血必净注射液干预甲型H1N1流感重症肺炎小鼠的疗效及机制研究

Study on Treatment of Influenza A H1N1 Induced Severe Pneumonia by Xuebijing Injection

目的:观察血必净注射液(血必净)治疗甲型H1N1流感重症肺炎小鼠的疗效,并初步研究其作用机制。方法:C57BL/6小鼠随机分为正常组、模型组、血必净组,正常组小鼠予20μL PBS滴鼻,模型组小鼠2×105TCID50/20μL甲型H1N1流感病毒液滴鼻感染,建立流感重症肺炎小鼠模型。血必净组小鼠2×105TCID50/20μL甲型H1N1流感病毒液滴鼻感染,并于感染前24 h腹腔注射给药,1次/d,连续给药5 d。通过观察、检测不同时间点各组小鼠的存活率、体重、肺病毒载量、肺湿干比、肺组织病理、肺灌洗液细胞因子等结果,评价血必净干预小鼠流感重症肺炎的疗效,初步探讨其作用机制。结果:模型组小鼠存活率为30%,体重下降明显,肺湿干比明显高于正常组(P<0.05),提示严重肺水肿;肺组织病理显示,双肺弥漫性炎性反应细胞浸润,肺泡上皮细胞脱落、坏死,支气管上皮细胞变性,伴有水肿、瘀血、出血,提示流感重症肺炎模型成立。血必净组小鼠存活率为60%,较模型组为优,但无统计学意义(P>0.05),体重下降较模型组轻,肺湿干比较模型组降低,但无统计学意义(P>0.05);肺组织病理示炎症细胞浸润,肺脏水肿,淤血、出血等严重程度均较模型组轻,提示血必净可减轻甲型流感重症肺炎小鼠的肺损伤。模型组与血必净组肺病毒载量无统计学意义(P>0.05),提示血必净并无抑制病毒复制的作用。血必净组可于感染后1 d、3 d降低TNF-α水平(P<0.05),于感染后3 d降低IL-6水平(P<0.05)。结论:血必净对流感重症肺炎小鼠有减轻肺损伤及死亡保护趋势,但作用不显著,这可能与血必净并无直接抗病毒,但能调节感染早期TNF-α、IL-6等细胞因子水平的作用有关。

Objective： To observe the efficacy of Xuebijing Injection on mice with severe pneumonia induced by influenza A virus and to explore the pathogenesis on mice with severe pneumonia. Methods： C57 BL /6 mice were random grouped into three parts including normal group,model group and Xuebijing group. Mice in normal group were given 20 μL PBS by intranasal administration,while the model group 2 × 105TCID50 /20 μL H1N1 virus in the same method. Mice in Xuebijing group were inoculated virus as volume as model group and then given injecta intraperitoneally daily for 5days. Survival rate,weight changes,viral load,acute pulmonary edema（ lung wet-to-dry ratio） and lung histopathology at different time points were measured to evalue the efficacy of Xuebijing injuection on severe influenza and to study its pathogenesis. Results： The survival rate in model group was 30% with notable weight loss. The acute lung edema calculated by wet-to-dry ratio went up clearly compared with that in model group（ P 0. 05）. Lung histopathology of model group indicated the success of severe pneumonia mice model showing that there were diffuse Inflammatory cell infiltration,sheding and necrosis of alveolar epithelial cells,degeneration of bronchial epithelial cells,pulmonary edema and hemorrhage. The survival rate in model group was 60% superior to model group with light weight lose. Wet-to-dry ratio was alleviated in Xuebijing group,but with no statistical differences（ P 0. 05）. The manifestation of lung histopathology in Xuebijing group was as same as model group but the degree of lung injure in this group was eased dramaticlly. There was no difference in viral load between model group and Xuebijing group（ P 0. 05）. It indicated that Xuebijing had no effect on inhibiting the replication of the influenza virus. It could reduce the level of TNF-αat one and 3 days post infection（ P 0. 05） and the level of IL-6 of3 days infection（ P 0. 05）. Conclusion： Xuebijing could alleviate lung injury of severe pneumonia mice and have the tend of protecting those mice. But the protective effect was not prominent which may be related with its no effect on inhibiting virus but its regulation of some early cytokines,such as TNF-αand IL-6.