基于PCR-DGGE技术分析中老年人肠道菌群结构与免疫功能的关系

发布时间：2018-08-09 17:05

【摘要】：目的通过检测中老年人的各项免疫指标和肠道菌群组成,分析中老年人免疫功能与其肠道菌群结构的关系,探索高龄引起的肠道菌群结构变化对中老年人免疫功能可能的影响。方法选取50~75周岁,健康中老年人99人作为研究对象,抽取空腹静脉血,检测CD3~+T细胞数、CD4~+T细胞数、CD8~+T细胞数及血清中的IgA、IgM、IgG。收集各研究对象的粪便,提取粪便细菌基因组DNA,采用细菌16S rRNA V6~V8区通用引物进行PCR扩增,对扩增产物进行变性梯度凝胶电泳。采用聚类分析、主成分分析、方差分析等解析研究对象肠道菌群结构特征及与其免疫指标的相关性。结果研究对象肠道菌群组成表现出高度多态性和显著的个体差异,CD3~+T细胞和CD4~+/CD8~+水平与肠道菌群结构有相关性,CD4~+/CD8~+偏高的人群与CD4~+/CD8~+偏低和正常人群相比肠道菌群丰富度(S)偏低,但是未观察到IgA、IgM、IgG与肠道菌群结构存在明显相关。结论中老年人肠道菌群结构与免疫水平相关,肠道菌群多样性的减少可能会影响其机体CD8~+T细胞为中心的非特异性自然免疫功能。
[Abstract]:Objective to analyze the relationship between immune function and intestinal flora structure in middle-aged and elderly people by detecting the immune indexes and composition of intestinal flora, and to explore the possible effects of the changes of intestinal flora structure on immune function in middle-aged and elderly people. Methods 99 healthy adults aged 50 to 75 years were selected as subjects. Fasting venous blood samples were collected to detect the number of CD3 ~ T cells and the number of CD8 ~ T cells in serum. The feces of all the subjects were collected, and the genomic DNA of fecal bacteria was extracted. The amplified products were amplified by PCR using the universal primers of 16s rRNA V6~V8 region, and the amplified products were subjected to denaturing gradient gel electrophoresis (DGGE). Cluster analysis, principal component analysis and variance analysis were used to analyze the structure of intestinal flora and its correlation with immune indexes. Results the composition of intestinal flora showed high polymorphism and significant individual difference. The levels of CD3 ~ T cells and CD4 ~ / CD8 ~ were correlated with intestinal microflora structure. The population with higher CD _ 4 ~ / CD _ 8 ~ was higher than those with lower CD _ 4 ~ / CD _ 8 ~ and normal population. Group richness (S) is low, However, there was no significant correlation between IgA IgMN IgG and intestinal flora structure. Conclusion the structure of intestinal flora is related to the immune level, and the decrease of the diversity of intestinal flora may affect the non-specific natural immune function centered on CD8T cells.
【作者单位】：四川大学华西公共卫生学院;河北医科大学公共卫生学院;
【基金】：达能营养中心膳食营养研究与宣教基金(No.DIC2014-01)
【分类号】：R392

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