海南红树林淡紫拟青霉胞外多糖抗HSV-1作用研究

Paecilomyces Lilacinus Isolated from Mangrove in Hainan Province

目的探讨红树林淡紫拟青霉胞外多糖体内抗单纯疱疹病毒-1(HSV-1)作用。方法选取昆明种小鼠75只,随机分为正常对照组,病毒对照组,低、中、高剂量组,每组15只。正常对照组给予DMEM 0.5 ml腹腔注射,其余4组给予HSV-10.5 ml腹腔注射。次日低、中、高剂量组小鼠分别经腹腔注射淡紫拟青霉胞外多糖6、8、10 g/(kg·d);正常对照组和病毒对照组腹腔注射生理盐水0.5 ml,连续注射7 d。观察并比较各组小鼠肝脏病理变化;5组小鼠脑组织匀浆上清液接种于Vero细胞,观察其细胞病变情况;提取同一部位脑组织病毒基因组DNA,检测病毒DNA相对含量及血清干扰素-γ(IFN-γ)、白介素-12(IL-12)和肿瘤坏死因子-α(TNF-α)含量。结果与正常对照组比较,病毒对照组和低剂量组小鼠肝脏汇管区大量炎性细胞浸润,部分肝细胞肿胀;中剂量和高剂量组炎性细胞浸润明显减少。与病毒对照组相比,中、高剂量组脑组织HSV-1病毒DNA相对含量减低(P<0.01),低剂量组病毒DNA相对含量与病毒对照组比较差异无统计学意义(P>0.05)。与正常对照组比较,其余4组小鼠血清IFN-γ、IL-12、TNF-α水平均显著升高(P<0.01);高剂量组IFN-γ水平高于病毒对照组和低、中剂量组(P<0.01);中、高剂量组IL-12水平高于病毒对照组和低剂量组(P<0.01)。与病毒对照组比较,低、中、高剂量组TNF-α水平均显著降低,且中、高剂量组低于低剂量组(P<0.01)。结论红树林淡紫拟青霉胞外多糖具有一定抗HSV-1作用,可能是通过抑制病毒复制、刺激IFN-γ和IL-12分泌,抑制TNF-α产生完成的。

Objective To observe the anti-HSV-1 activity of extracellular polysaccharides(EPS)from Paecilomyces lilacinus isolated from Mangrove in Hainan mangrove.Methods Seventy-five Kunming mice were randomly divided into normal control group,virus control group,and low-,medium-and high-dose EPS groups,with 15 mice in each group.DMEM 0.5 ml was injected intraperitoneally in the normal control group and HSV-10.5 ml was injected intraperitoneally in the other four groups.On the next day,mice in the low-,medium-and high-dose EPS groups were injected with 6,8 and 10 g/(kg·d)Paecilomyces lilacinus EPS respectively,and mice in the normal control group and the virus control group were injected with 0.5 ml normal saline for 7 consecutive days.The pathological changes of liver in each group were observed and compared.Vero cells were inoculated in the supernatant of brain homogenate of five groups to observe the cytopathy.The viral genomic DNA was extracted from the same part of brain tissue to detect the relative content of viral DNA and the content of interferon-γ(IFN-γ),interleukin-12(IL-12)and tumor necrosis factor-α(TNF-α)in serum.Results Compared with the normal control group,a large number of inflammatory cells were infiltrated in the portal area of mice in the virus control group and low-dose EPS group,and some hepatocytes were swollen.Inflammatory cells infiltrated in the medium-dose and high-dose EPS groups were significantly reduced.Compared with the virus control group,the relative content of HSV-1 DNA in the brain tissue of the medium and high-dose EPS groups was decreased(P<0.01),and there was no significant difference in the relative content between the low-dose EPS group and the virus control group(P>0.05).Compared with those in normal control group,the levels of IFN-γ,IL-12 and TNF-αin the other four groups were significantly higher(P<0.01).The levels of IFN-γin the high-dose EPS groups were higher than those in the virus control group and the low-and medium-dose EPS groups(P<0.01).The levels of IL-12 in the medium-and high-dose EPS groups were higher than those in the virus control group and the low-dose EPS group(P<0.01).Compared with that in virus control group,the TNF-αlevels in the low,medium and high-dose EPS groups were significantly lower,while the levels in the medium and high-dose EPS groups were lower than that in the low-dose EPS group(P<0.01).Conclusion The EPS may play an anti-HSV-1 role by inhibiting the virus replication and stimulating secretion of IFN-γand IL-12,and prevent the secretion of TNF-α.