王健英　陈晓俊　张磊　王凯　侯剑伟　符胜光　潘颖宜　袁颖

摘要：目的 觀察杜仲皮、杜仲雄花醇提物对鸡卵清蛋白（OVA）所致小鼠气道变应性炎症的影响，探讨其作用机制。方法 实验第0、7日，OVA腹腔注射致敏，继而滴鼻激发21 d，建立小鼠气道变应性炎症模型。实验小鼠随机分为正常组、模型组、杜仲皮醇提物组（杜仲皮组）、杜仲雄花醇提物组（杜仲雄花组）和地塞米松组，各给药组给予相应药物灌胃。肺组织行HE、PAS染色，观察支气管及周围肺组织病理改变，ELISA检测小鼠血清OVA-IgE、白细胞介素（IL）-4、干扰素-γ（IFN-γ）、IL-13水平，免疫组化检测小鼠肺组织细胞间黏附分子-1（ICAM-1）、血管内皮生长因子（VEGF）、基质金属蛋白酶9（MMP9）、组织金属蛋白酶抑制剂1（TIMP1）表达，流式细胞术检测小鼠外周血Th17+细胞表达，RT-PCR检测小鼠肺组织肿瘤坏死因子-α（TNF-α）、IL-6 mRNA表达。结果 模型组小鼠肺组织可见嗜酸性粒细胞及其他炎症细胞浸润，支气管痉挛，黏液分泌增加，杜仲皮组和杜仲雄花组小鼠肺组织病理明显改善。与正常组比较，模型组小鼠血清OVA-IgE、IL-4、IL-13水平升高，IFN-γ水平降低（P<0.05，P<0.01），肺组织ICAM-1、VEGF、MMP9表达增强，TIMP1表达降低（P<0.01），外周血IL-17+细胞增加，肺组织TNF-α、IL-6 mRNA表达均升高（P<0.01）；与模型组比较，杜仲皮组和杜仲雄花组小鼠血清OVA-IgE、IL-4、IL-13水平明显下调（P<0.01），ICAM-1、VEGF表达明显下调（P<0.05，P<0.01），TIMP1表达升高，外周血IL-17+细胞降低，肺组织IL-6 mRNA表达降低（P<0.05，P<0.01），杜仲皮组还可诱导IFN-γ分泌（P<0.01），下调TNF-α mRNA表达（P<0.05），杜仲雄花组MMP9表达明显下调（P<0.05）。结论 杜仲皮、杜仲雄花醇提物可能通过降低模型小鼠OVA-IgE产生，抑制Th2类细胞因子分泌，下调Th17细胞，抑制促炎细胞因子表达，对气道变应性炎症有一定抑制作用。

关键词：杜仲皮；杜仲雄花；气道变应性炎症；细胞因子；黏附分子；小鼠

DOI：10.3969/j.issn.1005-5304.2018.03.010

中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2018）03-0042-06

Abstract： Objective To investigate the effects of alcohol extract of bark and male flower of Eucommia ulmoides Oliv. on airway allergic inflammation induced by chicken ovalbumin （OVA） in mice； To explore its mechanism of action. Methods On day 0， day 7， mice were intraperitoneally injected OVA for sensitization， followed by nasal stimulation for 21 days to establish airway allergic inflammation mice models. The mice were divided into normal group， model group， alcohol extract of bark of Eucommia ulmoides Oliv. group， alcohol extract of male flower of Eucommia ulmoides Oliv. group， and Dexamethasone group. Each medication group was given relevant medicine for gavage. The lung tissue was embedded in HE and PAS dyeing， to observe the pathological changes of bronchus and

surrounding lung. The levels of serum OVA-IgE， IL-4， IFN-γ and IL-13 were measured by ELISA. The expression of ICAM-1， VEGF， MMP9 and TIMP1 were detected by immunohistochemistry. Flow cytometry was used to detect the expression of Th17 cells in peripheral blood. The expressions of TNF-α and IL-6 mRNA in lung tissue were detected by RT-PCR. Results The model group showed changes of airway allergic inflammatory such as eosinophils and other inflammatory cell infiltration， bronchial spasm， and mucus secretion. Lung histopathology in alcohol extract of bark and male flower of Eucommia ulmoides Oliv. groups was improved significantly （P<0.05）. Compared with the normal group， the levels of serum OVA-IgE， IL-4 and IL-13 increased in model group， while the level of IFN-γ decreased （P<0.05， P<0.01）. The expressions of ICAM-1， VEGF and MMP9 increased， while the expression of TIMP1 decreased （P<0.01）； peripheral blood IL-17+cells increased （P<0.01）； the expressions of TNF-α and IL-6 mRNA increased. Compared with the model group， the levels of serum OVA-IgE， IL-4 and IL-13 decreased in alcohol extract of bark and male flower of Eucommia ulmoides Oliv. groups （P<0.05， P<0.01）； the expressions of ICAM-1 and VEGF decreased （P<0.05， P<0.01）； the expression of TIMP1 increased. Alcohol extract of bark and male flower of Eucommia ulmoides Oliv. could down-regulate IL-17+cells， reduce the expression of IL-6 mRNA （P<0.05， P<0.01）. Alcohol extract of bark of Eucommia ulmoides Oliv. group could induce the secretion of IFN-γ （P<0.01）， and down-regulate the expression of TNF-α mRNA （P<0.05）. Alcohol extract of male flower of Eucommia ulmoides Oliv. group could significantly down-regulate the expression of MMP9 （P<0.05）. Conclusion Alcohol extract of bark and male flower of Eucommia ulmoides Oliv. can induce the production of OVA-IgE， inhibit secretion of Th2 cytokines， inhibit the expression of adhesion molecules， depress Th17 cells， so as to inhibit the airway allergic inflammation.

Keywords： bark of Eucommia ulmoides Oliv.； male flower of Eucommia ulmoides Oliv.； airway allergic inflammation； cytokines； adhesion molecules； mice

杜仲Eucommia ulmoides Oliv.属杜仲科植物，是我国特有植物，杜仲传统药用部位为树皮，具有补肝肾、强筋骨的功效，用于肝肾不足之腰膝酸痛、筋骨无力。杜仲雄花目前多作为花茶应用，有研究显示，其具有良好的镇静催眠及抗氧化、抑菌作用[1-2]。前期研究表明，杜仲皮、杜仲雄花均有一定的抗炎、调节免疫、镇痛、抑菌等作用[3]。……