口腔生物医学 ›› 2021, Vol. 12 ›› Issue (4): 242-256.

• 论著 • 上一篇    下一篇

颊纤毛菌对变异链球菌产酸及黏附作用的影响

高哲丰,孙宇,雷雅燕,李丹薇   

  1. 昆明医科大学附属口腔医院
  • 收稿日期:2021-09-24 修回日期:2021-12-10 出版日期:2021-12-25 发布日期:2022-01-04
  • 通讯作者: 李丹薇 E-mail:8942424@qq.com

Effect of Leptotrichia buccalis strain on acid production and adhesion of Streptococcus mutans

  • Received:2021-09-24 Revised:2021-12-10 Online:2021-12-25 Published:2022-01-04

摘要: 目的:研究分别使用颊纤毛菌(L. buccalis)及其上清液与变异链球菌(S. m)共培养,观察S. m的产酸、黏附变化及乳酸脱氢酶(ldh)、葡糖基转移酶(gtfb、gtfc、gtfd)基因表达变化。方法:首先从龈上菌斑中分离口腔L. buccalis,采用生化鉴定和测序鉴定;将L. buccalis单独培养组、S. m单独培养组、L. buccalis与S. m共培养组和S. m加入L. buccalis上清液培养组均培养9h,比较每小时各组细菌产酸能力的变化;再将L. buccalis单独培养组、S. m单独培养组、分别加入1、2、3 mL L. buccalis菌液的S. m共培养组和分别加入1、2、3 mL L. buccalis上清液的S. m菌液培养组,用实时定量RT-PCR法测定各组ldh、gtfb、gtfc和gtfd的基因表达;在S. m中分别加入50、100、150 μL L. buccalis上清液培养,在激光共聚焦显微镜下观察S. m黏附数量变化。结果:本实验成功分离L. buccalis,S. m中加入L. buccalis或L. buccalis上清液培养后pH下降速率增快,细菌的黏附数量增多,且随L. buccalis或L. buccalis上清液体积的增加ldh、gtfb、gtfc和gtfd的基因表达增高。结论:L. buccalis及其上清液对S. m的产酸及黏附能力具有促进作用。

关键词: 龋病, 牙菌斑生物膜, 颊纤毛菌上清液, 变异链球菌, 产酸

Abstract: Objective:To study the co?culture of Leptotrichia buccalisstrain (L.buccalis) and its supernatant with Streptococcus mutans (S.m) to observe the acid production, adhesion changes and ldh, gtfb, gtfc, gtfd gene expression changes. Methods: L.buccalis were isolated from the supragingival plaque and identified by biochemical identification and sequencing, L.buccalis culture group, S.mculture group, L.buccalis and S.m co?culture group, and S.m were added to the L.buccalis supernatant culture group and cultured for 9 hours, and the acid production capacity of each group was compared every hour. Then the L.buccalis culture group, the S.m culture group, the S.m co?culture with 1, 2 and 3 mL of L.buccalis culture group and add 1,2 and 3mL L.buccalis supernatant into S.m culture group to cultivate for 6 hours, and use real?timeRT?PCR method to determine and compare the gene expression of ldh, gtfb, gtfc and gtfd; Add 1,2,3 mL of L.buccalis supernatant to S.m andc o?culture for 9h, and the pH was measured every 1 hour. Compare the changes in acid production capacity; add 50,100 and 150 μL of L.buccalis supernatants to S.m and culture for 24h, and observe the changes in the adhesion number of S.m under a laser confocal microscope. Add 1,2,3mL of L.buccalis supernatant to S.m and culture for 6h,and use real?time RT?PCR to determine the gene expression of ldh, gtfb, gtfc and gtfd. Results: This experiment successfully isolated L.buccalis; After adding L.buccalis or L.buccalis supernatant to S.m, the pH value decreased faster, and the number of bacterial adhesions increased, and the volume of the L.buccalis or L.buccalis increased ldh, gtfb, gtfc and gtfd gene expression increased. Conclusions: L.buccalis and its supernatant can regulate the acid production and adhesion ability of S.m.

Key words: caries, dental plaque biofilm, L.buccalis supernatant, Streptococcus mutans, acid production