兔膝骨性关节炎模型肠道菌群的16S rRNA检测分析△

16S rRNA detection and analysis of gut microbiota in osteoarthritis model in rabbits

摘要:

［目的］ 探讨肠道菌群在兔膝骨关节炎 （knee osteoarthritis, KOA） 与正常对照之间的差异。［方法］ 20 只大耳白兔， 随机分成 2 组，12 只关节内注射 2％木瓜蛋白酶溶液制备 KOA 模型，另外 8 只关节内注射等量生理盐水为正常对照。采集动物粪便样本，利用 16S rRNA 基因靶向测序技术进行检测，分析各自粪便中肠道菌群的多样性及分布情况。［结果］ KOA 组粪便含有 20 188 种物种（ASV/OTU），而对照组含有 14 029 种（ASV/OTU）。KOA 组在粪便菌群的门、属水平上显著丰富于照组 （P<0.05），但两组在菌群纲、目、科、种水平上的差异无统计学意义 （P>0.05）。KOA 组在门水平以厚壁菌、拟杆菌的数量最多，在属水平上以瘤胃球菌属及颤螺菌属数量最多；其中明显的标志物种为变形杆菌。菌群主要与代谢通路密切相关，其中在碳水化合物代谢、氨基酸代谢及辅助因子和维生素代谢上极为突出。［结论］ KOA 组与对照组的肠道菌群之间存在一定差异， 标志物种为变形杆菌。

关键词:16S rRNA;肠道菌群;变形杆菌;膝骨关节炎

Abstract:

[Objective] To investigate the difference of gut microbiota between knee osteoarthritis (KOA) and normal control in rabbits. [Methods] A total of 20 Japanese white rabbits were randomly divided into 2 groups, of them, 12 animals were injected 2% papain solution intraarticularly to prepare KOA model, while the other 8 rabbits were injected normal saline intraarticularly as normal control. Fecal sam- ples were collected and detected by 16S rRNA gene targeted sequencing technology to analyze the diversity and distribution of gut microbio- ta in their feces. [Results] The KOA group had a total of 20 188 species (ASV/OTU) , while the control group was of 14 029 species (ASV/ OTU) . The fecal flora in KOA group was significantly more abundant than that in the control group in levels of phylum and genus (P< 0.05) , but there were no statistical significant differences in the levels of class, order, family and specie between the two groups (P>0.05) . The KOA group had the greatest number of firmicutes and bacteroides at phylum level, and ruminococcus and spironella flutter at genus lev- el, with an obvious marker species of proteus. The gut microbiota was closely related to metabolic pathways, especially carbohydrate metab- olism, amino acid metabolism, cofactor and vitamin metabolism. [Conclusion] There are some differences in gut microbiota between KOA group and control group, and proteus is the marker species.

Key words:16S rRNA; gut microbiota; proteus; knee osteoarthritis