“五七”至“七七”阴虚体质女性肠道菌群及宿主NF-κB信号通路共变化机制研究

According to the statement about females’ physiological characteristics in Huang Di Nei Jing (Huangdi’s Canon of Medicine) , women’s degenerating period starts from 35 years to 49 years of age. Modern researches reveal that yin-deficiency constitution is closely related to senility, and the intestinal flora participate in the host’s aging process. Existence with the host gene is the general change feature. Previous studies have revealed gut microorganism imbalance in yin-deficiency constitution and abnormal expression of correlation factors of cell aging classical NF-κB excitation pathway. However, the regulatory mechanism of general change correlation between intestinal flora and gene in women’s degenerating period with yin-deficiency constitution has not been confirmed. For the above reason, we try to hypothesize there are feature functional flora related to aging in women with yin-deficiency constitution and the common change regulation features of the hosts’ NF-κB signaling pathway. In our experiment，the constitution assessment professional standard will be adopted and females aged from 35 to 49 will be screened for yin-deficiency or balanced constitutions. The MiSeq sequencing technique is going to do technical evaluation of the vital functional flora. PCR will be used to detect NF-κB signaling pathway of the mRNA expression level of NFKBIA、CCL4、BCL2A1,etc. Meanwhile, with multivariable statistical approaches we intend to know some of the general change mechanism of the intestinal flora and NF-κB signaling pathway in women’s degenerating period with yin-deficiency constitution. This project is of great significance for yin-deficiency constitution women anti-aging based on the intestinal flora and gene.

依据《黄帝内经》关于女性生理特点的描述,“五七”至“七七”之年为女性“渐衰期”。现代研究发现，阴虚质与衰老密切相关, 同时肠道菌群参与宿主衰老过程,并与宿主基因存在共变化特征。前期研究提示，阴虚质存在肠道微生物失衡，及与细胞衰老经典NF-kB活化途径相关因子mRNA表达异常。而阴虚质女性渐衰期肠道菌群是否参与基因共变化调控尚未证实。基于此，提出“阴虚质女性渐衰期存在与衰老相关特征功能菌群，且与宿主NF-κB通路存在共变化调控特征”这一假说。本项目拟采用体质判定行业标准，在35至49周岁女性中筛选阴虚质和平和质，运用MiSeq测序技术分析阴虚质肠道菌群特征；运用荧光定量PCR进一步检测NF-κB通路相关因子NFKBIA、CCL4、BCL2A1等mRNA表达水平；同时采用多变量统计方法揭示肠道菌群与NF-κB通路共变化调控机制。本项目对于开展基于肠道菌群、基因的阴虚质女性延缓衰老具有重要意义。

依据《黄帝内经》关于女性生理特点的描述,“五七”至“七七”为女性“渐衰期”。现代研究发现，阴虚质与衰老密切相关, 同时肠道菌群参与宿主衰老过程,并与宿主基因存在共变化特征。前期研究提示，阴虚质存在肠道微生物失衡，及与细胞衰老经典NF-kB活化途 径相关因子mRNA表达异常。基于此，提出“阴虚质女性渐衰期存在与衰老相关特征功能菌群，且与宿主NF-κB通路存在共 变化调控特征”这一假说。本项目采用体质判定行业标准，在35至49周岁女性中筛选阴虚质和平和质，运用MiSeq测序技术分析阴虚质肠道菌群特征;运用荧光定量PCR进一步检测NF-κB 通路相关因子NFKBIA、CCL4、BCL2A1等mRNA表达水平;同时采用多变量统计方法揭示肠道菌群 与NF-κB通路共变化调控机制。研究发现，阴虚质中NFKBIA、CCL4 mRNA表达显著上调， BCL2A1 mRNA显著下调，推测阴虚质中CCL4上调激发慢性炎症，NFKBIA代偿性升高，BCL2A1下调降低生存，正好激活NF-κB信号通路的衰老途径，从而导致炎性衰老。获取了阴虚质女性结构特征肠道菌群，功能菌群预测到老化（Aging）、癌症（Cancers）、传染病（Infectious diseases）、碳水化合物代谢（Carbohydrate metabolism）、氨基酸代谢（Amino acid metabolism）、表征不佳（Poorly characterized）、脂质代谢（Lipid metabolism）等相关功能，这些与验证了阴虚质女人存在与衰老相关的功能菌群。同时进行基因和肠道菌去的相关性分析，我们发现与TAK1,NFKBIA,CCL4,BCL2A1和IL8的相关性密切的肠道菌群微生物在组间存在相对丰度和差异贡献度等区别。本项目初步验证了假说。本项目对于开展基于肠道菌群、基因的阴虚质女性延缓衰老具有重要意义。