宁夏沙枣花乙醇提取物对哮喘模型小鼠气道炎症的实验研究
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摘要
目的:观察沙枣花乙醇提取物对哮喘模型小鼠气道炎症及血清白介素4(Interleukin-4,IL-4)、IgE-免疫球蛋白(IgE-immunoglobulin,IgE)、γ干扰素(Interferon-γ,IFN-γ)含量的影响,探讨其抑制哮喘气道炎症的作用机制。方法:将BALB/c小鼠随机分为6组,分别为正常组、模型组、地塞米松10mg/kg组和沙枣花乙醇提取物4g/kg、8g/kg、16g/kg组,除空白对照组外,其余各组用改良的卵白蛋白多点注射致敏与雾化吸入激发法建立小鼠哮喘模型,连续14 d。小鼠末次激发24 h后,处死各组小鼠留取标本。镜下观察各组哮喘小鼠肺组织病理形态变化及支气管肺泡灌洗液中白细胞计数和分类计数,并进行炎症评分;采用双抗体夹心酶联免疫吸附试验(ELISA)法测定小鼠支气管肺泡灌洗液及血清中IL-4、IgE、IFN-γ含量。结果:与模型组比较,沙枣花乙醇提取物8g/kg、16g/kg组和地塞米松组可显著降低哮喘模型小鼠支气管肺泡灌洗液及血清中IgE、IL-4含量和IL-4/IFN-γ比值;16g/kg、8g/kg组和地塞米松组可显著升高支气管肺泡灌洗液及血清中IFN-γ含量。与模型组比较,沙枣花乙醇提取物16g/kg、8g/kg组和地塞米松组可显著降低白细胞总数、嗜酸性粒细胞及巨噬细胞比率;各给药组可显著升高中性粒细胞比率。病理切片染色结果显示沙枣花乙醇提取物16g/kg、8g/kg组能够明显减轻肺组织细胞炎性浸润等病理症状,与模型组比较,可显著降低炎症评分。结论:沙枣花提取物可通过降低IL-4及IgE含量、升高IFN-γ的含量,调节Th1/Th2免疫平衡而改善哮喘模型小鼠气道炎症。
Objective: To observe the effect of the alcohol extract of Elaeagnus angustifolia blossoms on the respiratory tract inflammation and the serum levels of IL-4,IgE and IFN-γ in the asthmatic mice model,also to explore the mechanism of its inhibition of asthma. Methods: BALB/c mice were randomly divided into 6 groups as followings: the normal group( normal saline),the model group,dexamethasone( 10 mg/kg/d) group and three doses of alcohol extract of Elaeagnus angustifolia blossoms groups( 16 g/kg/d,8 g/kg/d and 4 g/kg/d). Except for the normal group,the asthma models were established in all other groups by improved ovalbumin multipoint injection induced sensitization and atomization inhalation method for 14 d. All mice were sacrificed 24 hours after final stimulation. The pathological changes of lung tissues,counts and classification of white blood cells in bronchoalveolar lavage fluid were observed under microscope,and the inflammatory score was scored. The levels of IL-4,IgE and IFN-γ in bronchoalveolar lavage fluid and serum of mice were determined by double antibody sandwich ELISA. Results: Compared with the model group,the levels of IgE,IL-4 and IL 4/IFN γratio in serum and lavage were significantly decreased in 8 g/kg,16 g/kg/d groups of Elaeagnus angustifolia Blossoms alcohol extract and dexamethasone group in bronchial alveolar asthma model perfusion,the content of IFNγwas significantly increased in 16 g/kg,8 g/kg groups and dexamethasone group in the bronchoalveolar lavage fluid and serum. Compared with the model group,total numbers of white blood cells,eosinophils and mononuclear cell ratio were significantly reduced in Elaeagnus angustifolia blossoms alcohol extract 16 g/kg、8 g/kg group and dexamethasone group,and neutrophil ratio was significantly increased in all alcohol extract groups. Pathological staining showed that the 16 g/kg and 8 g/kg of Elaeagnus angustifolia blossoms alcohol extract significantly reduced the inflammatory infiltration of lung tissues and other pathological symptoms,the inflammation score was significantly reduced comparing with the model group. Conclusion: The alcohol extract of Elaeagnus angustifolia blossoms could alleviate the respiratory tract inflammation of asthma mice model by reducing contents of IL-4 and IgE,elevating IFN-γ content and regulating immune balance of Th1/Th2.
Objective: To observe the effect of the alcohol extract of Elaeagnus angustifolia blossoms on the respiratory tract inflammation and the serum levels of IL-4,IgE and IFN-γ in the asthmatic mice model,also to explore the mechanism of its inhibition of asthma. Methods: BALB/c mice were randomly divided into 6 groups as followings: the normal group( normal saline),the model group,dexamethasone( 10 mg/kg/d) group and three doses of alcohol extract of Elaeagnus angustifolia blossoms groups( 16 g/kg/d,8 g/kg/d and 4 g/kg/d). Except for the normal group,the asthma models were established in all other groups by improved ovalbumin multipoint injection induced sensitization and atomization inhalation method for 14 d. All mice were sacrificed 24 hours after final stimulation. The pathological changes of lung tissues,counts and classification of white blood cells in bronchoalveolar lavage fluid were observed under microscope,and the inflammatory score was scored. The levels of IL-4,IgE and IFN-γ in bronchoalveolar lavage fluid and serum of mice were determined by double antibody sandwich ELISA. Results: Compared with the model group,the levels of IgE,IL-4 and IL 4/IFN γratio in serum and lavage were significantly decreased in 8 g/kg,16 g/kg/d groups of Elaeagnus angustifolia Blossoms alcohol extract and dexamethasone group in bronchial alveolar asthma model perfusion,the content of IFNγwas significantly increased in 16 g/kg,8 g/kg groups and dexamethasone group in the bronchoalveolar lavage fluid and serum. Compared with the model group,total numbers of white blood cells,eosinophils and mononuclear cell ratio were significantly reduced in Elaeagnus angustifolia blossoms alcohol extract 16 g/kg、8 g/kg group and dexamethasone group,and neutrophil ratio was significantly increased in all alcohol extract groups. Pathological staining showed that the 16 g/kg and 8 g/kg of Elaeagnus angustifolia blossoms alcohol extract significantly reduced the inflammatory infiltration of lung tissues and other pathological symptoms,the inflammation score was significantly reduced comparing with the model group. Conclusion: The alcohol extract of Elaeagnus angustifolia blossoms could alleviate the respiratory tract inflammation of asthma mice model by reducing contents of IL-4 and IgE,elevating IFN-γ content and regulating immune balance of Th1/Th2.
引文
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[2]王妍,李玲,连松刚,等.顶空-固相微萃取气质联用分析宁夏沙枣花中香气成分.中草药,2007,38(11)∶1627～1628.
[3]王妍,王立新.宁夏沙枣花中微量元素的含量分析.食品科技,2004,32(6)∶98～99.
[4]王基云,姚遥,肖旭,等.沙枣花黄酮成分的含量测定及其药理作用的初步研究.时珍国医国药,2010,21(4)∶812～814.
[5]王基云,王丹,姚遥,等.正交实验法优选沙枣花总黄酮提取工艺及沙枣花醇提物体外抗氧化活性的研究.时珍国医国药,2010,21(10)∶2487～2489.
[6]张振华,王基云,姚遥,等.DPPH法测定沙枣花水提物各组分清除自由基的研究.时珍国医国药,2011,22(11)∶2703～2704.
[7]张振华,王基云,姚遥,等.沙枣花超临界二氧化碳萃取物清除自由基的研究.时珍国医国药,2010,21(6)∶1335～1336.
[8]王基云,王丹,姚遥,等.宁夏沙枣花提取物对哮喘模型小鼠氧自由基代谢的影响.宁夏医学杂志,2015,37 (3)∶211～213.
[9]田代印,符州,张燕,等.一种改良型小鼠哮喘模型方法的建立及评价.重庆医科大学学报,2005,30(4)∶601～603.
[10]黄莉,胡玉敏,陈煦艳,等.小鼠哮喘模型的建立及其相关机理初步研究.实验动物与比较医学,2005,25(3)∶140～142.
[11]周玉波,扶招弟,周丽芬,等.硫化氢对臭氧致小鼠气道炎症的影响.中国病理生理杂志,2016,32(10)∶1837～1842.
[12]张羽飞,徐红纳,黄伟,等.川贝母对哮喘模型小鼠气道炎症及ERK/MAPK信号通路的影响.中国药房,2018,29(3)∶343～348.
[13]中华医学会呼吸病学分会哮喘学组.支气管哮喘防治指南(2016年版).中华结核和呼吸杂志.2016,39 (9)∶675～697.
[14]Willskarp M,Nathan A,Page K,et al.New insights into innate immune mechanisms underlying allergenicity.Mucosal Immunology,2010,3(2)∶104～110.
[15]Deo S S,Mistry K J,Kakade A M,et al.Role played by Th2 type cytokines in IgE mediated allergy and asthma.Lung India Official Organ of Indian Chest Society,2010, 27(2)∶66～71.
[16]Robinson D,Hamid Q,Ying S,et al.Prednisolone Treatment in Asthma Is Associated with Modulation of Bronchoalveolar Lavage Cell Interleukin-4, Interleukin-5, and Interferon-γ Cytokine Gene Expression.American Review of Respiratory Disease,1993,148(2)∶401～406.
[17]Grünig,Gabriele,Ford J G,et al.Roles of Interleukin-13 and Interferon-γ in Lung Inflammation.Chest,2002,121(3)∶88S.
[18]Saboonchian F,Jamei R,Sarghein S H.Phenolic and flavonoid content of Elaeagnus angustifolia L. (leaf and flower).Avicenna Journal of Phytomedicine,2014,4(4)∶231～238.
[19]Caliskan E,Elmastas M,Gokce I.Evaluation of antioxidant properties of Elaeagnus angustifolia flowers.Asian Journal of Chemistry,2010,22(4)∶2840～2848.
[20]Bendaikha S,Gadaut, Méredith,Harakat D ,et al.Acylated flavonol glycosides from the flower of Elaeagnus angustifolia L.Phytochemistry,2014,103∶129～136.