基于高通量测序分析鲜槟榔水提液对于小鼠肠道菌群及免疫指标的影响

High-throughput Sequencing Analysis of Fresh Areca Nut Extract on Intestinal Flora and Immune Indexes of Mice

通过给予小鼠不同剂量鲜槟榔水提液(fresh areca nut water extract,FANE),记录每组小鼠体重增长、分析其血清细胞因子白介素-1 (interleukin 1,IL-1)、α肿瘤坏死因子(tumor necrosis factorα,TNF-α),免疫球蛋白A(immunoglobin A,IgA)、IgG与IgM水平,并通过高通量测序分析结肠微生物菌群的多样性与丰度变化,以探究槟榔灌胃小鼠免疫指标变化与小鼠肠道微生物菌群结构的关系。结果表明,低剂量FANE可显著提高小鼠IL-1的浓度,并降低其IgA和IgG的水平。高剂量FANE造成了小鼠TNF-α的显著降低以及IL-1浓度的显著增加,减少了血清IgA、IgG并提高了其IgM水平。这说明高剂量FANE可能造成了小鼠免疫功能的紊乱。对于小鼠肠道菌群的Alpha多样性分析显示,低剂量组(A1)与对照组(F)具有较相似的肠道菌群结构与菌群丰度和多样性,而高剂量组(A2)相较其它两组肠道菌群多样性明显较低。具体分析,A2组其门水平上厚壁菌门(Firmicutes)与拟杆菌门(Bacteroidetes)比例明显增加,这可能与食物中能量利用率的改变相关。属水平上阿克曼氏菌(Akkermansia)、Enterorhabdus菌与葡萄球菌(Staphylococcus)的显著增加可能反映了肠道炎症与感染的发生以及Zn的缺乏。此外,该组小鼠的长期腹泻也可能促进了上述结果的出现。这些分析表明,肠道微生物结构的显著变化可能在小鼠免疫指标与功能改变过程中起到重要的作用。

The purpose of this study was to explore the relationship between the changes of immune indexes and the structure of intestinal microflora in fresh areca nut water extract(FANE)gavage mice. Serum cytokine interleukin 1(IL-1),alpha tumor necrosis factor(TNF-α),immunoglobulin A(IgA),IgG and IgM level of each group of mice were analyzed and weight gain were calculated. Besides,the abundance of colonic microflora was analyzed by high throughput sequencing. The results showed that low-dose FANE significantly increased the concentration of IL-1 and decreased the levels of IgA and IgG. The administration of high-dose FANE resulted in a significant decrease in TNF-α,serum IgA and IgG,and a significant increase in IL-1 concentration and IgM level. This indicated that high-dose FANE could induce immune dysfunction in mice. Alpha diversity analysis of mice intestinal flora showed that low-dose FANE group(A1)and the control group(F)had similar intestinal flora abundance and diversity. While the diversity of intestinal flora in the high-dose group(A2)was much lower than that in the other two groups. The ratio of Firmicutes/Bacteroidetes in the phylum level of A2 group increased significantly,which may be related to the change of energy utilization in food. The significant increase of Akkermansia,Enterorhabdus and Staphylococcus on the genus level may reflect the occurrence of intestinal inflammation and infection as well as Zn deficiency. In particular,chronic diarrhea in this group of mice might have contributed to these results. These analyses suggested that significant changes in gut microbial structure may play an important role in the changes in immune indices and function in mice.