克拉霉素对激素反应性通路中组蛋白去乙酰化酶2及糖皮质激素受体的影响
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摘要
目的观察克拉霉素对激素反应性通路中组蛋白去乙酰化酶2(HDAC2)及糖皮质激素受体(GR)的影响,探讨克拉霉素对烟雾暴露哮喘小鼠可能的治疗作用。方法选择BALB/c小鼠作为研究对象,通过卵蛋白致敏激发建立哮喘模型组(OVA组)、烟雾暴露哮喘组(SEA组)以及哮喘接触烟雾暴露后通过克拉霉素治疗组(CAM组),同时设立对照组平行比较。采用组织病理学检测各组小鼠肺组织炎症情况,并通过PCR检测肺组织中HDAC2mRNA的表达情况和Western blot检测肺组织中HDAC2和GR蛋白的表达。结果组织病理学观察OVA组和SEA组均存在肺组织气道炎症,其中SEA组纤毛被严重破坏,而CAM组炎症细胞浸润减弱。与CAM组相比,SEA组小鼠支气管肺泡灌洗液中IL-4和IL-8水平均显著提高(104.36±14.39 vs.65.49±10.82、681.35±66.18 vs.321.49±90.37,P=0.031、0.017)。CAM组HDAC2mRNA表达显著高于SEA组(0.062±0.013 vs.0.031±0.015,P=0.032)。Western blot检测结果显示,与CAM组相比,SEA组HDAC2蛋白(0.23±0.017 vs.0.49±0.022,P=0.033)和GR蛋白(0.19±0.014 vs.0.64±0.023,P=0.011)表达均显著降低。结论克拉霉素能抑制烟雾暴露的哮喘小鼠气道炎症反应,作用机制可能通过与GR相结合,影响下游HDAC2基因表达有关。
Objective To explore the effects of clarithromycin on the expressions of histone deacetylase-2(HDAC2)and glucocorticoid receptor(GR)of cigarette smoke-exposed asthmatic mice.Methods BALB/c mice were chosen to be the subjects of this study.They were raised to establish asthma model(OVA group);and mice in one asthma group were exposed to smoke(SEA group),one asthma group were treated with clarithromycin(CAM group)after smoke exposure.Control group mice were used as parallel comparison.The histopathological changes were studied to assess lung tissue inflammation.Cell counts in bronchoalveolar lavage fluid were also tested for airway inflammation.Histone deacelytase2(HDAC2)activity of lung tissues was measured by qRT-PCR.HDAC2 and GR expressions in the lung tissue were detected by Western blot.Results Histopathologic observation showed massive infiltration of inflammatory cells in both OVA group and SEA group,while inflammation infiltration attenuated in CAM group.Compared with those in CAM group,the levels of IL-4 and IL-8 in bronchoalveolar lavage fluid of SEA group increased significantly(104.36±14.39 vs.65.49±10.82,681.35±66.18 vs.321.49±90.37;P=0.031,0.017).The expression of HDAC2 mRNA in CAM group was significantly higher than that in SEA group(0.062±0.013 vs.0.031±0.015,P=0.032).The expressions of HDAC2 protein(0.23±0.017 vs.0.49±0.022,P=0.033)and GR protein(0.19±0.014 vs.0.64±0.023,P=0.011)were significantly lower in SEA group than in CAM group.Conclusion Clarithromycin could attenuate airway inflammation in smokeexposed asthmatic mice.The mechanism of action may be related to the expression of HDAC2 gene in the lower reaches by combining with GR.
Objective To explore the effects of clarithromycin on the expressions of histone deacetylase-2(HDAC2)and glucocorticoid receptor(GR)of cigarette smoke-exposed asthmatic mice.Methods BALB/c mice were chosen to be the subjects of this study.They were raised to establish asthma model(OVA group);and mice in one asthma group were exposed to smoke(SEA group),one asthma group were treated with clarithromycin(CAM group)after smoke exposure.Control group mice were used as parallel comparison.The histopathological changes were studied to assess lung tissue inflammation.Cell counts in bronchoalveolar lavage fluid were also tested for airway inflammation.Histone deacelytase2(HDAC2)activity of lung tissues was measured by qRT-PCR.HDAC2 and GR expressions in the lung tissue were detected by Western blot.Results Histopathologic observation showed massive infiltration of inflammatory cells in both OVA group and SEA group,while inflammation infiltration attenuated in CAM group.Compared with those in CAM group,the levels of IL-4 and IL-8 in bronchoalveolar lavage fluid of SEA group increased significantly(104.36±14.39 vs.65.49±10.82,681.35±66.18 vs.321.49±90.37;P=0.031,0.017).The expression of HDAC2 mRNA in CAM group was significantly higher than that in SEA group(0.062±0.013 vs.0.031±0.015,P=0.032).The expressions of HDAC2 protein(0.23±0.017 vs.0.49±0.022,P=0.033)and GR protein(0.19±0.014 vs.0.64±0.023,P=0.011)were significantly lower in SEA group than in CAM group.Conclusion Clarithromycin could attenuate airway inflammation in smokeexposed asthmatic mice.The mechanism of action may be related to the expression of HDAC2 gene in the lower reaches by combining with GR.
引文
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[22]周霞,党亚洁,王桂芳,等.烟曲霉暴露对支气管哮喘大鼠肺组织糖皮质激素受体表达的调控作用[J].中华结核和呼吸杂志,2017,40(3):182-187.
[23]KADIYALA V,SASSE SK,ALTONSY MO,et al.Cistrome analysis of glucocorticoid receptor activity in bronchial epithelial cells defines novel mechanisms of steroid efficacy[J].Ann Am Thorac Soc,2016,13(Suppl 1):S103.
[2]MULEY P,SHAH M,MULEY A.Omega-3 fatty acids supplementation in children to prevent asthma:Is it worthy?—A systematic review and meta-analysis[J].J Allergy,2015,2015(2):312052.
[3]SHAKURNIA AH,ASSAR S,AFRA M,et al.Prevalence of asthma among schoolchildren in Ahvaz,Islamic Republic of Iran[J].East Mediterr Health J,2010,16(6):651-656.
[4]FITZGERALD JM,FOUCART S,COOYLE S,et al.Montelukast as add-on therapy to inhaled corticosteroids in the management of asthma(the SAS trial)[J].Can Respir J,2016,16(Suppl A):5A-14A.
[5]BARNES CB,ULRIKLRIK CS.Asthma and adherence to inhaled corticosteroids:Current status and future perspectives[J].Respir Care,2015,60(3):455-468.
[6]PRUTEANU AI,CHAUHAN BF,ZHANG L,et al.Inhaled corticosteroids in children with persistent asthma:Is there a dose response impact on growth?—an overview of Cochrane reviews[J].Paediatr Respir Rev,2015,16(1):51-52.
[7]CHEN X,KANG YB,WANG LQ,et al.Addition to inhaled corticosteroids of leukotriene receptor antagonists versus theophylline for symptomatic asthma:A meta-analysis[J].J Thorac Dis,2015,7(4):644-652.
[8]SPEARS M,DONNELLY I,JOLLY L,et al.Effect of lowdose theophylline plus beclometasone on lung function in smokers with asthma:apilot study[J].Eur Respir J,2009,33(5):1010-1017.
[9]王文,吴小军.大环内酯类抗生素在慢性气道炎症疾病治疗中的进展[J].临床肺科杂志,2016,21(7):1338-1341.
[10]TONG X,GUO T,LIU S,et al.Macrolide antibiotics for treatment of asthma in adults:A meta-analysis of 18 randomized controlled clinical studies[J].Pulm Pharmacol Ther,2015,31(5):99-108.
[11]LUISI F,GANDOLFI TD,DAUDT AD,et al.Anti-inflammatory effects of macrolides in childhood lung diseases[J].J Bras Pneumol,2012,38(6):786-796.
[12]REITER J,DEMIREL N,MENDY A,et al.Macrolides for the long-term management of asthma—A meta-analysis of randomized clinical trials[J].Allergy,2013,68(8):1040–1049.
[13]陈金龙,庄思斯,秦玉明,等.大环内酯类药物治疗肺炎支原体感染患儿变异性哮喘的疗效观察[J].中华医院感染学杂志,2015,25(23):5374-5376.
[14]JR GJ,ROLLINS DR,MARTIN RJ.Macrolides in the treatment of asthma[J].Curr Opin Pulm Med,2012,18(1):76-84.
[15]COEMAN M,VAN DY,BAUTERS F,et al.Neomacrolides in the treatment of patients with severe asthma and/or bronchiectasis:A retrospective observational study[J].Ther Adv Respir Dis,2011,5(6):377-386.
[16]YAO X,WANG W,LI Y,et al.Characteristics of IL-25 and allergen-induced airway fibrosis in a murine model of asthma[J].Respirology,2015,20(5):730-738.
[17]NAGASAKI T,MATSUMOTO H.Influences of smoking and aging on allergic airway inflammation in asthma[J].Allergol Int,2013,62(2):171-179.
[18]SHIMODA T,OBASE Y,KISHIKAWA R,et al.Influence of cigarette smoking on airway inflammation and inhaled corticosteroid treatment in patients with asthma[J].Allergy Asthma Proceedings,2016,37(4):50-58.
[19]KESHAVARZ A R S,MONIRI R,SAFFARI M,et al.The Helicobacter pylori resistance rate to clarithromycin in Iran[J].Microb Drug Resist,2015,21(1):69-73.
[20]CI X,XIAO C,XU X,et al.Short-term roxithromycin treatment attenuates airway inflammation via MAPK/NF-κB activation in a mouse model of allergic asthma[J].Inflamm Res,2012,61(7):749-758.
[21]BASYIGIT I,YILDIZ F,OZKARA SK,et al.The effect of clarithromycin on inflammatory markers in chronic obstructive pulmonary disease:preliminary data[J].Ann Pharmacother,2004,38(9):1400-1405.
[22]周霞,党亚洁,王桂芳,等.烟曲霉暴露对支气管哮喘大鼠肺组织糖皮质激素受体表达的调控作用[J].中华结核和呼吸杂志,2017,40(3):182-187.
[23]KADIYALA V,SASSE SK,ALTONSY MO,et al.Cistrome analysis of glucocorticoid receptor activity in bronchial epithelial cells defines novel mechanisms of steroid efficacy[J].Ann Am Thorac Soc,2016,13(Suppl 1):S103.