犀角地黄汤合银翘散对流感病毒感染的小鼠肺组织及大鼠肺微血管内皮细胞ICAM-1和VCAM-1表达的影响

 目的：研究中医经典合方犀角地黄汤合银翘散(XDY)对流感病毒性肺炎小鼠肺组织及对流感病毒感染的大鼠肺微血管内皮细胞(RPMVECs)中细胞间黏附分子1（ICAM-1）和血管细胞黏附分子1（VCAM-1）表达的影响，探讨其治疗病毒性肺炎的机制。方法：54只雄性BALB/c小鼠随机分为正常组、模型组和XDY组，每组18只，后2组以流感病毒滴鼻感染，XDY组灌胃给予XDY；在感染后的2、4和6 d分别取材，免疫组化法观察肺组织中ICAM-1和VCAM-1表达。从雄性Wistar大鼠中分离并原代培养RPMVECs，设置正常组、病毒组、病毒+XDY含药血清组、肿瘤坏死因子α（TNF-α）组和TNF-α+XDY含药血清组；刺激因素作用24 h后，real-time PCR检测ICAM-1和VCAM-1 mRNA水平，流式细胞术检测ICAM-1和VCAM-1蛋白表达。结果：与正常组比较，模型组肺组织中ICAM-1和VCAM-1表达持续增多(P<0.01)，而XDY组的表达均低于模型组 (P<0.01)；与正常组比较，流感病毒和TNF-α作用的RPMVECs中 ICAM-1和VCAM-1 mRNA及蛋白表达明显升高(P<0.01)，XDY能下调ICAM-1和VCAM-1 mRNA及蛋白表达 (P<0.01)。结论：抑制流感病毒感染导致的RPMVECs黏附分子的表达从而抑制机体的炎症级联反应可能是XDY治疗流感病毒性肺炎的作用机制之一。

Effects of Xijiao Dihuang and Yinqiao San decoction on ICAM-1 and VCAM-1 expression in mouse lung tissues and rat pulmonary microvascular endothelial cells infected with influenza virus

AIM：To investigate the effects of Xijiao Dihuang and Yinqiao San decoction (XDY) on the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in mouse lung tissues and rat pulmonary microvascular endothelial cells (RPMVECs) infected with influenza virus, and to explore its mechanism for treatment of viral pneumonia. METHODS：Fifty-four male BALB/c mice were randomly divided into normal group, model group and XDY group (n=18 in each group). The viral pneumonia model was established by intranasally dripping influenza A (H1N1) virus into the mice. The mice in XDY group were treated with XDY 1 h after dripping the virus. The expression of ICAM-1 and VCAM-1 in lung tissues was examined by immunohistochemical staining 2, 4 and 6 d after infection. On the other hand, RPMVECs were obtained from male Wistar rats and primarily cultured. The cells were randomly divided into control group, virus group, virus+XDY group, tumor necrosis factor α (TNF-α) group and TNF-α+XDY group. The mRNA and protein expression of ICAM-1 and VCAM-1 was evaluated by real-time PCR and flow cytometry 24 h after infection. RESULTS：Virus exposure increased ICAM-1 and VCAM-1 expression in mouse lung tissues (P<0.01), and XDY treatment attenuated this effect (P<0.01). Virus and TNF-α both led to the increases in mRNA and protein expression of ICAM-1 and VCAM-1 in RPMVECs (P<0.01), which were also reduced by treatment with XDY (P<0.01). CONCLUSION：Treatment with XDY decreases virus-induced ICAM-1 and VCAM-1 expression, suggesting an important role of XDY in treatment of viral pneumonia.