小鼠单纯疱疹病毒性面瘫的预防和复发模型的建立

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目的建立1型单纯疱疹病毒(herpes simplex virus type 1,HSV-1)潜伏感染再激活导致小鼠面瘫的模型,观察免疫球蛋白(IgG)和干扰素对单纯疱疹病毒性小鼠面瘫的预防效果。方法64只4周龄雌性 Balb/c 小鼠,用26G 针头搔刮小鼠耳廓背面近耳根部皮肤,左耳接种 HSV-1 25μl,总剂量1.7×10~7空斑形成单位(plaque forming unit,PFU)/ml,右耳接种25μl磷酸盐缓冲液作为对照,制作小鼠面瘫潜伏模型。按数字表法将小鼠随机分为3组:第一组20只,每天腹腔注射 IgG1 ml/kg,连续注射3 d,待出现面瘫后,面瘫小鼠继续注射3 d;第二组20只,腹腔注射干扰素1.5×10~7IU/kg,连续3 d,待出现面瘫后,面瘫小鼠继续注射3 d;第三组24只,腹腔注射相应量的生理盐水作为对照。观察小鼠面瘫情况,8周后取面瘫恢复的小鼠,腹腔注射环孢素50 mg/kg,建立面瘫复发模型。处死小鼠,分离双侧面神经和三叉神经节,多聚酶链反应(polymerase chain reaction,PCR)检测HSV-1 DNA 的表达。结果第一组小鼠面瘫率50%,面瘫持续时间(7.2±2.2)d(±s,下同);第二组小鼠面瘫率30%,面瘫持续时间(4.5±1.8)d;第三组小鼠面瘫率67%,面瘫持续时间(8.9±2.6)d。经统计学分析,IgG 不能有效降低面瘫发生率及缩短面瘫持续时间(P 值均>0.05),而干扰素可以有效降低面瘫发生率并缩短面瘫持续时间(P 值均<0.05)。注射环孢素后3只面瘫恢复小鼠(3/28)再次出现面瘫,复发率11%。所有面瘫复发小鼠均检测到 HSV-1 DNA,而未复发小鼠均未检测出HSV-1 DNA。结论 HSV-1潜伏感染再激活可能是单纯疱疹病毒性小鼠面瘫复发的原因之一,潜伏病毒的再激活与免疫力低下有关。干扰素可以有效降低单纯疱疹病毒性面瘫发生率和缩短面瘫持续时间;免疫球蛋白(IgG)不能有效降低面瘫发生率和缩短面瘫持续时间。 Objective To establish a model of facial paralysis induced by latent infection of herpes simplex virus type 1 (HSV-1) in mice and to observe the preventive effect of immunoglobulin (IgG) and interferon on the facial paralysis of herpes simplex virus . Methods Sixty-four female Balb / c mice of 4 weeks old were scalded with 26G needles and the left ear was inoculated with 25μl of HSV-1 and the total dose of 1.7 × 10-7 plaque forming unit, PFU) / ml, the right ear was inoculated with 25μl phosphate buffer as a control to make a mouse facial paralysis latent model. The mice were randomly divided into 3 groups according to the digital table method: the first group of 20 mice were injected intraperitoneally with IgG1 ml / kg for 3 days. After facial paralysis, facial paralysis mice were injected for 3 days. The second group of 20 mice , Intraperitoneal injection of interferon 1.5 × 10 ~ 7IU / kg, for 3 days, to be facial paralysis, facial paralysis mice were injected for 3 days; the third group of 24, the corresponding amount of intraperitoneal injection of saline as a control. Mice with facial paralysis were observed. Eight weeks later, mice with facial paralysis recovered were injected intraperitoneally with cyclosporine 50 mg / kg to establish a model of facial paralysis relapse. Mice were sacrificed, the bilateral facial nerve and trigeminal ganglion were isolated, and the expression of HSV-1 DNA was detected by polymerase chain reaction (PCR). The first group of mice with facial paralysis rate of 50%, duration of facial paralysis (7.2 ± 2.2) d (± s, the same below); the second group of mice facial paralysis rate of 30%, facial paralysis duration (4.5 ± 1.8) d; third Group mice facial paralysis rate of 67%, facial paralysis duration (8.9 ± 2.6) d. Statistically, IgG can not effectively reduce the incidence of facial paralysis and shorten the duration of facial paralysis (P> 0.05), while interferon can effectively reduce the incidence of facial paralysis and shorten the duration of facial paralysis (P <0.05). Three facial paralysis recovery mice (3/28) showed facial paralysis again after injection of cyclosporine, the recurrence rate was 11%. HSV-1 DNA was detected in all facial paralysis relapse mice, while no HSV-1 DNA was detected in the non-relapsed mice. Conclusion Reactivation of latent infection with HSV-1 may be one of the reasons for the recurrence of facial paralysis in herpes simplex virus-prone mice. The reactivation of latent virus is related to the low immunity. Interferon can effectively reduce the incidence of herpes simplex virus facial paralysis and shorten the duration of facial paralysis; immunoglobulin (IgG) can not effectively reduce the incidence of facial paralysis and shorten the duration of facial paralysis.
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