目的探讨不同构型人α防御素-5(HD-5)多肽对单纯疱疹Ⅱ型病毒(HSV-2)感染的抑制作用,分析不同构型HD-5抗HSV-2活性的差异。方法 HSV-2病毒感染体外培养的绿猴肾细胞(Vero细胞),分别给予100μg/mL的HD-5/N、HD-5/Acm和HD-5/A肽处理,72h后观察细胞病变效应(CPE),并通过CCK-8法测定细胞保护率;建立HSV-2感染豚鼠生殖器实验动物模型,按照症状评分方法对不同构型HD-5多肽的体内抗HSV-2病毒感染作用进行分析。结果 HD-5/N与HD-5/Acm均能显著抑制HSV-2感染体外培养的Vero细胞(P〈0.01),而HD-5/A的抗病毒作用不明显;阴道内给予HD-5/N多肽12.5μg.kg-1.d-1治疗8d能显著改善HSV-2感染豚鼠的生存状况,提高动物30d存活率(P〈0.01),外阴感染症状评分显著降低(P〈0.01),而HD-5/Acm治疗效果并不明显。结论天然结构HD-5多肽(HD-5/N)在体外或豚鼠阴道内应用均可发挥显著抗HSV-2感染的作用,二硫键缺失会显著影响HD-5多肽的体内抗病毒效果。
Objective To investigate the inhibitive effects of human α defensin 5 polypeptide(HD-5) on herpes simplex virus type 2(HSV-2) infection,and analyze the inequable anti-HSV-2 activity of HD-5 with different configurations.Methods Vero cells infected with HSV-2 in vitro were treated respectively with three different configurations of HD-5,including HD-5/N,HD-5/Acm and HD-5/A at a dose of 100 μg/mL.72 hours later,cytopathic effect(CPE) was observed under microscope,and the cell viability was determined by CCK-8 assay.Female guinea pigs with genitalias infected with HSV-2 were used to analyze the in vitro anti-HSV-2 activity of HD-5 determined by symptom score.Results Both HD-5/N and HD-5/Acm could significantly inhibited the infection of Vero cells by HSV-2 in vitro(P0.01),while the antiviral activity of HD-5/A was not obvious.HD-5/N given through vagina at dose of 12.5 μg·kg·-1·d-1for 8 days could significantly improve the living conditions and survival probability of guinea pigs infected with HSV-2(P0.01),and the symptom score of genital infection was significantly lower(P0.01),compared with saline control group.While there was little therapeutic effect of HD-5/Acm used in vitro for the treatment of HSV-2 infection.Conclusion HD-5 peptide with natural configuration displays a significant anti-HSV-2 ability either in vitro or in vitro.The lack of disulfide bonds significantly affects the antiviral activity of HD-5 in vitro.