Journal of Prevention and Treatment for Stomatological Diseases ›› 2021, Vol. 29 ›› Issue (5): 314-321.DOI: 10.12016/j.issn.2096-1456.2021.05.004

• Basic Study • Previous Articles     Next Articles

Inhibitory effects of epigallocatechin-3-gallate on the pathogenic properties of P. gingivalis in vitro

QI Xia1(),KONG Lingxue2,LI Shujuan1,MA Siting1,QI Yali1,ZHAO Lei3()   

  1. 1. Department of Periodontics, School and Hospital of Stomatology, Hebei Medical University, Hebei Key Laboratory of Stomatology,Hebei Clinical Research Center for Oral Diseases, Shijiazhuang 050017, China
    2. Department of Periodontics and Oral Mucosa, Jinan Stomatologic Hospital, Jinan 250001, China
    3. State Key Laboratory of Oral Diseases & National Clinical Research Center for Oral Diseases, Department of Periodontics, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, China
  • Received:2020-11-13 Revised:2020-12-13 Online:2021-05-20 Published:2021-03-08
  • Contact: Lei ZHAO
  • Supported by:
    Medical Science Research Project in Hebei Province(20190096);Natural Science Foundation of Hebei Province(H2019206541);Senile Disease Prevention project of Hebei Provincial Department of Finance(2017068877-2)

表没食子儿茶素没食子酸酯对牙龈卟啉单胞菌细菌毒力抑制作用的体外研究

齐霞1(),孔令雪2,李淑娟1,马思婷1,齐雅丽1,赵蕾3()   

  1. 1.河北医科大学口腔医学院·口腔医院牙周一科,河北省口腔医学重点实验室,河北省口腔疾病临床医学研究中心,河北 石家庄(050017)
    2.济南口腔医院牙周黏膜科,山东 济南 (250001)
    3.口腔疾病研究国家重点实验室 国家口腔疾病临床医学研究中心 四川大学华西口腔医院牙周科,四川 成都(610041)
  • 通讯作者: 赵蕾
  • 作者简介:齐霞,主治医师,硕士,Email: summyly@163.com
  • 基金资助:
    河北省医学科学研究课题计划(20190096);河北省自然科学基金项目(H2019206541);河北省财政厅老年病防治项目(2017068877-2)

Abstract:

Objective To explore the antibacterial activity of epigallocatechin-3-gallate (EGCG) on P. gingivalis and the inhibitory effects on matrix metalloproteinases (MMPs) production induced by P. gingivalis. Methods The antimicrobial effect of EGCG against planktonic cultures and biofilms of P. gingivalis was evaluated using microplate dilution assays. The microstructural changes in biofilms were studied using scanning electron microscopy (SEM). The inhibitory effect of EGCG on arginine gingipain (Rgp) and lysine gingipain (Kgp) activity of P. gingivalis was evaluated using synthetic chromogenic peptides and fluorogenic substrates. Enzyme-linked immunosorbent assay (ELISA) and qRT-PCR analysis were used to assess MMP-1 and MMP-2 mRNA expression and secretion by human gingival fibroblasts (HGFs) stimulated with P. gingivalis in the presence or absence of EGCG, respectively. Results The MIC and MBC of EGCG against P. gingivalis were 62.5 μg/mL and 500 μg/mL, respectively. EGCG can not only inhibit the biofilm formation of P. gingivalis but also has a scavenging effect on mature biofilms and can affect their viability. Additionally, 10 μg/mL and 50 μg/mL of EGCG inhibited the proteinase activities of Rgp and Kgp, respectively (P < 0.05). Finally, the mRNA expression and secretion of MMP-1 and MMP-2 by HGFs stimulated by P. gingivalis were significantly inhibited by 50 μg/mL of EGCG (P < 0.05). Conclusion EGCG exhibits antimicrobial effects against P. gingivalis and reduces the expression of MMPs by HGFs.

Key words: epigallocatechin-3-gallate, Porphyromonas gingivalis, biofilm formation, biofilm reduction, arginine gingipain, lysine gingipain, antibacterial, anti-inflammatory, gingival fibroblasts, matrix metalloproteinase

摘要:

目的 研究表没食子儿茶素没食子酸酯(epigallocatechin-3-gallate,EGCG)对牙龈卟啉单胞菌(Porphyromonas gingivalis,P. gingivalis)体外抑菌活性及EGCG对P. gingivalis诱导人牙龈成纤维细胞(human gingival fibroblasts,HGFs)表达基质金属蛋白酶(matrix metalloproteinases,MMPs)的抑制作用。方法 采用微量稀释法分析EGCG对P. gingivalis浮游菌和生物膜的抑制作用。扫描电镜检测EGCG作用下P. gingivalis生物膜微观结构的变化。特异性人工合成显色多肽底物评价EGCG对牙龈素精氨酸特异性半胱氨酸蛋白酶 (arginine gingipain, Rgp) 和赖氨酸特异性半胱氨酸蛋白酶 (lysine gingipain, Kgp)蛋白水解活性的影响。P. gingivalis刺激HGFs,与EGCG共培养,ELISA检测MMP-1和MMP-2的蛋白分泌水平,qRT-PCR检测MMP-1及MMP-2 mRNA的表达。结果 EGCG对P. gingivalis浮游菌的最低抑菌浓度为62.5 μg/mL,最低杀菌浓度为500 μg/mL。EGCG可抑制P. gingivalis生物膜形成,并对成熟生物膜具有清除作用,且抑制成熟生物膜的代谢活性。10 μg/mL及50 μg/mL EGCG可有效抑制牙龈素的水解活性(P < 0.05)。此外,50 μg/mL EGCG对P. gingivalis刺激HGFs表达及分泌MMP-1、MMP-2均有抑制作用(P < 0.05)。结论 EGCG对P. gingivalis有抑菌作用,且可抑制HGFs表达MMPs。

关键词: 表没食子儿茶素没食子酸酯, 牙龈卟啉单胞菌, 生物膜形成, 生物膜清除, 精氨酸特异性半胱氨酸蛋白酶, 赖氨酸特异性半胱氨酸蛋白酶, 抗菌, 抗炎, 牙龈成纤维细胞, 基质金属蛋白酶

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