上海口腔医学 ›› 2019, Vol. 28 ›› Issue (4): 362-367.doi: 10.19439/j.sjos.2019.04.005

• 论著 • 上一篇    下一篇

赤藓糖醇对牙周致病菌抗菌性能的影响

张佳丽1, 姚军1, 诸葛继娜2, 张琰君1   

  1. 1.福建医科大学附属口腔医院 儿童口腔科, 福建 福州 350000;
    2.安徽省合肥市口腔医院 儿童口腔科, 安徽 合肥 230000
  • 收稿日期:2019-03-27 修回日期:2019-05-23 出版日期:2019-08-25 发布日期:2019-09-23
  • 通讯作者: 姚军,E-mail:dentyao@163.com
  • 作者简介:张佳丽(1984-),女,医师,硕士研究生,E-mail:zjl104009012@126.com
  • 基金资助:
    福建生卫计委医学创新课题(2017-cx-33)

Antibacterial activity of erythritol on periodontal pathogen

ZHANG Jia-li1, YAO Jun1, ZHUGE Ji-na2, ZHANG Yan-jun1   

  1. 1. Department of Pediatric Dentistry, Affiliated Stomatological Hospital of Fujian Medical University. Fuzhou 350000, Fujian Province;
    2. Department of Pediatric Dentistry, Hefei Stomatological Hospital. Hefei 230000, Anhui Province, China
  • Received:2019-03-27 Revised:2019-05-23 Online:2019-08-25 Published:2019-09-23

摘要: 目的 研究赤藓糖醇对牙周致病菌牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)、 伴放线聚集杆菌(Aggregatibacter actinomycetemcomitans, Aa)和粘性放线菌(Actinomyces viscous,Av)生长的影响,探讨不同浓度赤藓糖醇作用后的Pg对牙周膜细胞炎症相关细胞因子mRNA表达水平的影响。方法PgAaAv 3种致病菌接种于0、2、4、8、16、32、64、128 g/L的赤藓糖醇-BHI液中,37℃厌氧培养一定时间,检测其最低抑菌浓度。将Pg分别接种于MIC、1/2、1/4、1/8 MIC 4个赤藓糖醇质量浓度的培养基以及不含赤藓糖醇的培养基,离心并清洗后,加入细胞DMEM培养基重悬,与培养至第4代的人牙周膜细胞共培养24 h,弃上清,裂解细胞,提取总RNA,反转录,实时荧光定量PCR检测IL-1β、IL-6、TNF-α的mRNA相对表达量。采用SPSS19.0软件包对数据进行统计学分析。结果 赤藓糖醇对3种细菌的最低抑菌浓度分别为Pg:64 g/L,Aa:128 g/L,Av:128 g/L。不同浓度赤藓糖醇条件下培养的细菌,刺激牙周膜细胞产生IL-1β、IL-6 、TNF-α的能力不同。赤藓糖醇浓度为8 g/L时,炎症因子量与不加赤藓糖醇的对照组无显著差别;浓度升高达到16 g/L时,IL-1β、IL-6 、TNF-αmRNA的相对表达量有所降低,且浓度越高,炎症因子释放越少;但所有实验组炎症因子量始终高于未加细菌的空白对照组(P<0.05)。结论 赤藓糖醇对PgAaAv的生长能力有抑制作用,且在一定范围内,赤藓糖醇质量浓度越高,抑制效果越明显。赤藓糖醇还可通过某种方式对致病菌毒力因子起抑制作用,进而降低Pg的牙周致炎性,减少牙周膜细胞IL-1β、IL-6、TNF-α的mRNA相对表达量。

关键词: 牙龈卟啉单胞菌, 伴放线聚集杆菌, 粘性放线菌, 赤藓糖醇, 牙周炎症因子

Abstract: PURPOSE: To explore the effect of erythritol on the growth of Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans(Aa), Actinomices viscosus (Av), and to explore how Porphyromonas gingivalis affected by erythritol influence mRNA expression level of inflammatory in periodontal cells. METHODS: Pg, Aa, Av were anaerobically cultured (80%N2, 10%CO2, 10%H2) at 37℃ in 2, 4, 8, 16, 32, 64, 128 g/L erythritol- BHI mixture groups (experimental groups) and BHI groups (control group). The lowest erythritol concentration without turbidity or precipitation was the minimum inhibitory concentration. Pg was cultured in MIC, 1/2, 1/4, 1/8 MIC erythritol- BHI mixture groups (experimental groups) and BHI groups (control group). Each kind of bacteria in each concentration group was centrifugalled and cleaned before added into DMEM. The mixed suspension was co-cultured with the periodontal ligament cells in four generations for 24 hours, the supernatan was removed , then the total RNA in cracking cells was extracted and reversing transcription. At last, the relative expression of IL-1, IL-6, TNF-a was detected real-time fluorescent quantitative PCR. The data were analyzed with SPSS19.0 software package. RESULTS: The minimum inhibitory by concentrations of erythritol on three bacteria were as followed: Pg: 64 g/L,Aa: 128 g/L,Av: 128 g/L. The ability of stimulating periodontal ligament cells to produce IL-1β, IL-6 and TNF-α was different when Pg was cultured under different concentrations of erythritol. There was no significant difference between 0 g/L of control group and 8 g/L of experimental group. As the concentration reached 16 g/L, the relative expression of IL-1β, IL-6, TNF-α was reduced, and the higher concentration was, the less inflammatory factors was. However, the inflammatory factors in all the experimental groups were always significantly higher than that in the blank control group (P<0.05). CONCLUSIONS: Erythritol has an inhibitory effect on the growth of Pg, Aa and Av. In a certain range, higher concentration of erythritol delivers better inhibition effect. Erythritol can also reduce the periodontal pathogenicity of pathogenic bacteria in a way inhibiting the virulence of these bacteria, reducing the production of IL-1β, IL-6, and TNF-a in periodontal cells.

Key words: Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Actinomyces viscous, Erythritol, Periodontal pathogenicity

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