不同构型人α防御素-5多肽抗HSV-2感染的实验研究

摘要

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100 μg/mL的HD-5/N、HD-5/Acm和HD-5/A肽处理,72 h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12.5 μg·kg-1·d-1治疗8 d能显著改善HSV-2感染豚鼠的生存状况,提高动物30 d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 a 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 in fection of Vero cells by HSV 2 in vitro(P<0. 01) ,while the antiviral activity of HD 5/A was not obvious. HD 5/N given through vagina at dose of 12. 5 μg o kg o -1 o d"1 for 8 days could significantly improve the living conditions and survival probability of guinea pigs infected with HSV 2(P<0. 01) , and the symptom score of genital infection was significantly lower(P-<0. 01) , com pared with saline control group. While there was little therapeutic effect of HD 5/Acm used in vitro for the treatment of HSV 2 in fection. 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.