应用16S rDNA扩增子序列分析去卵巢骨丢失小鼠口腔菌群的变化

Oral microbiota changes of bone loss mice induced by ovariectomy via 16S rDNA amplicon sequencing

  • 摘要:
      背景  口腔微生物失调加重了多种口腔疾病和系统性疾病的进程。进入绝经后期的妇女罹患口腔黏膜病和加速牙周骨质丢失的风险更高,但口腔菌群变化与女性进入绝经后期之间的联系尚不清楚。
      目的  应用16S rDNA扩增子测序,初步观察卵巢切除小鼠的口腔菌群结构变化。
      方法  16只雌性C57BL/6J小鼠随机分为卵巢切除(ovariectomy,OVX)组和假手术(sham-operated,Sham)组。卵巢切除术4周后对股骨进行显微CT扫描和骨密度与骨参数的分析,取小鼠口腔拭子进行16S rDNA高通量测序。
      结果  与Sham组比较,OVX组骨密度、骨体积分数、骨小梁厚度和骨小梁数量显著下降,骨表面积/骨体积和骨小梁分离度显著升高(P<0.05)。两组的口腔菌群共同含有687种ASVs,Sham组特有1086种,OVX特有930种。OVX组的Chao1指数与Sham组差异无统计学意义,但Simpson指数显著低于Sham组(P<0.05)。门水平上OVX组的拟杆菌门和梭杆菌门丰度较Sham组下降(P<0.05)。属水平上OVX组的梭杆菌属、乳酸乳球菌属、萨特氏菌属等菌属的丰度相比SHAM组降低(P<0.05);放线菌属等菌属的丰度相比Sham组升高(P<0.05)。LEfSe分析结果显示,Sham组厚壁菌门和芽孢杆菌纲(Bacilli)的相对丰度升高(P<0.05)。
      结论  卵巢切除小鼠出现骨丢失现象,并且口腔菌群的多样性和菌群结构发生了改变,初步为研究绝经后妇女口腔菌群改变与口腔疾病加重的关系提供依据。

     

    Abstract:
      Background  The dysbiosis of oral microorganisms aggravates the process of several oral diseases and systemic diseases. Postmenopausal women are at increased risk of oral mucosal disease and accelerated periodontal bone loss, but the association between changes in oral microbiota and postmenopause in women is unclear.
      Objective  To explore the changes of oral microbiota in ovariectomized mice via 16S rDNA amplicon sequencing.
      Methods  Sixteen female C57BL/6J mice were randomly divided into Ovariectomy group (OVX) and sham-operated group (Sham). The femurs were taken for MicroCT scanning and bone mineral density and bone parameters analysis at 4 weeks after the operation, and the oral swabs samples were collected for 16S rDNA high-throughput sequencing.
      Results  The bone mineral density, bone volume/total volume, trabecular thickness and trabecular number were decreased, while the bone surface area/bone volume and bone trabecular spacing were increased in OVX group (P<0.05). Two groups of oral samples contained common 687 ASVs, OVX group contained specific 1086 ASVs and OVX group contained specific 930 ASVs. There was no significant difference in Chao1 index between OVX group and Sham group, Simpson index in OVX group was significantly lower than that in Sham group (P<0.05). At the phylum level of oral samples, the relative abundance of Fusobacteriota and Bacteroidota in OVX group was decreased (P<0.05). At the genus level, the relative abundance of Fusobacterium, Lactococcus, Sutterella and other genera in OVX group were decreased, Actinomyces and other genera were increased (P<0.05). LEfSe analysis showed that the relative abundance of Firmicutes and Bacilli increased in the Sham group (P<0.05).
      Conclusion  Bone loss occurs in ovariectomized mice, and the diversity and structure of oral microbiota are changed, which preliminarily provides reference for the study of the relationship between the change of oral microbiota and the aggravation of oral diseases in postmenopausal women.

     

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