家蝇抗菌肽MAF-1A体外抗甲型流感病毒活性及机制研究

Effects and mechanisms of antimicrobial peptide MAF-1A from Musca domestica anti-influenza A virus in vitro

目的探讨家蝇抗菌肽MAF-1A体外抗甲型流感病毒(IAV)活性及其潜在机制。方法通过观察CPE、MTT法及qRT-PCR评价MAF-1A体外抗IAV活性,采用MTT法测定MAF-1A对MDCK细胞的毒性;利用透射电镜(TEM)技术、血凝抑制试验和神经氨酸酶抑制试验进一步分析MAF-1A抗IAV的作用机制。结果MAF-1A对IAV的半数有效浓度(EC50)为(89.8±2.97)μg/mL,而对MDCK细胞的毒性较小;MAF-1A可直接破坏IAV形态结构的完整性;浓度为1.56μg/mL的MAF-1A即可抑制IAV引起的红细胞凝集;对神经氨酸酶具有抑制作用,IC50为(134.7±10.31)μg/mL。结论抗菌肽MAF-1A具有体外抗IAV活性,除能直接破坏IAV的结构外,还可能通过与血凝素HA1亚基结合、抑制神经氨酸酶的活性而阻止IAV的感染,提示MAF-1A具有多靶点抗IAV的作用。

The purpose of our research was to explore the effects and potential mechanisms of antimicrobial peptide MAF-1A from Musca domestica on anti-influenza A virus (IAV) activity.In this research,activity of MAF-1A against IAV in vitro were detected with the CPE,MTT assay and quantitative real-time PCR.Cytotoxicity of MAF-1A on MDCK cells was determined by MTT assay.Transmission electron microscope (TEM),hemagglutination inhibition assay and neuraminidase inhibition assay were used to investigate the mechanism for its anti-IAV activities.MAF-1A could significantly inhibit the infection of IAV in vitro,and its median effective concentration (EC50) was (89.8±2.97)μg/mL,and the low cytotoxicity showed on MDCK cells.The results of TEM indicated that MAF-1A had an obvious destructive effect on IAV.The hemagglutination induced by IAV was inhibited by MAF-1A concentration of 1.56μg/mL.MAF-1A had the inhibition on NA (IC50 (134.7±10.31)μg/mL).This study suggests that MAF-1A have remarkably inhibitory effects on IAV,the mechanisms may be associated with the virion damage,combination with hemagglutinin (HA) HA1 subunit and neuraminidase inhibition,which suggest that the anti-IAV of MAF-1A may be through multi-targets.