地龙提取液对卵清蛋白诱导哮喘小鼠气道重塑及肺组织MMP2、MMP9、TIMP-1表达的影响  

Effects of earthworm extract on airway remodeling and expression of MMP2, MMP9 and TIMP-1 in lung tissues of asthmatic mice induced by ovalbumin

作　　者：张胜碧 刘燕玲 李岚[3] ZHANG Sheng-bi;LIU Yan-ling;LI Lan(Department of Pharmacy,Wenzhou Hospital of Traditional Chinese Medicine,Wenzhou ZHEJIANG 325000,China;Pharmacy,Jinhua Hospital of Traditional Chinese Medicine,Jinhua ZHEJIANG 321000,China;The First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou ZHEJIANG 310000,China)

机构地区：[1]温州市中医院药剂科,浙江温州325000 [2]金华市中医医院药房,浙江金华321000 [3]浙江中医药大学附属第一医院,浙江杭州310000

出　　处：《中国新药与临床杂志》2021年第7期523-530,共8页Chinese Journal of New Drugs and Clinical Remedies

基　　金：浙江省自然科学基金资助项目(LY15H270006)。

摘　　要：目的探讨地龙提取液对哮喘小鼠气道重塑及肺组织基质金属蛋白酶(MMP)2、MMP9、基质金属蛋白酶组织抑制剂(TIMP)-1表达的影响。方法将60只BALB/c小鼠随机均分为正常组,模型组,地龙提取液低、中、高剂量(50、100、200 mg·kg^(-1))组和地塞米松(0.5 mg·kg^(-1))组。除正常组外,各组小鼠采用卵清蛋白(Ova)建立哮喘模型。地龙提取液组每日给药1次,地塞米松组每周给药3次,连续给药8周。Ova末次激发哮喘后24 h,测定小鼠气道反应性;收集支气管肺泡灌洗液(BALF)对炎症细胞进行计数;ELISA法测定BALF中嗜酸性粒细胞趋化因子(Eot)、白细胞介素(IL)-4、IL-5、IL-13含量和血浆Ova特异性IgE水平;通过HE和AB-PAS染色观察肺组织病理学变化;Western blot法检测肺组织中MMP2、MMP9、TIMP-1蛋白表达水平。结果与模型组相比,地龙提取液各剂量组哮喘小鼠的肺阻力降低,肺顺应性提高,BALF中的细胞总数和嗜酸性粒细胞、巨噬细胞、淋巴细胞、中性粒细胞数量减少,Eot、IL-4、IL-5、IL-13含量和血浆Ova特异性IgE水平降低,肺部炎症细胞浸润减轻,肺组织中MMP2、MMP9、TIMP-1蛋白表达下降均呈剂量依赖性趋势,其中地龙提取液高剂量组和地塞米松组无显著差异(P> 0.05)。此外,高剂量地龙提取液和地塞米松能显著降低MMP2/TIMP-1值(P <0.05),但对MMP9/TIMP-1值无显著影响(P> 0.05),且高剂量地龙提取液的作用显著强于地塞米松(P <0.05)。结论地龙提取液对哮喘小鼠气道重塑具有一定的改善作用,其作用机制可能与调节MMPs/TIMP-1平衡有关。AIM To investigate the effects of earthworm extract on airway remodeling and expression of matrix metalloproteinase(MMP)2, MMP9 and tissue inhibitor of metalloproteinase(TIMP)-1 in lung tissues of asthmatic mice induced by ovalbumin(Ova). METHODS Totally 60 BALB/c mice were randomly divided into six groups, including normal group, model group and low, medium and high dose group of earthworm extract(50, 100, 200 mg·kg^(-1)) group as well as dexamethasone(0.5 mg·kg^(-1)) group. Ova was used to establish the asthmatic murine model. The earthworm extract was given once a day and dexamethasone was given three times a week for 8 weeks. Twenty-four hours after the last Ova inhalation, the airway reactivity of mice was determined. Bronchoalveolar lavage fluid(BALF) was collected to count the number of inflammatory cells. The contents of eotaxin, IL-4, IL-5, IL-13 in BALF and the plasma level of Ova-specific IgE were determined by ELISA. HE staining and AB-PAS staining were used for pathological examination of lung tissue. The expressions of MMP2, MMP9, TIMP-1 in lung tissues were detected by Western blot. RESULTS Compared with the model group, in each dose group of earthworm extract, the lung resistance of asthmatic mice was decreased and the lung compliance was increased;the number of total cells and inflammatory cells including eosinophils, macrophages, lymphocytes and neutrophils in BALF decreased, as well as the content of eotaxin, IL-4, IL-5, IL-13 in BALF and plasma level of Ovaspecific IgE;the infiltration of inflammatory cells in lung was alleviated and the expression of MMP2, MMP9 and TIMP-1 in lung tissue declined,with the tendency of dose-dependent. Among them, there was no significant difference between the high-dose earthworm extract group and the dexamethasone group(P > 0.05). Besides, high-dose earthworm extract and dexamethasone could significantly reduce MMP2/TIMP-1(P < 0.05) while had no significant effect on MMP9/TIMP-1(P > 0.05), and the effect of high-dose earthworm extract was significantly strong

关 键 词：地龙 哮喘 气道重塑 基质金属蛋白酶类 基质金属蛋白酶抑制剂 

分 类 号：R969.1[医药卫生—药理学]