基于高通量测序的壳寡糖-Zn^2＋配合物对衰老模型小鼠肠道菌群结构的影响

Effect of chitooligosaccharides-Zn^(2+) on intestinal microbiota in induced aging mice based on Miseq high-throughput sequencing analysis

考察壳寡糖-Zn^(2+)配合物对D-半乳糖诱导衰老小鼠肠道菌群结构的影响。以D-半乳糖注射法制备氧化衰老模型小鼠,设立空白、模型对照、阳性对照(V_C)、壳寡糖、Zn SO_4、壳寡糖+Zn SO_4和壳寡糖-Zn^(2+)配合物组,持续给药30 d后,取小鼠盲肠部分进行肠道菌群分析。结果发现,各样本有效序列范围为31727~36486条、OTU个数范围为255~376,其中空白组OTU数较低、壳寡糖及其配合物组OTU数量较高。在门水平,D-半乳糖显著影响(p<0.05)空白组厚壁菌门、拟杆菌门丰度及相互关系,壳寡糖和壳寡糖-Zn^(2+)配合物对模型小鼠厚壁菌门、拟杆菌门具有一定恢复作用。在科水平,相比于正常组,模型组Bacteroidales_S24-7_group丰度大幅增加,而壳寡糖-Zn^(2+)配合物给药后Bacteroidales_S24-7_group丰度值显著降低(p<0.05),同时其对消化球菌科、内瘤胃菌科及毛螺菌科丰度也具有一定的提升作用。与模型组相比,壳寡糖-Zn^(2+)配合物提高了样本Ace、Chao和Shannon指数,同时降低了Simpson指数。聚类和主成分分析(Principal Component Analysis,PCA)发现,壳寡糖-Zn^(2+)配合物与V_C给药组相比,肠道微生物相似性无显著性差异(p>0.05),且能使肠道菌群比例和分布更加接近于空白组样本。D-半乳糖诱导显著影响小鼠内肠道菌群的丰度及结构组成,而壳寡糖-Zn^(2+)配合物对模型小鼠肠道菌群结构具有一定逆向恢复作用。

The purpose of this study was to investigate the effects of ehitooligosaccharides-Zn^2＋ on intestinal microbiota in induced aging mice based on Miseq high-throughput sequencing analysis.In this study, the model of mice with induced aging were established with D- galactose injection. Then, the mice were randomly divided into seven groups, including the blank control, the model group, the groups treated with chitooligosaccharides, ZnSO4, chitooligosaccharides plus ZnSO4 and chitooligosaccharides-Zn^2＋ respectively.The results indicated that the numbers of valid sequence and OTU in different groups were ranged from 31727 to 36486 and from 255 to 376, respectively,in which the blank control showed the less OTU number, while the chitooligosaccharides and chitooligosaccharides-Zn^2＋ groups showed higher OTU numbers.At the phylum level, the abundances and rates of Firmicutes amd Bacteroidetes were significantly affected by the D- galactoseinjection, while the treatments of chitooligosaccharides and chitooligosaccharides-Zn2＋ showed the positive roles on the above two phylum.At the family level ,the abundance of BacteroidalesS24-7group was significantly（p 〈 0.05 ）increased in the model groups, but the chitooligosaccharides - Zn^2＋ decreased its abundance, which also increased the abundances of Peptococcaccae, Bacteriodesruminocola, and Lachnospiraceae compared with the model group. Additionally, the chitooligosaccharides- Zn^2＋ significantly（p 〈0.05）increased the index of Ace, Chao and Shannon and decreased the index of Simpson compared with the model group.The results of cluster and PCA analysis indicated there were no significant difference （p 〉 0.05 ） between the Vc and chitooligosaccharides-Zn^2＋ group, the proportion and distribution of which were close to the blank control group. In conclusion,the bacterial diversity and species abundance of intestinal microbiota in mice was significantly affected by the injection of D- galactose, while the treatment of chltoohgosaccharldes- Zn^2＋ showed the therapeutic effects on the intestinal microbiota in induced aging mice.