牛至精油对变形链球菌的体外抑菌活性及活性成分分析,BMC Complementary Medicine and Therapies

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牛至精油对变形链球菌的体外抑菌活性及活性成分分析
BMC Complementary Medicine and Therapies Pub Date : 2023-02-21 , DOI: 10.1186/s12906-023-03890-4
Yue Yuan , Jinlong Sun , Yang Song , Rifat Nowshin Raka , Jie Xiang , Hua Wu , Junsong Xiao , Jianming Jin , XiuLi Hui

背景

变形链球菌( S. mutans ) 被认为是在非致病性共生口腔微生物群落向促进龋齿发展的菌斑生物膜转变过程中最相关的细菌。牛至（Origanum vulgare L. ）是一种普遍存在的天然调味品，其精油已被证明具有良好的抗菌作用。然而，牛至精油 (OEO) 对变形链球菌的具体抗菌机制仍未完全了解。

方法

在这项工作中，两种不同的 OEO 的组成由 GC-MS 确定。确定纸片扩散法、最低抑菌浓度 (MIC) 和最低杀菌浓度 (MBC) 以评估它们对变形链球菌的抗菌作用。变形链球菌对gtfB / C / D、spaP、gbpB、vicR、relA和brpA mRNA 表达的产酸、疏水性、生物膜形成和实时 PCR 的抑制进行了评估以初步调查其作用机制。进行分子对接以模拟与毒力蛋白和活性成分的相互作用。还进行了使用永生化人角质形成细胞的 MTT 测试以研究细胞毒性。

结果

与阳性药物青霉素/链霉素 100X（DIZ：34.13±0.85 mm，MIC：0.78125 μL/mL，MBC：6.25 μL/mL）相比，牛至的精油。（DIZ：80 mm，MIC：0.625μL/mL，MBC：2.5μL/mL）和Origanum heracleoticum L。（DIZ：39.67 ± 0.81 mm，MIC：0.625μL/mL，MBC：1.25μL/mL）在 1/2-1MIC 浓度下也可以表现出类似的抑制产酸和降低变形链球菌的疏水性和生物膜形成的作用. 以及gtfB/C/D、spaP、gbpB、vicR和relA的基因表达被发现被下调。由于不同来源的精油成分差异很大，通过有效的网络药理学分析，我们发现 OEOs 含有许多有效化合物，如香芹酚及其生物合成前体 γ-萜品烯和对伞花烃，它们可能直接针对多种毒力蛋白变形链球菌。此外，在永生化人角质形成细胞中，0.1 μL/mL 的 OEO 没有引起毒性作用。

结论

本研究的综合分析表明，OEO 可能是一种潜在的预防龋齿的抗菌剂。

"点击查看英文标题和摘要"

Antibacterial activity of oregano essential oils against Streptococcus mutans in vitro and analysis of active components

Background

Streptococcus mutans (S. mutans) is considered the most relevant bacteria during the transition of the non-pathogenic commensal oral microbial community to plaque biofilms that promote the development of dental caries. Oregano (Origanum vulgare L.), is a universally natural flavoring and its essential oil has been demonstrated to have good antibacterial effects. However, the specific antibacterial mechanism of oregano essential oil (OEO) against S. mutans is still not completely understood.

Methods

In this work, the composition of two different OEOs was determined by GC‒MS. Disk-diffusion method, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined to assess their antimicrobial effect on S. mutans. The inhibition of acid production, hydrophobicity, biofilm formation and real-time PCR for gtfB/C/D, spaP, gbpB, vicR, relA and brpA mRNA expression by S. mutans were assessed to preliminarily investigate the mechanisms of action. Molecular docking was performed to simulate the interactions with the virulence proteins and active constituents. MTT test using immortalized human keratinocytes cells was also performed to investigate cytotoxicity.

Results

Compared with the positive drug Penicillin /streptomycin 100X (DIZ: 34.13 ± 0.85 mm, MIC: 0.78125 μL/mL, MBC: 6.25 μL/mL), the essential oils of Origanum vulgare L. (DIZ: 80 mm, MIC: 0.625μL/mL, MBC:2.5μL/mL) and Origanum heracleoticum L. (DIZ: 39.67 ± 0.81 mm, MIC: 0.625μL/mL, MBC: 1.25μL/mL) could also exhibit similar effects to inhibit the acid production and reduce the hydrophobicity and biofilm formation of S. mutans at 1/2-1MIC concentration. And gene expression of gtfB/C/D, spaP, gbpB, vicR and relA were found to be downregulated. Due to the composition of essential oils from different sources being highly variable, through effective network pharmacology analysis, we found that OEOs contained many effective compounds, like carvacrol and its biosynthetic precursors γ-terpinene and p-cymene, which may directly target several virulence proteins of S. mutans. Besides, no toxic effect was instigated by OEOs at 0.1 μL/mL in the immortalized human keratinocytes cells.