摘要
目的探讨新疆桑白皮防龋潜在机制,并分析其对主要致龋菌的作用。方法TCMSP数据库筛选新疆桑白皮活性成分及靶点。Genecards、DisGENET和TTD数据库获取龋病靶点。获取新疆桑白皮防龋的共同靶点,并筛选核心基因。使用DAVID数据平台进行富集分析。采用AutoDock软件进行分子对接验证。通过体外抑菌实验,首先测定新疆桑白皮提取物对变异链球菌、血链球菌、黏性放线菌的50%最低抑菌浓度(50%minimum inhibitory concentration,MIC50)、最低杀菌浓度(minimal bactericidal concentration,MBC),并绘制生长曲线;分别测定新疆桑白皮提取物对变异链球菌、血链球菌、黏性放线菌浮游状态产酸、产糖和黏附能力的影响;通过结晶紫染色法测定新疆桑白皮提取物对主要致龋细菌单菌种50%最低生物膜抑制浓度(50%minimum biofilm inhibition concentration,MBIC50)和50%最低已形成生物膜清除浓度(50%minimum biofilm re-duction concentration,MBRC50),并利用扫描电子显微镜观察生物膜形态。结果筛选出新疆桑白皮防龋关键活性成分包括槲皮素、山奈酚、β-谷甾醇等。肿瘤坏死因子(tumor necrosis factor,TNF)、白细胞介素-6(interleu-kin-6,IL-6)、白细胞介素-1β(interleukin-1beta,IL-1β)等可能是新疆桑白皮防龋的关键靶点。富集分析结果显示新疆桑白皮可能对AGE-RAGE信号通路、IL-17信号通路、TNF信号通路等产生影响。抑菌实验结果显示新疆桑白皮提取物对变异链球菌、血链球菌、黏性放线菌的MIC50分别为0.5、0.5、0.25 mg/mL,MBC分别为4.0、2.0、1.0 mg/mL。新疆桑白皮提取物对3种主要致龋细菌浮游状态产酸、产多糖以及黏附能力的抑制作用较对照组强,差异均具有统计学意义(P<0.05)。新疆桑白皮提取物对变异链球菌、血链球菌、黏性放线菌的MBIC50分别为1.0、1.0、0.5 mg/mL,MBRC50分别为4.0、4.0、2.0 mg/mL。扫描电镜结果显示随着新疆桑白皮药物浓度的增加,生物膜形成量与对照组相比逐渐减少。结论新疆桑白皮可通过槲皮素、山奈酚、β-谷甾醇等活性成分,TNF、IL-6、IL-1β等关键靶点及多种作用途径防龋,不仅能抑制主要致龋细菌浮游状态生长、产酸、产糖、黏附能力且对致龋菌生物膜形成有一定的抑制作用。
Objective To investigate the potential caries prevention mechanism of the Xinjiang Mori cortex and to analyze its effect on the main cariogenic bacteria.Methods The active components of the Xinjiang Mori cortex and the main targets were predicted and screened using the TCMSP database.The GeneCards,DisGENET and TTD databas⁃es were used to obtain caries⁃related targets.The common targets were derived,and core genes were screened.The en⁃richment analysis was performed using the DAVID data platform.Molecular docking was performed using AutoDock software.In in vitro antibacterial experiments,first,the 50%minimum inhibitory concentration(MIC50)and the mini⁃mum bactericidal concentration(MBC)of the Xinjiang Mori Cortex extract against Streptococcus mutans,Streptococcus sanguis and Actinomyces viscosus were determined and the growth curves were measured.The effects of the Xinjiang Mo⁃ri Cortex extract on acid production,polysaccharide production and adhesion ability of Streptococcus mutans,Streptococ⁃cus sanguis and Actinomyces viscosus in the planktonic state were determined.The 50%minimum biofilm inhibition con⁃centration(MBIC50)and 50%minimum biofilm reduction concentration(MBRC50)were determined by crystal violet stain⁃ing,and biofilm morphology was visualized using scanning electron microscopy(SEM).Results The main active com⁃ponents of the Xinjiang Mori cortex included quercetin,kaempferol,andβ⁃sitosterol.Tumor necrosis factor(TNF),inter⁃leukin⁃6(IL⁃6),and interleukin⁃1beta(IL⁃1β)could be the most important targets of the Xinjiang Mori cortex for the prevention of dental caries.The enrichment analysis results showed that Mori cortex extract may have effects on the AGE⁃RAGE signaling pathway,IL⁃17 signaling pathway,and TNF signaling pathway.The antibacterial experiment re⁃sults showed that the MIC50 values of Xinjiang Mori Cortex extract against Streptococcus mutans,Streptococcus sanguis and Actinomyces viscosus were 0.5,0.5 and 0.25 mg/mL,respectively,and the MBCs were 4.0,2.0 and 1.0 mg/mL,re⁃spectively.The inhibitory effect of Xinjiang Mori Cortex extract on the acid production,polysaccharide production and adhesion ability of three major cariogenic bacteria in the planktonic state was stronger than that of the control group,and the differences were statistically significant(P<0.05).The MBIC50 was 1.0,1.0,and 0.5 mg/mL,and the MBRC50 was 4.0,4.0,and 2.0 mg/mL.SEM observation showed that the amount of biofilm formation decreased with the drug concen⁃tration compared with the control group.Conclusion Xinjiang Mori cortex extract can prevent caries through querce⁃tin,kaempferol,andβ⁃sitosterol active ingredients,TNF、IL⁃6、IL⁃1βkey targets and multiple pathways and inhibit the growth,acid production,polysaccharide production,and adhesion ability of three major cariogenic bacteria in the plank⁃tonic state and has some inhibitory effect on corticogenic biofilm formation.
作者
阿尔曼·阿卜力孜
吴龙
玛依奴尔·阿塔吾拉
赵今
AERMAN Abulizi;WU Long;MAYINUER Atawula;ZHAO Jin(Depart-ment of Cariology and Endodontics,The First Affiliated Hospital of Xinjiang Medical University(Affiliated Stomatology Hospital),Urumqi 830054,China;Stomatology Disease Institute of Xinjiang Uygur Autonomous Region,Urumqi 830054,China)
出处
《口腔疾病防治》
2024年第2期89-100,共12页
Journal of Prevention and Treatment for Stomatological Diseases
基金
新疆维吾尔自治区自然科学基金(2022D01C751)
新疆医科大学第一附属医院“青年科研起航”专项基金(2022YFY⁃QKQN⁃40)。
关键词
网络药理学
分子对接
天然药物
桑白皮
提取物
龋病
防龋
致龋细菌
抑菌
毒力因子
network pharmacology
molecular docking
natural medicine
Mori Cortex
extract
caries
caries prevention
cariogenic bacteria
bacteriostasis
virulence factor
