金屏汤预防免疫力低下小鼠模型呼吸道合胞病毒感染的实验探究

Jinping Decoction Prevents Respiratory Syncytial Virus Infection by Improving Respiratory Immunity

    摘要
  • 摘要:
      目的  探讨金屏汤(JP)对免疫力低下小鼠的免疫调节作用及继发呼吸道合胞病毒(RSV)感染的预防效果。
      方法  将90只Balb/c小鼠随机分为空白对照组，模型组，匹多莫德(0.3 g·kg-1)组，金屏汤高剂量(55.2 g·kg-1)组、中剂量(27.6 g·kg-1)组、低剂量(13.8 g·kg-1)组, 每组15只。通过腹腔注射地塞米松构建免疫力低下小鼠模型, 分别以生理盐水，匹多莫德以及高、中、低剂量金屏汤灌胃7 d后, 滴鼻感染RSV, 空白组滴鼻等体积生理盐水。记录小鼠体质量, 拍照记录脾脏体积, 流式细胞术检测脾脏、肠系膜淋巴结、肺组织中免疫细胞亚群CD4+T、CD8+T、NK、DC比例; qPCR法检测肺部病毒载量，干扰素(Interferon, IFN)-β、干扰素刺激基因(Interferon stimulating gene, ISG)15 mRNA表达水平; HE染色评估肺组织病理学改变; 流式细胞术检测肺泡灌洗液(BALF)中淋巴细胞浸润及肺组织中活化CD8+T细胞(CD8+CD69+T)、记忆性T细胞比例变化。
      结果  金屏汤显著恢复了模型小鼠体质量及肠系膜淋巴结、肺组织中CD8+T、NK、DC细胞的比例(P < 0.05, P < 0.01);qPCR结果提示, 与模型组相比, 金屏汤可降低小鼠肺部RSV载量, 提高IFN-β、ISG15 mRNA的表达(P < 0.05, P < 0.01, P < 0.001);流式细胞术结果显示, 与模型组相比, 金屏汤组小鼠肺组织淋巴细胞浸润显著减少(P < 0.01);HE染色及流式细胞检测结果提示, 与模型组相比, 金屏汤组可改善小鼠肺组织病理学形态, 同时促进CD8+T细胞活化, 增加记忆性T细胞的比例(P < 0.01)。
      结论  金屏汤可提高模型小鼠免疫力, 同时可通过提高Ⅰ型干扰素及CD8+T细胞介导的抗病毒免疫应答, 有效预防RSV感染, 缓解肺部炎症病理损害, 并增强RSV感染后的免疫记忆。

     

    Abstract:
      OBJECTIVE  To investigate the immunoregulatory effect of Jinping decoction (JP) on immunocompromised mice and its preventive effect on RSV infection.
      METHODS  90 Balb/c mice were randomly divided into control group, model group, pidotimod group (0.3 g·kg-1), JP high dose group (55.2 g·kg-1), middle dose group (27.6 g·kg-1), low dose group (13.8 g·kg-1), 15 mice in each group. The immunocompromised mouse model was constructed by intraperitoneal injection of dexamethasone. After 7 days of intragastric administration with normal saline, pidotimod, high, medium low doses of JP, RSV was infected intranasally. The control group received nasal drops of the same volume of normal saline. The body weight of mice was recorded, and the spleen volume was photographed. Flow cytometry was used to detect the proportion of immune cell subsets: CD4+T, CD8+T, NK and DC in spleen, mesenteric lymph nodes and lung tissues. Lung viral load, expression levels of IFN-β mRNA and ISG15 mRNA were detected by qPCR. HE staining was used to evaluate the pathological changes of lung tissue. Lymphocyte infiltration in bronchoalveolar lavage fluid (BALF), the proportion of activated CD8+T cells (CD8+CD69+T) and memory T cells in lung tissue were detected by flow cytometry.
      RESULTS  JP treatment significantly restored the body weight of model mice, and the proportion of CD8+T, NK and DC cells in mesenteric lymph nodes and lung tissue. qPCR results illustrated that compared with the model group, the JP treatment group could reduce the RSV load in the lungs of mice and increase the mRNA expression of IFN-β and ISG15 (P < 0.05, P < 0.01, P < 0.001). Flow cytometry showed that lymphocyte infiltration in lung tissue of mice in the JP treatment group was significantly reduced compared with the model group (P < 0.01). The results of HE staining and flow cytometry revealed that JP could improve the histopathological morphology of lung in mice, promote the activation of CD8+T cells and increase the proportion of central memory T cells (P < 0.01).
      CONCLUSION  JP can improve the immunosuppression of model mice, and effectively prevent RSV infection, alleviate the pathological damage of lung inflammation and enhance the immune memory after RSV infection by increasing the antiviral immune response mediated by type Ⅰ interferon and CD8+T cells.

     

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