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目的探讨人α防御素-5(humanα-defensin-5,HD-5)多肽体外抗单纯疱疹Ⅱ型病毒(herpes simplex virustype2,HSV-2)的作用及其机制。方法以绿猴肾细胞(Vero)为宿主细胞,采用CCK-8法首先分析10~200μg/ml浓度范围内HD-5多肽的细胞毒性,再以HSV-2感染Vero细胞,分别于感染前后和感染的同时加入100μg/ml的HD-5多肽,72h后测定细胞保护率,结合细胞病变效应观察,分析HD-5抗HSV-2的作用效果和起效环节,并通过改变培养基中血清的浓度,观察血清对HD-5抗病毒作用的影响。结果在100μg/ml浓度以内,HD-5多肽几乎无细胞毒性,提前或同时加入HD-5多肽可以显著抑制HSV-2病毒引起的细胞病变反应、提高细胞保护率(P<0.01),而在HSV-2感染2h后加入HD-5多肽对Vero细胞的保护作用不明显(P>0.05);血清浓度会显著影响HD-5多肽的抗HSV-2活性,血清浓度越高,HD-5多肽对细胞的保护率越低。结论HD-5多肽在体外具有显著的抗HSV-2感染活性,其作用机制主要是通过干扰病毒对细胞的粘附、侵入而发挥作用。
Objective To investigate the effect of human α-defensin-5 (HD-5) polypeptide against herpes simplex virustype2 (HSV-2) in vitro and its mechanism. Methods Vero cells were used as host cells. The cytotoxicity of HD-5 peptides was detected by CCK-8 method. The Vero cells were infected with HSV-2, HD-5 polypeptide was added simultaneously with 100μg / ml of infection, and the cell protection rate was measured after 72 hours. Combined with cytopathic effect observation, the effect and the effect of HD-5 anti-HSV-2 were analyzed. Concentration, observe the effect of serum on HD-5 antiviral effect. Results HD-5 polypeptide had almost no cytotoxicity at the concentration of 100μg / ml. The addition of HD-5 polypeptide in advance or at the same time significantly inhibited the cytopathic effect induced by HSV-2 virus and increased the cell protection rate (P <0.01) HSV-2 infection 2 h after the addition of HD-5 polypeptide on Vero cells showed no significant protective effect (P> 0.05); serum concentrations of HD-5 polypeptide will significantly affect the anti-HSV-2 activity, the higher the serum concentration of HD-5 polypeptide The lower the protection rate of cells. Conclusion The HD-5 polypeptide has a significant anti-HSV-2 activity in vitro. The mechanism of action of HD-5 polypeptide is mainly through interfering with cell adhesion and invasion.