抗菌肽JH-3对耐甲氧西林金黄色葡萄球菌的抑菌和抗炎作用研究  

Study on the bactericidal activity of JH-3 against MRSA(Methicilin-resistant Staphyloccocus aureus)and its inhibitory effect on MRSA-induced inflammatory response in A549 cells

作　　者：赵娅娅 刘欣欣 赵雪芹[1] 刘双双 朱春玲 王恒 夏小静[1] 张慧辉[1] 王青[1] 徐彦召[1] 杭柏林[1] 孙亚伟[1] 陈仕钧[1] 胡建和[1] 王磊[1] ZHAO Ya-ya;LIU Xin-xin;ZHAO Xue-qin;LIU Shuang-shuang;ZHU Chun-ling;WANG Heng;XIA Xiao-jing;ZHANG Hui-hui;WANG Qing;XU Yan-zhao;HANG Bo-lin;SUN Ya-wei;CHEN Shi-jun;HU Jian-he;WANG Lei(College of Animal Science and Veterinary Medicine,Henan Institute of Science and Technology,Xinxiang 453003,China)

机构地区：[1]河南科技学院动物科技学院,河南新乡453003

出　　处：《中国预防兽医学报》2020年第7期712-719,共8页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(31702259、31520103917);河南省农业科技攻关项目(182102110347);国家级大学生创新创业训练计划项目(201910467013);国家重点研发计划子课题(2016YFD0500708-04)。

摘　　要：为研究抗菌肽JH-3对耐甲氧西林金黄色葡萄球菌(Methicilin-resistant Staphyloccocus aureus,MRSA)ATCC25923的杀菌和抗炎作用,本研究通过琼脂扩散和最小抑菌浓度(MIC)检测了抗菌肽JH-3对ATCC25923的抑菌作用,结果显示JH-3对ATCC25923具有良好的杀菌作用,1 MIC的JH-3在20 min内可使ATCC25923活菌数降低1000倍;利用结晶紫染色和电镜观察检测JH-3对ATCC25923生物膜形成的影响,结果显示较低浓度的JH-3(0.5 MIC)可以显著降低ATCC25923生物被膜的形成;通过细胞毒性试验和RT-PCR在细胞水平上评价了JH-3对ATCC25923介导的LDH和细胞因子释放的影响,结果显示JH-3(50μg/mL)可以显著降低ATCC25923介导的A549细胞LDH的释放,同时JH-3还具有剂量依赖性的降低ATCC25923诱导的A549细胞炎症相关细胞因子(IL-1β、IL-2和IL-6)的表达;对其他8株不同的MRSA进行抑菌性测定,结果显示JH-3对这些菌株均有很好的杀菌作用,20 min内均可使活菌数降低至103 cfu/mL^107 cfu/mL。本研究首次证实了JH-3能够有效杀灭MRSA及抑制MRSA介导的炎症作用,为新型抗菌药物的研发奠定基础。In order to solve the clinical treatment difficulties brought by the appearance of methicilin-resistant S.aureus(MRSA),the antibacterial and anti-inflammatory effects of antibacterial peptide JH-3 on methicillin-resistant S.aureus ATCC25923 was studied.In this study,the antibacterial effect of antibacterial peptide JH-3 on ATCC25923 was tested by agar diffusion and minimum inhibitory concentration(MIC).The effect of JH-3 on the biofilm formation of ATCC25923 was examined by crystal violet staining and electron microscopy.The effects of JH-3 on ATCC25923 mediated LDH and cytokine release were examined at the cellular level by cytotoxicity tests and RT-PCR.Finally,the broad spectrum of JH-3 bactericidal activity was evaluated.The results showed that JH-3 had a good bactericidal effect on ATCC25923,and 1 MIC of JH-3 could reduce the viable number of ATCC25923 by 1000 times within 20 min.Lower concentration of JH-3(0.5 MIC)can significantly reduce the formation of ATCC25923 biofilm.In addition,MRSAATCC25923-mediated LDH release in A549 cells was largely reduced by JH-3(50μg/mL),meanwhile,inflammatory cytokines(IL-1β,IL-2 and IL-6)in MRSAATCC25923 infected A549 cells was also decreased by JH-3 in a dose-dependent manner.Eight MRSA strains were found to be efficiently killed by JH-3,the viable count decreased from 103 cfu/mL to 107 cfu/mL within 20 min.This study proved for the first time that JH-3 can effectively kill MRSA and inhibit MRSA mediated inflammation,thus laying a foundation for the development of new antibacterial drugs.

关 键 词：抗菌肽 JH-3 耐甲氧西林金黄色葡萄球菌 杀菌作用 炎症反应 

分 类 号：S852.4[农业科学—基础兽医学]