海参抗菌肽的活性筛选与功效评价  

Screening and efficacy evaluation of antibacterial peptides from Holothuroidea

作　　者：袁华标 黄靖彤 万鹏[1,3,4] 蔡冰娜 潘剑宇[1,3,4] 张煜航 凌娟[1,3,4] 陈华 YUAN Huabiao;HUANG Jingtong;WAN Peng;CAI Bingna;PAN Jianyu;ZHANG Yuhang;LING Juan;CHEN Hua(CAS Key Laboratory of Tropical Marine Bio-resources and Ecology,South China Sea Institute of Oceanology,Chinese Academy of Sciences,Guangzhou 510301,China;University of Chinese Academy of Sciences,Beijing 100049,China;Southern Marine Science and Engineering Guangdong Laboratory(Guangzhou),Guangzhou 511458,China;Institution of South China Sea Ecology and Environmental Engineering,Chinese Academy of Sciences,Guangzhou 510301,China)

机构地区：[1]中国科学院南海海洋研究所,中国科学院热带海洋生物资源与生态重点实验室,广东广州510301 [2]中国科学院大学,北京100049 [3]南方海洋科学与工程广东省实验室(广州),广东广州511458 [4]中国科学院南海生态环境工程创新研究院,广东广州510301

出　　处：《热带海洋学报》2023年第4期184-194,共11页Journal of Tropical Oceanography

基　　金：广东省海洋经济发展专项(粤自然资合[2022]036、粤自然资合[2020]036、粤自然资合[2020]041);广东省自然科学基金项目(2022A1515010767);广东省重点领域研发计划(2020B1111030004);中国科学院南海生态环境工程创新研究院自主部署项目(ISEE2021PY05);海南省重点研发计划(ZDYF2021SHFZ109)。

摘　　要：本文将已有文献报道的海洋动物源抗菌肽(毒素除外)与海参蛋白组进行比对,结合生物信息学软件,设计、筛选海参抗菌肽,并对其理化特性、抗菌活性与机制、生物相容性与细胞活性等进行研究。研究结果发现,通过软件模拟与抗菌活性预测,最终筛选并固相合成7条海参抗菌肽(H1-H7)。其中,H4(RVHRFLRR)可通过细菌膜电位去极化、膜透化等方式,抑制金黄色葡萄球菌(Staphylococcus aureus)、铜绿假单胞菌(Pseudomonas aeruginosa)、创伤弧菌(Vibrio vulnificus)生长,在较低浓度(31.25μg·mL^(-1))时,可使S.aureus细菌总数降至84.93%±4.21%(p<0.01),P.aeruginosa细菌总数降至95.92%±0.52%(p<0.01),V.vulnificus细菌总数降至77.14%±1.37%(p<0.01)。H4生物相容性好,在<1000μg·mL^(-1)时均未表现出溶血性或细胞毒性,其可能通过与细胞膜表面受体(EGFR、VEGFR2、FGFR1等)发生相互作用进而促进细胞迁移。在浓度为62.50μg·mL^(-1)和250μg·mL^(-1)时,H4均可显著促进L929细胞发生迁移(p<0.05)。实验结果表明,海参抗菌肽H4生物相容性好,兼有抗菌、促愈合功效,可望作为一种潜在功能物质用于感染创面的修复与再生。In this paper,we compared the existing marine animal origin antibacterial peptides reported(Toxin except)with the holothurian proteome,combined with bioinformatics software to design,screen holothurian antibacterial peptides,and to investigate their physicochemical properties,antibacterial activity,mechanism,biocompatibility and cellular activity.It was found that through software simulation and antibacterial activity prediction,7 holothurian bacteriostatic peptides(H1~H7)were finally screened and solid-phase synthesized.Among them,H4(RVHRFLRR)could inhibit the growth of Staphylococcus aureus,Pseudomonas aeruginosa,Vibrio vulnificus through bacterial membrane potential depolarisation or membrane permeabilisation.At a lower concentration(31.25µg·mL^(−1)),it reduced the total bacterial count of S.aureus to 84.93%±4.21%(p<0.01),P.aeruginosa to 95.92%±0.52%(p<0.01)and V.vulnificus to 77.14%±1.37%(p<0.01).It was also found that H4 was biocompatible and did not exhibit haemolytic or cytotoxic properties at<1000µg·mL^(−1).In addition,H4 may interact with cell membrane receptors(EGFR,VEGFR2,FGFR1,etc.)to promote cell migration.H4 significantly promoted the migration of L929 cells at both 62.50µg·mL^(−1) and 250µg·mL^(−1)(p<0.05).The above experimental results indicate that holothurian antibacterial peptide H4 is biocompatible,has both antibacterial and pro-healing effects,and can be used as a potential functional active substance for the repair and regeneration of infected wounds.

关 键 词：海参 抗菌肽 生物信息学 细胞迁移 创伤修复 

分 类 号：P745.3[天文地球—海洋生物学]