研究报告

YHX系列抗菌肽对典型口腔致病菌的抗菌作用及安全性评价

  • 张诗淇 ,
  • 刘伊索 ,
  • 刘银雪 ,
  • 曹佳媛 ,
  • 蒋璐 ,
  • 易华西
展开
  • (中国海洋大学 食品科学与工程学院,山东 青岛,266000)
第一作者:硕士研究生(易华西教授为通信作者,E-mail:yihx@ouc.edu.cn)

收稿日期: 2025-09-26

  修回日期: 2025-11-06

  网络出版日期: 2026-07-17

基金资助

国家重点研发计划国际合作项目(2024YFE111400)

Antibacterial effects and safety evaluation of YHX series antimicrobial peptides against typical oral pathogens

  • ZHANG Shiqi ,
  • LIU Yisuo ,
  • LIU Yinxue ,
  • CAO Jiayuan ,
  • JIANG Lu ,
  • YI Huaxi
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  • (College of Food Science and Engineering, Ocean University of China, Qingdao 266000, China)

Received date: 2025-09-26

  Revised date: 2025-11-06

  Online published: 2026-07-17

摘要

口腔健康易受多种内源性与外源性因素影响,其中口腔致病微生物是主要因素之一。抗菌肽因具有高效抗菌活性与独特作用机制,在口腔致病菌感染防治中显示出广阔应用前景。该研究系统评价了YHX系列人工合成抗菌肽对变异链球菌、具核梭杆菌和牙龈卟啉单胞菌这3种典型口腔致病菌的抑制作用。通过测定抑菌圈直径、最小抑菌浓度(minimum inhibitory concentration, MIC)、最小杀菌浓度(minimum bactericidal concentration, MBC)及最小生物膜抑制浓度(minimum biofilm inhibitory concentration, MBIC),评估其抗菌与抗生物膜活性;并利用扫描电镜(scanning electron microscopy, SEM)观察细菌生物膜形态变化,同时通过RAW264.7细胞毒性及溶血实验评价其生物安全性。结果表明,6种抗菌肽对变异链球菌和具核梭杆菌均表现出良好抗菌活性,MIC值为4~32 μg/mL,MBIC值均低于64 μg/mL。在4×MIC浓度下,该系列抗菌肽对变异链球菌生物膜的抑制率超过60%,对具核梭杆菌与牙龈卟啉单胞菌共培养生物膜的抑制率均高于85%。扫描电镜显示,抗菌肽可引起细菌细胞结构变形甚至破裂,从而发挥抑菌作用。细胞毒性实验表明,YHX-5和YHX-7具有较低的细胞毒性;溶血实验结果进一步证实YHX-5具有低溶血率和良好的细胞选择性。综上所述,YHX-5兼具高效抗菌活性、显著抗生物膜能力、低溶血毒性与低细胞毒性,展现出作为新型口腔抗菌剂的开发潜力。

本文引用格式

张诗淇 , 刘伊索 , 刘银雪 , 曹佳媛 , 蒋璐 , 易华西 . YHX系列抗菌肽对典型口腔致病菌的抗菌作用及安全性评价[J]. 食品与发酵工业, 2026 , 52(12) : 98 -105 . DOI: 10.13995/j.cnki.11-1802/ts.044621

Abstract

Oral health is influenced by various endogenous and exogenous factors, with pathogenic microorganisms representing a major contributor.Antimicrobial peptides (AMPs) demonstrate considerable potential for preventing and treating oral infections due to their potent antibacterial properties and unique mechanisms of action.This study systematically evaluated a series of synthetic YHX AMPs for their inhibitory effects against three common oral pathogens, including Streptococcus mutans, Fusobacterium nucleatum, and Porphyromonas gingivalis.The assessment included inhibition zone diameter, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and minimum biofilm inhibitory concentration (MBIC) to determine antibacterial and anti-biofilm activity.Scanning electron microscopy (SEM) imaging revealed structural alterations in bacterial biofilms.Biosafety evaluation employed cytotoxicity assays using RAW264.7 cells alongside haemolysis tests.Results indicated that all six YHX AMPs exhibited strong antibacterial activity against S.mutans and F.nucleatum.MIC values ranged from 4 to 32 μg/mL, while all MBIC values measured below 64 μg/mL.At 4×MIC concentration, the peptides inhibited S.mutans biofilm formation by over 60% and suppressed dual-species biofilms of F.nucleatum and P.gingivalis by more than 85%.SEM imaging demonstrated that AMP treatment induced bacterial cell deformation and rupture.Cytotoxicity assays identified YHX-5 and YHX-7 as exhibiting low cytotoxicity.Haemolysis tests further confirmed that YHX-5 possesses low haemolytic activity and high cell selectivity.In conclusion, YHX-5 demonstrates potent antimicrobial and anti-biofilm activity combined with low cytotoxicity and minimal haemolytic risk, establishing it as a promising novel agent for oral antibacterial applications.

参考文献

[1] HAYASHI C, GUDINO C V, GIBSON F C, et al. REVIEW: Pathogen-induced inflammation at sites distant from oral infection: Bacterial persistence and induction of cell-specific innate immune inflammatory pathways: Pathogen-induced inflammation[J].Molecular Oral Microbiology, 2010, 25(5):305-316.
[2] 童跃林, 吴越. 常见口腔问题及其护理进展[J].日用化学工业(中英文), 2023, 53(5): 567-573.
TONG Y L, WU Y. Research progress on the oral problems and their nursing methods[J].China Surfactant Detergent & Cosmetics, 2023, 53(5): 567-573.
[3] BRITO L C N, TELES F R, TELES R P, et al. Use of multiple-displacement amplification and checkerboard DNA-DNA hybridization to examine the microbiota of endodontic infections[J].Journal of Clinical Microbiology, 2007, 45(9):3039-3049.
[4] XU H, HAO W J, ZHOU Q, et al. Plaque bacterial microbiome diversity in children younger than 30 months with or without caries prior to eruption of second primary molars[J].PLoS One, 2014, 9(2): e89269.
[5] HUTTENHOWER C, GEVERS D, KNIGHT R, et al. Structure, function and diversity of the healthy human microbiome[J].Nature, 2012, 486(7402):207-214.
[6] HOARE A, SOTO C, ROJAS-CELIS V, et al. Chronic inflammation as a link between periodontitis and carcinogenesis[J].Mediators of Inflammation, 2019, 2019(1):1029857.
[7] 敖日格乐, 胡和珠拉, 胡伊力格其, 等. 蝎毒素抗菌肽CeAP1的生物信息学分析与生物活性鉴定[J].中国病原生物学杂志, 2024, 19(2): 130-136.
AO R, HU H, HU Y, et al. Functional identification and bioinformatics analysis of antimicrobial peptide Ce AP1 from scorpion venom[J].Journal of Pathogen Biology, 2024, 19(2): 130-136.
[8] 黎观红, 洪智敏, 贾永杰, 等. 抗菌肽的抗菌作用及其机制[J].动物营养学报, 2011, 23(4): 546-555.
LI G H, HONG Z M, JIA Y J, et al. Activities and mechanisms of action of antimicrobial peptides[J].Chinese Journal of Animal Nutrition, 2011, 23(4): 546-555.
[9] 周欣宇, 周春才. 抗菌肽及类抗菌肽的设计、合成及应用[J].化学进展, 2018, 30(7): 913-920.
ZHOU X Y, ZHOU C C. Design, synthesis and applications of antimicrobial peptides and antimicrobial peptide-mimetic copolymers[J].Progress in Chemistry, 2018, 30(7): 913-920.
[10] 李剑勋, 步雨珊, 刘银雪, 等. α-螺旋抗菌肽YH X-1的从头设计及抑菌活性研究[J].食品科学技术学报, 2022, 40(3):98-104; 111.
LI J X, BU Y S, LIU Y X, et al. De novo design and antibacterial activity of α-helical antimicrobial peptide YHX-1[J].Journal of Food Science and Technology, 2022, 40(3):98-104; 111.
[11] 易华西, 李剑勋, 周炜, 等. 一种抗菌肽及其应用: 中国, CN112940082A[P].2021-06-11.
YI H X, LI J X, ZHOU W, et al. An antimicrobial peptide and its application: China, CN112940082A[P].2021-06-11.
[12] 易华西, 李剑勋, 刘同杰, 等. 一种低分子量抗菌肽YHX-2及其组合物和应用: 中国, CN113336827A[P].2021-09-03.
YI H X, LI J X, LIU T J, et al. A low molecular weight antimicrobial peptide YHX-2, its composition and application: China, CN113336827A[P].2021-09-03.
[13] 易华西, 李剑勋, 公丕民, 等. 抗菌肽YHX-3及其组合物和应用: 中国, CN113336828A[P].2021-09-03.
YI H X, LI J X, GONG P M, et al. Antimicrobial peptide YHX-3, its composition and application: China, CN113336828A[P].2021-09-03.
[14] 易华西, 李剑勋, 刘银雪, 等. 抗菌肽YHX-4及其应用: 中国, CN114516900A[P].2022-05-20.
YI H X, LI J X, LIU Y X, et al. Antimicrobial peptide YHX-4 and its application: China, CN114516900A[P].2022-05-20.
[15] 易华西, 李剑勋, 刘银雪, 等. 抗菌肽YHX-5及其应用: 中国, CN114516899A[P].2022-05-20.
YI H X, LI J X, LIU Y X, et al. Antimicrobial peptide YHX-5 and its application: China, CN114516899A[P].2022-05-20.
[16] 易华西, 刘银雪, 周炜, 等. 抗菌肽YHX-7及其应用: 中国, CN115724909A[P].2023-03-03.
YI H X, LIU Y X, ZHOU W, et al. Antimicrobial peptide YHX-7 and its application: China, CN115724909A[P].2023-03-03.
[17] CHAI Q Q, WU Q, LIU T L, et al. Enhanced antibacterial activity of silica nanorattles with ZnO combination nanoparticles against methicillin-resistant Staphylococcus aureus[J].Science Bulletin, 2017, 62(17):1207-1215.
[18] 阿尔曼·阿卜力孜, 吴龙, 玛依奴尔·阿塔吾拉, 等. 基于网络药理学探讨新疆桑白皮防龋机制及其对主要致龋菌作用的研究[J].口腔疾病防治, 2024, 32(2): 89-100.
ABLIZ A, WU L, ATAULA M, et al. Caries prevention mechanism of the Xinjiang Mori Cortex based on network pharmacology and its effect on the main cariogenic bacteria[J].Journal of Dental Prevention & Treatment, 2024, 32(2): 89-100.
[19] 朱苓, 杨明晨, 赖金娥, 等. 抗菌肽F1对酱油腐败菌的抑菌机理探究及其应用[J].中国调味品, 2025, 50(6):34-39; 61.
ZHU L, YANG M C, LAI J E, et al. Study on antibacterial mechanism of antimicrobial peptide F1 against soy sauce spoilage bacteria and its application[J].China Condiment, 2025, 50(6):34-39; 61.
[20] 朱婷婷, 郑炜, 郑霖波, 等. 水产蛋白抗菌肽保鲜剂对草莓的保鲜机理及效果[J].食品研究与开发, 2024, 45(16): 35-40.
ZHU T T, ZHENG W, ZHENG L B, et al. The preservation mechanism and effects of several aquatic protein antimicrobial peptide preservatives on strawberry[J].Food Research and Development, 2024, 45(16): 35-40.
[21] 伊尔凡·努尔买买提, 热依拉·居热提, 苏比努尔·阿尔肯, 等. 鞣花酸对牙龈卟啉单胞菌和具核梭杆菌的体外抑菌效果[J].北京口腔医学, 2023, 31(6): 400-404.
YIERFAN N, REYILA J, SUBINUER A, et al. Antibacterial effect of Ellagic acid on Porphyromonas gingivalis and Fusobacterium nucleatum in vitro[J].Beijing Journal of Stomatology, 2023, 31(6): 400-404.
[22] HANEY E F, HANCOCK R E W. Peptide design for antimicrobial and immunomodulatory applications[J].Peptide Science, 2013, 100(6):572-583.
[23] 苑洁, 叶建方, 刘泳廷, 等. α-鹅膏毒肽对人正常肝细胞MIHA凋亡的影响[J].毒理学杂志, 2025, 39(4):274-278.
YUAN J, YE J F, LIU Y T, et al. The effect of α-amanitin on the apoptosis of normal human liver cells MIHA[J].Journal of Toxicology, 2025, 39(4):274-278.
[24] 朱宁艺. 抗菌肽Mastoparan-C新型类似物的设计、合成、构效关系及其抑制和逆转抗生素耐药性作用研究[D].兰州: 兰州大学, 2021.
ZHU N Y. Design, synthesis, structure-activity relationship of novel analogs of antimicrobial peptide Mastoparan-C and their effects on inhibiting and reversing antibiotic resistance[D].Lanzhou: Lanzhou University, 2021.
[25] 杭柏林, 李杰, 阿卜杜瓦日斯·艾海提, 等. 抗菌肽Loong-3的分子设计及其抑菌活性影响因素分析[J].中国畜牧兽医, 2025, 52(10): 4952-4963.
HANG B L, LI J, ABIEZ A, et al. Molecular design of antimicrobial peptide Loong-3 and analysis of factors influencing its antimicrobial activity[J].China Animal Husbandry & Veterinary Medicine, 2025, 52(10): 4952-4963.
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