姜黄素衍生物WZ01对哮喘气道炎症介质IL-13和组胺及糖皮质激素受体的影响
|
摘要
目的:研究姜黄素衍生物WZ01对哮喘气道炎症反应、白细胞介素(IL)-13、组胺及糖皮质激素受体(GR)的影响,探讨WZ01抑制哮喘气道炎症的机制。方法:30只小鼠,随机分为:正常对照组、哮喘组、低剂量WZ01组、中剂量WZ01组、高剂量WZ01组,每组6只。在第21~第27天1%卵清白蛋白(OVA)雾化激发,每次雾化前30 min予腹腔注射相应剂量0.9%氯化钠溶液或WZ01。末次激发24 h内,麻醉并处死小鼠,留取肺组织标本HE染色检测炎症病理改变,留取支气管肺泡灌洗液(BALF)进行细胞分类计数,ELISA法检测IL-13和组胺浓度,Western blot和免疫组织化学染色检测肺组织GR蛋白的表达。结果:与正常对照组比较,哮喘组小鼠气道周围可见明显炎症细胞浸润及黏液分泌,BALF中炎症细胞数量、IL-13及组胺水平均显著升高(P<0.01)。与哮喘组相比,各剂量WZ01组气道周围炎症细胞浸润及黏液分泌显著减少,BALF中淋巴细胞和嗜酸性粒细胞数量明显减少,BALF中IL-13和组胺浓度显著下降,差异均有统计学意义(P<0.01);BALF中淋巴细胞数量与IL-13和组胺浓度均呈显著正相关(r值分别为0.886、0.886,P均<0.01),嗜酸性粒细胞数量与IL-13和组胺浓度均呈显著正相关(r值分别为0.897、0.898,P均<0.01)。Western blot和免疫组织化学染色结果显示,哮喘小鼠肺组织GR表达明显降低,与哮喘组相比,不同剂量WZ01均能显著上调哮喘小鼠肺组织GR表达水平。结论:姜黄素衍生物WZ01可显著抑制哮喘气道炎症细胞浸润和黏液分泌,可能与其降低炎症介质IL-13和组胺浓度及上调哮喘小鼠肺组织GR表达水平有关,WZ01可能有望成为防治哮喘气道炎症的新药。
Objective: To investigate the effects of curcumin derivative WZ01 on asthmatic airway inflammation, inflammatory mediators IL-13 and histamine, and glucocorticoid receptor(GR). Methods: Thirty mice were randomly divided into: control group, asthma group, low dose WZ01 group, moderate dose WZ01 group and high dose WZ01 group. Each group was 6 mice, respectively. After sensitized with OVA, the mice were challenging with 1% OVA from the 21 th to 27 th day. Each mouse was intraperitoneally injected with corresponding drugs or normal saline half an hour before atomizing. After the last challenging, mice were anesthetized and sacrificed within 24 hours, lung tissues were obtained for detecting inflammatory alteration with hematoxylin-eosin staining, and the BALF for cellular classification and counting and the concentrations of IL-13 and histamine by ELISA assay, the expression of GR in lung tissues was detected with immumohistochemical staining and Western blot. Results: Compared with the normal control group, HE staining showed massive inflammatory cellular infiltration and mucous secretion accumulated in the asthma group, ELISA analysis showed significantly increased number of inflammatory cells and concentrations of IL-13 and histamine in the BALF(P<0.01). Compared with the asthma group, each dose of WZ01 markedly inhibited the inflammatory cellular infiltration and mucous secretion in the airway; In each dose WZ01 group, the number of lymphocytes and eosinophils in BALF were distinctly decreased(P<0.01), accompanied by the remarkable decline of the concentrations of IL-13 and histamine(P<0.01); the number of lymphocytes had remarkably positive relationships with the concentrations of IL-13 and histamine in the BALF(r values were 0.886 and 0.886, respectively, P<0.01). The number of esoinophils had also remarkably positive relationships with the concentrations of IL-13 and histamine in the BALF(r values were 0.897 and 0.898, respectively, P<0.01). The results of immumohistochemical staining and Western blot showed that GR expression in the lung tissues of asthmatic mice was decrease. Compare with the asthma group, different doses of WZ01 could predominantly reverse and up-regulate the expression of GR in the lung tissues of asthmatic mice. Conclusion: WZ01 can significantly inhibit the inflammatory cellular infiltration and mucus secretion in asthma, which may be attributable to the decline of the inflammatory mediators IL-13 and histamine and the up-regulated expression of GR in the lung tissues of asthmatic mice. WZ01 may be a new drug of prevention and treatment for airway inflammation in asthma.
Objective: To investigate the effects of curcumin derivative WZ01 on asthmatic airway inflammation, inflammatory mediators IL-13 and histamine, and glucocorticoid receptor(GR). Methods: Thirty mice were randomly divided into: control group, asthma group, low dose WZ01 group, moderate dose WZ01 group and high dose WZ01 group. Each group was 6 mice, respectively. After sensitized with OVA, the mice were challenging with 1% OVA from the 21 th to 27 th day. Each mouse was intraperitoneally injected with corresponding drugs or normal saline half an hour before atomizing. After the last challenging, mice were anesthetized and sacrificed within 24 hours, lung tissues were obtained for detecting inflammatory alteration with hematoxylin-eosin staining, and the BALF for cellular classification and counting and the concentrations of IL-13 and histamine by ELISA assay, the expression of GR in lung tissues was detected with immumohistochemical staining and Western blot. Results: Compared with the normal control group, HE staining showed massive inflammatory cellular infiltration and mucous secretion accumulated in the asthma group, ELISA analysis showed significantly increased number of inflammatory cells and concentrations of IL-13 and histamine in the BALF(P<0.01). Compared with the asthma group, each dose of WZ01 markedly inhibited the inflammatory cellular infiltration and mucous secretion in the airway; In each dose WZ01 group, the number of lymphocytes and eosinophils in BALF were distinctly decreased(P<0.01), accompanied by the remarkable decline of the concentrations of IL-13 and histamine(P<0.01); the number of lymphocytes had remarkably positive relationships with the concentrations of IL-13 and histamine in the BALF(r values were 0.886 and 0.886, respectively, P<0.01). The number of esoinophils had also remarkably positive relationships with the concentrations of IL-13 and histamine in the BALF(r values were 0.897 and 0.898, respectively, P<0.01). The results of immumohistochemical staining and Western blot showed that GR expression in the lung tissues of asthmatic mice was decrease. Compare with the asthma group, different doses of WZ01 could predominantly reverse and up-regulate the expression of GR in the lung tissues of asthmatic mice. Conclusion: WZ01 can significantly inhibit the inflammatory cellular infiltration and mucus secretion in asthma, which may be attributable to the decline of the inflammatory mediators IL-13 and histamine and the up-regulated expression of GR in the lung tissues of asthmatic mice. WZ01 may be a new drug of prevention and treatment for airway inflammation in asthma.
引文
[1]全国儿科哮喘协作组.第三次中国城市儿童哮喘流行病学调查[J].中华儿科杂志,2013,51(10):729-735.
[2]胡晓光,张海邻,李昌崇,等.温州城区哮喘儿童流行病学调查[J].温州医科大学学报,2014,44(11):804-810.
[3]BORRIELLO F,LONGO M,SPINELLI R,et al.IL-3 synergises with basophil-derived IL-4 and IL-13 to promote the alternative activation of human monocytes[J].Eur J Immunol,2015,45(7):2042-2051.
[4]RANDALL M J,KOSTIN S F,BURGESS E J,et al.Antiinflammatory effects of levalbuterol-induced 11beta-hydroxysteroid dehydrogenase type 1 activity in airway epithelial cells[J].Front Endocrinol(Lausanne),2015,5:236.
[5]HU G X,LIN H,LIAN Q Q,et al.Curcumin as a potent and selective inhibitor of 11beta-hydroxysteroid dehydrogenase1:improving lipid profiles in high-fat-diet-treated rats[J].PLo S One,2013,8(3):e49976.
[6]CHONG L,ZHANG W,NIE Y,et al.Protective effect of curcumin on acute airway inflammation of allergic asthma in mice through Notch1-GATA3 signaling pathway[J].Inflammation,2014,37(5):1476-1485.
[7]MOON D O,KIM M O,LEE H J,et al.Curcumin attenuates ovalbumin-induced airway inflammation by regulating nitric oxide[J].Biochem Biophys Res Commun,2008,375(2):275-279.
[8]OH S W,CHA J Y,JUNG J E,et al.Curcumin attenuates allergic airway inflammation and hyper-responsiveness in mice through NF-kappa B inhibition[J].J Ethnopharmacol,2011,136(3):414-421.
[9]YASUKAWA A,HOSOKI K,TODA M,et al.Eosinophils promote epithelial to mesenchymal transition of bronchial epithelial cells[J].PLo S One,2013,8(5):e64281.
[10]GON Y,ITO R,HATTORI T,et al.Serum eosinophil-derived neurotoxin:Correlation with persistent airflow limitation in adults with house-dust mite allergic asthma[J].Allergy Asthma Proc,2015,36(6):113-120.
[11]DRAKE L Y,IIJIMA K,HARA K,et al.B cells play key roles in th2-type airway immune responses in mice exposed to natural airborne allergens[J].PLo S One,2015,10(3):e0121660.
[12]BARTEMES K R,KEPHART G M,FOX S J,et al.Enhanced innate type 2 immune response in peripheral blood from patients with asthma[J].J Allergy Clin Immunol,2014,134(3):671-678.
[13]HU A,FATMA S,CAO J,et al.Th2 cytokine-induced upregulation of 11beta-hydroxysteroid dehydrogenase-1 facilitates glucocorticoid suppression of proasthmatic airway smooth muscle function[J].Am J Physiol Lung Cell Mol Physiol,2009,296(5):L790-803.
[14]LEE M Y,SEO C S,LEE N H,et al.Anti-asthmatic effect of schizandrin on OVA-induced airway inflammation in a murine asthma model[J].Int Immunopharmacol,2010,10(11):1374-1379.
[15]OESER K,MAXEINER J,SYMOWSKI C,et al.T cells are the critical source of IL-4/IL-13 in a mouse model of allergic asthma[J].Allergy,2015,70(11):1440-1449.
[16]ASHRAF M I,SHAHZAD M,SHABBIR A.Oxyresveratrol ameliorates allergic airway inflammation via attenuation of IL-4,IL-5,and IL-13 expression levels[J].Cytokine,2015,76(2):375-381.
[17]OH C K,GEBA G P,MOLFINO N.Investigational therapeutics targeting the IL-4/IL-13/STAT-6 pathway for the treatment of asthma[J].Eur Respir Rev,2010,19(115):46-54.
[18]MAY R D,FUNG M.Strategies targeting the IL-4/IL-13 axes in disease[J].Cytokine,2015,75(1):89-116.
[19]TSURIKISAWA N,OSHIKATA C,TSUBURAI T,et al.Physiologic airway responses to inhale histamine and acetylcholine in patients with mild asthma as analyzed by forced osciliation[J].Arerugi,2015,64(7):952-970.
[20]BENKO R,MOLNAR T F,SZOMBATI V,et al.Combined inhibition of histamine H1 receptors and leukotrienes reduces compound 48/80-induced contraction of the human bronchus in vitro[J].Pharmacology,2015,96(5-6):253-255.
[21]HORIE M,SAITO A,YAMAUCHI Y,et al.Histamine induces human lung fibroblast-mediated collagen gel contraction via histamine H1 receptor[J].Exp Lung Res,2014,40(5):222-236.
[22]SILVA R A,ALMEIDA F M,OLIVO C R,et al.Exercise reverses OVA-induced inhibition of glucocorticoid receptor and increases anti-inflammatory cytokines in asthma[J].Scand J Med Sci Sports,2016,26(1):82-92.
[23]CORNEJO S,TANTISIRA K,RABY B A,et al.Nuclear bioavailability of the glucocorticoid receptor in a pediatric asthma cohort with variable corticosteroid responsiveness[J].Pediatr Res,2015,78(5):505-512.
[24]JOSHI T,JOHNSON M,NEWTON R,et al.The long-acting beta2-adrenoceptor agonist,indacaterol,enhances glucocorticoid receptor-mediated transcription in human airway epithelial cells in a gene-and agonist-dependent manner[J].Br J Pharmacol,2015,172(10):2634-2653.
[2]胡晓光,张海邻,李昌崇,等.温州城区哮喘儿童流行病学调查[J].温州医科大学学报,2014,44(11):804-810.
[3]BORRIELLO F,LONGO M,SPINELLI R,et al.IL-3 synergises with basophil-derived IL-4 and IL-13 to promote the alternative activation of human monocytes[J].Eur J Immunol,2015,45(7):2042-2051.
[4]RANDALL M J,KOSTIN S F,BURGESS E J,et al.Antiinflammatory effects of levalbuterol-induced 11beta-hydroxysteroid dehydrogenase type 1 activity in airway epithelial cells[J].Front Endocrinol(Lausanne),2015,5:236.
[5]HU G X,LIN H,LIAN Q Q,et al.Curcumin as a potent and selective inhibitor of 11beta-hydroxysteroid dehydrogenase1:improving lipid profiles in high-fat-diet-treated rats[J].PLo S One,2013,8(3):e49976.
[6]CHONG L,ZHANG W,NIE Y,et al.Protective effect of curcumin on acute airway inflammation of allergic asthma in mice through Notch1-GATA3 signaling pathway[J].Inflammation,2014,37(5):1476-1485.
[7]MOON D O,KIM M O,LEE H J,et al.Curcumin attenuates ovalbumin-induced airway inflammation by regulating nitric oxide[J].Biochem Biophys Res Commun,2008,375(2):275-279.
[8]OH S W,CHA J Y,JUNG J E,et al.Curcumin attenuates allergic airway inflammation and hyper-responsiveness in mice through NF-kappa B inhibition[J].J Ethnopharmacol,2011,136(3):414-421.
[9]YASUKAWA A,HOSOKI K,TODA M,et al.Eosinophils promote epithelial to mesenchymal transition of bronchial epithelial cells[J].PLo S One,2013,8(5):e64281.
[10]GON Y,ITO R,HATTORI T,et al.Serum eosinophil-derived neurotoxin:Correlation with persistent airflow limitation in adults with house-dust mite allergic asthma[J].Allergy Asthma Proc,2015,36(6):113-120.
[11]DRAKE L Y,IIJIMA K,HARA K,et al.B cells play key roles in th2-type airway immune responses in mice exposed to natural airborne allergens[J].PLo S One,2015,10(3):e0121660.
[12]BARTEMES K R,KEPHART G M,FOX S J,et al.Enhanced innate type 2 immune response in peripheral blood from patients with asthma[J].J Allergy Clin Immunol,2014,134(3):671-678.
[13]HU A,FATMA S,CAO J,et al.Th2 cytokine-induced upregulation of 11beta-hydroxysteroid dehydrogenase-1 facilitates glucocorticoid suppression of proasthmatic airway smooth muscle function[J].Am J Physiol Lung Cell Mol Physiol,2009,296(5):L790-803.
[14]LEE M Y,SEO C S,LEE N H,et al.Anti-asthmatic effect of schizandrin on OVA-induced airway inflammation in a murine asthma model[J].Int Immunopharmacol,2010,10(11):1374-1379.
[15]OESER K,MAXEINER J,SYMOWSKI C,et al.T cells are the critical source of IL-4/IL-13 in a mouse model of allergic asthma[J].Allergy,2015,70(11):1440-1449.
[16]ASHRAF M I,SHAHZAD M,SHABBIR A.Oxyresveratrol ameliorates allergic airway inflammation via attenuation of IL-4,IL-5,and IL-13 expression levels[J].Cytokine,2015,76(2):375-381.
[17]OH C K,GEBA G P,MOLFINO N.Investigational therapeutics targeting the IL-4/IL-13/STAT-6 pathway for the treatment of asthma[J].Eur Respir Rev,2010,19(115):46-54.
[18]MAY R D,FUNG M.Strategies targeting the IL-4/IL-13 axes in disease[J].Cytokine,2015,75(1):89-116.
[19]TSURIKISAWA N,OSHIKATA C,TSUBURAI T,et al.Physiologic airway responses to inhale histamine and acetylcholine in patients with mild asthma as analyzed by forced osciliation[J].Arerugi,2015,64(7):952-970.
[20]BENKO R,MOLNAR T F,SZOMBATI V,et al.Combined inhibition of histamine H1 receptors and leukotrienes reduces compound 48/80-induced contraction of the human bronchus in vitro[J].Pharmacology,2015,96(5-6):253-255.
[21]HORIE M,SAITO A,YAMAUCHI Y,et al.Histamine induces human lung fibroblast-mediated collagen gel contraction via histamine H1 receptor[J].Exp Lung Res,2014,40(5):222-236.
[22]SILVA R A,ALMEIDA F M,OLIVO C R,et al.Exercise reverses OVA-induced inhibition of glucocorticoid receptor and increases anti-inflammatory cytokines in asthma[J].Scand J Med Sci Sports,2016,26(1):82-92.
[23]CORNEJO S,TANTISIRA K,RABY B A,et al.Nuclear bioavailability of the glucocorticoid receptor in a pediatric asthma cohort with variable corticosteroid responsiveness[J].Pediatr Res,2015,78(5):505-512.
[24]JOSHI T,JOHNSON M,NEWTON R,et al.The long-acting beta2-adrenoceptor agonist,indacaterol,enhances glucocorticoid receptor-mediated transcription in human airway epithelial cells in a gene-and agonist-dependent manner[J].Br J Pharmacol,2015,172(10):2634-2653.