gut microbiota at the genus level between to the two age groups. Age-related changes in the gut microbiome, metabolic functions of the microbial community, and chronic inflammation all contribute to immunosenescence. In turn, the inflammatory state and microbial composition of the gut is related to nutritional status.

引言：长寿受遗传、环境与生活方式因素共同调控。衰老过程中肠道菌群发生的特异性变化，及其与长寿和免疫功能的关联，目前尚未完全阐明。本次在哈萨克斯坦开展的基于菌群的长寿队列研究，为长寿相关衰老机制提供了初步研究数据，该研究中细胞因子表达与特定微生物群落及微生物功能密切相关。方法：本研究针对40名90岁及以上的长寿个体开展宏基因组鸟枪测序（Metagenomic shotgun sequencing）分析。入选对象需满足以下条件：健康状况良好、生活可自理、无严重感染及癌症病史，且未服用过包括益生菌在内的任何抗菌药物。采集受试者血液血清，开展临床及实验室特征检测。采用多重分析（Multiplex analysis）技术，对血清及粪便样本中的细胞因子与趋化因子谱进行检测评估。结果：本研究发现促炎细胞因子IL-1α、IL-6、IL-12p70、IP-10、IFN-α2、IL-15及TNF-α的表达水平显著升高，同时趋化因子MIP-1α/CCL3、MIP-1β/CCL4、趋化因子基序配体MCP-3/CCL7及MDC/CCL22(1c)的表达亦显著上调。90岁以上长者与百岁老人的核心菌群属水平多样性更为丰富，拟杆菌属（Bacteroides）、梭菌属（Clostridium）、埃希氏菌属（Escherichia）及另枝菌属（Alistipes）的相对丰度显著升高。反之，瘤胃球菌科（Ruminococcaceae）、Fusicatenibacter属、多雷菌属（Dorea）的相对丰度则显著降低，同时Fusicatenibacter saccharivorans菌种的丰度也出现下降。此外，功能注释分析显示，长寿组人群的肠道菌群具备更强的脂质代谢、氨基酸降解能力，同时呈现出慢性炎症状态的潜在特征。结论：长寿人群存在免疫系统失衡现象，且两个年龄组的肠道菌群属水平组成存在显著差异。肠道菌群的增龄性变化、菌群群落的代谢功能异常及慢性炎症，共同促进了免疫衰老（immunosenescence）。反过来，肠道炎症状态与菌群组成亦与营养状况密切相关。