• 南通大學(xué)附屬醫(yī)院呼吸科( 江蘇南通 226001) ;;
  • 2 南京醫(yī)科大學(xué)第一附屬醫(yī)院呼吸科( 江蘇南京 210029)通訊作者: 殷凱生, E-mail: yinks@ 126 . com;

目的  構(gòu)建β2 受體( β2R)下調(diào)哮喘動(dòng)物模型, 以研究β2R下調(diào)及其激素保護(hù)作用的機(jī)制。方法  32 只BALB/ c 小鼠隨機(jī)分成對(duì)照組、哮喘組、β2R下調(diào)組和地塞米松組, 每組8 只。哮喘組、β2R下調(diào)組和地塞米松組分別于實(shí)驗(yàn)的第0、14和21 d 腹腔注射卵白蛋白( OVA) 和氫氧化鋁的混懸液200 μL, 第28 d 開(kāi)始連續(xù)1 周( 30 min/d) 霧化吸入1% OVA( W/V) 溶液;β2R下調(diào)組和地塞米松組在最后1 周每天霧化吸入OVA 之前腹腔注射60 μg 沙丁胺醇及霧化吸入0. 01% 沙丁胺醇30 min; 地塞米松組同時(shí)予以腹腔注射地塞米松5 mg·kg-1·d - 1 , 連續(xù)7 d。對(duì)照組腹腔注射等量的磷酸鹽緩沖液( PBS) , 以PBS霧化吸入。用體積描記箱測(cè)定各組小鼠氣道阻力。測(cè)定BALF 中白細(xì)胞總數(shù)和分類(lèi)計(jì)數(shù)。ELISA 法檢測(cè)BALF 中IL-4、IFN-γ以及血清總IgE 濃度。提取小鼠肺組織總蛋白, 免疫印跡實(shí)驗(yàn)測(cè)定β2R總量, 放射配基結(jié)合實(shí)驗(yàn)測(cè)定β2膜受體數(shù)量。結(jié)果  哮喘組、β2R下調(diào)
組與對(duì)照組、地塞米松組比較, 高劑量乙酰膽堿激發(fā)下的氣道阻力明顯增高( P  lt;0. 01) , BALF 中嗜酸粒細(xì)胞、中性粒細(xì)胞和淋巴細(xì)胞明顯增多( P lt;0. 01) , BALF中IL-4 和血清總IgE 水平顯著增高( P lt;0. 01) , BALF 中IFN-γ水平顯著降低( P lt;0. 01) 。β2R下調(diào)組肺組織β2R 總量和β2 膜受體數(shù)量均顯著低于其余三組( P lt;0. 01) , 對(duì)照組、哮喘組、地塞米松組之間無(wú)顯著差異。結(jié)論  在傳統(tǒng)的OVA 致敏激發(fā)哮喘動(dòng)物模型基礎(chǔ)上, 采用腹腔注射結(jié)合霧化吸入沙丁胺醇的方法成功構(gòu)建了β2R下調(diào)的哮喘動(dòng)物模型。地塞米松對(duì)β2R下調(diào)具有保護(hù)作用。

引用本文： 劉華,倪松石,蔡和群,張樺,殷凱生. β2 受體下調(diào)哮喘模型的構(gòu)建研究. 中國(guó)呼吸與危重監(jiān)護(hù)雜志, 2010, 9(2): 122-127. doi: 復(fù)制

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2. . Mak JC,Nishikawa M,Shirasaki H,et al.Protective effects of a glucocorticoid on downregulation of pulmonary beta 2-adrenergic receptors in vivo.J Clin Invest,1995,96:99-106.
3. .Liu H,Zhou LF,Zhang Q,et al.Increased RhoGDI2 and peroxiredoxin 5 levels in asthmatic murine model of beta2-adrenoceptor desensitization: a proteomics approach.Chin Med J (Engl), 2008,121:355-362.
4. .Vroon A,Lombardi MS,Kavelaars A,et al.Taxol normalizes the impaired agonist-induced beta2-adrenoceptor internalization in splenocytes from GRK2+/- mice.Eur J Pharmacol,2007,560:9-16.
5. .Moore RH,Millman EE,Godines V,et al.Salmeterol stimulation dissociates beta2-adrenergic receptor phosphorylation and internalization.Am J Respir Cell Mol Biol,2007,36:254-261.
6. .Finney PA,Belvisi MG,Donnelly LE,et al.Albuterol-induced downregulation of Gsalpha accounts for pulmonary beta(2)-adrenoceptor desensitization in vivo.J Clin Invest,2000,106:125-135.
7. .Zhao LL,Lötvall J,Lindén A,et al.Prolonged eosinophil production after allergen exposure in IFN-gammaR KO mice is IL-5 dependent.Scand J Immunol,2008,67:480-488.
8. .Taher YA,van Esch BC,Hofman GA,et al.1alpha,25-dihydroxyvitamin D3 potentiates the beneficial effects of allergen immunotherapy in a mouse model of allergic asthma:role for IL-10 and TGF-beta.J Immunol,2008,180:5211-5221.
9. .Mousavi T, Salek Moghadam A, Falak R. Immunotherapy of Chenopodium album induced asthma by intranasal administration of CpG oligodeoxynucleotides in BALB/c mice.Iran J Immunol,2008,5:57-63.
10. .Rovati GE,Baroffio M,Citro S,et al.Cysteinyl-leukotrienes in the regulation of beta2-adrenoceptor function: an in vitro model of asthma.Respir Res,2006,7:103.
11. .Penn RB,Shaver JR,Zangrilli JG,et al.Effects of inflammation and acute beta-agonist inhalation on beta 2-AR signaling in human airways.Am J Physiol,1996,271:L601-608.
12. .Chen CL,Wang YM,Liu CF,et al.The effect of water-soluble chitosan on macrophage activation and the attenuation of mite allergen-induced airway inflammation.Biomaterials,2008,29:2173-2182.
13. .Mathur SK,Schwantes EA,Jarjour NN,et al.Age-related changes in eosinophil function in human subjects.Chest,2008,133:412-419.
14. .Wheway J,Herzog H,Mackay F.NPY and receptors in immune and inflammatory diseases.Curr Top Med Chem,2007,7:1743-1752.
15. .Moon KA,Kim SY,Kim TB,et al.Allergen-induced CD11b+ CD11c(int) CCR3+ macrophages in the lung promote eosinophilic airway inflammation in a mouse asthma model.Int Immunol,2007,19:1371-1381.
16. .MacKinnon AC,Farnworth SL,Hodkinson PS,et al.Regulation of alternative macrophage activation by galectin-3.J Immunol,2008,180:2650-2658.
17. .Hermanowicz-Salamon J,Domaga?a-Kulawik J,Maskey-Warzechowska M,et al.Macrophage phenotype in induced sputum in asthma subjects.Pneumonol Alergol Pol,2006,74:101-105.
18. .Fitzpatrick AM,Holguin F,Teague WG,et al.Alveolar macrophage phagocytosis is impaired in children with poorly controlled asthma.J Allergy Clin Immunol,2008,121:1372-1378.
19. .Esnault S,Rosenthal LA,Shen ZJ,et al.A critical role for Pin1 in allergic pulmonary eosinophilia in rats.J Allergy Clin Immunol,2007,120:1082-1088.
20. .Lee TH,Arm JP.The effects of a fish oil enriched diet on experimentally-induced and clinical asthma.Adv Prostaglandin Thromboxane Leukot Res,1989,19:606-609.
21. .Paggiaro PL,Dente FL,Azzarà A,et al.Abnormal migratory activity of peripheral neutrophils from asthmatic patients and its modulation by inhaled glucocorticoids.J Investig Allergol Clin Immunol,1993,3:237-244.
22. .Zhao J,Zhu H,Wong CH,,et al.Increased lungkine and chitinase levels in allergic airway inflammation: a proteomics approach.Proteomics,2005,5:2799-2807.
23. .Macdowell AL,Peters SP.Neutrophils in asthma.Curr Allergy Asthma Rep,2007,7:464-468.
24. .Elekes K,Helyes Z,Kereskai L,et al.Inhibitory effects of synthetic somatostatin receptor subtype 4 agonists on acute and chronic airway inflammation and hyperreactivity in the mouse.Eur J Pharmacol,2008,578:313-322.
25. .Melgert BN,Timens W,Kerstjens HA,et al.Effects of 4 months of smoking in mice with ovalbumin-induced airway inflammation.Clin Exp Allergy,2007,37:1798-1808.
26. .Zacharasiewicz A,Wilson N,Lex C,et al.Clinical use of noninvasive measurements of airway inflammation in steroid reduction in children.Am J Respir Crit Care Med,2005,171:1077-1082.
27. .Nejman-Gryz P,Grubek-Jaworska H,Glapiński J,et al.Effects of the phosphodiestrase-4 inhibitor rolipram on lung resistance and inflammatory reaction in experimental asthma.J Physiol Pharmacol,2006,57:229-239.
  1. 1. .Shahzeidi S,Aujla PK,Nickola TJ,et al.Temporal analysis of goblet cells and mucin gene expression in murine models of allergic asthma.Exp lung Res,2003,29: 549-565.
  2. 2. . Mak JC,Nishikawa M,Shirasaki H,et al.Protective effects of a glucocorticoid on downregulation of pulmonary beta 2-adrenergic receptors in vivo.J Clin Invest,1995,96:99-106.
  3. 3. .Liu H,Zhou LF,Zhang Q,et al.Increased RhoGDI2 and peroxiredoxin 5 levels in asthmatic murine model of beta2-adrenoceptor desensitization: a proteomics approach.Chin Med J (Engl), 2008,121:355-362.
  4. 4. .Vroon A,Lombardi MS,Kavelaars A,et al.Taxol normalizes the impaired agonist-induced beta2-adrenoceptor internalization in splenocytes from GRK2+/- mice.Eur J Pharmacol,2007,560:9-16.
  5. 5. .Moore RH,Millman EE,Godines V,et al.Salmeterol stimulation dissociates beta2-adrenergic receptor phosphorylation and internalization.Am J Respir Cell Mol Biol,2007,36:254-261.
  6. 6. .Finney PA,Belvisi MG,Donnelly LE,et al.Albuterol-induced downregulation of Gsalpha accounts for pulmonary beta(2)-adrenoceptor desensitization in vivo.J Clin Invest,2000,106:125-135.
  7. 7. .Zhao LL,Lötvall J,Lindén A,et al.Prolonged eosinophil production after allergen exposure in IFN-gammaR KO mice is IL-5 dependent.Scand J Immunol,2008,67:480-488.
  8. 8. .Taher YA,van Esch BC,Hofman GA,et al.1alpha,25-dihydroxyvitamin D3 potentiates the beneficial effects of allergen immunotherapy in a mouse model of allergic asthma:role for IL-10 and TGF-beta.J Immunol,2008,180:5211-5221.
  9. 9. .Mousavi T, Salek Moghadam A, Falak R. Immunotherapy of Chenopodium album induced asthma by intranasal administration of CpG oligodeoxynucleotides in BALB/c mice.Iran J Immunol,2008,5:57-63.
  10. 10. .Rovati GE,Baroffio M,Citro S,et al.Cysteinyl-leukotrienes in the regulation of beta2-adrenoceptor function: an in vitro model of asthma.Respir Res,2006,7:103.
  11. 11. .Penn RB,Shaver JR,Zangrilli JG,et al.Effects of inflammation and acute beta-agonist inhalation on beta 2-AR signaling in human airways.Am J Physiol,1996,271:L601-608.
  12. 12. .Chen CL,Wang YM,Liu CF,et al.The effect of water-soluble chitosan on macrophage activation and the attenuation of mite allergen-induced airway inflammation.Biomaterials,2008,29:2173-2182.
  13. 13. .Mathur SK,Schwantes EA,Jarjour NN,et al.Age-related changes in eosinophil function in human subjects.Chest,2008,133:412-419.
  14. 14. .Wheway J,Herzog H,Mackay F.NPY and receptors in immune and inflammatory diseases.Curr Top Med Chem,2007,7:1743-1752.
  15. 15. .Moon KA,Kim SY,Kim TB,et al.Allergen-induced CD11b+ CD11c(int) CCR3+ macrophages in the lung promote eosinophilic airway inflammation in a mouse asthma model.Int Immunol,2007,19:1371-1381.
  16. 16. .MacKinnon AC,Farnworth SL,Hodkinson PS,et al.Regulation of alternative macrophage activation by galectin-3.J Immunol,2008,180:2650-2658.
  17. 17. .Hermanowicz-Salamon J,Domaga?a-Kulawik J,Maskey-Warzechowska M,et al.Macrophage phenotype in induced sputum in asthma subjects.Pneumonol Alergol Pol,2006,74:101-105.
  18. 18. .Fitzpatrick AM,Holguin F,Teague WG,et al.Alveolar macrophage phagocytosis is impaired in children with poorly controlled asthma.J Allergy Clin Immunol,2008,121:1372-1378.
  19. 19. .Esnault S,Rosenthal LA,Shen ZJ,et al.A critical role for Pin1 in allergic pulmonary eosinophilia in rats.J Allergy Clin Immunol,2007,120:1082-1088.
  20. 20. .Lee TH,Arm JP.The effects of a fish oil enriched diet on experimentally-induced and clinical asthma.Adv Prostaglandin Thromboxane Leukot Res,1989,19:606-609.
  21. 21. .Paggiaro PL,Dente FL,Azzarà A,et al.Abnormal migratory activity of peripheral neutrophils from asthmatic patients and its modulation by inhaled glucocorticoids.J Investig Allergol Clin Immunol,1993,3:237-244.
  22. 22. .Zhao J,Zhu H,Wong CH,,et al.Increased lungkine and chitinase levels in allergic airway inflammation: a proteomics approach.Proteomics,2005,5:2799-2807.
  23. 23. .Macdowell AL,Peters SP.Neutrophils in asthma.Curr Allergy Asthma Rep,2007,7:464-468.
  24. 24. .Elekes K,Helyes Z,Kereskai L,et al.Inhibitory effects of synthetic somatostatin receptor subtype 4 agonists on acute and chronic airway inflammation and hyperreactivity in the mouse.Eur J Pharmacol,2008,578:313-322.
  25. 25. .Melgert BN,Timens W,Kerstjens HA,et al.Effects of 4 months of smoking in mice with ovalbumin-induced airway inflammation.Clin Exp Allergy,2007,37:1798-1808.
  26. 26. .Zacharasiewicz A,Wilson N,Lex C,et al.Clinical use of noninvasive measurements of airway inflammation in steroid reduction in children.Am J Respir Crit Care Med,2005,171:1077-1082.
  27. 27. .Nejman-Gryz P,Grubek-Jaworska H,Glapiński J,et al.Effects of the phosphodiestrase-4 inhibitor rolipram on lung resistance and inflammatory reaction in experimental asthma.J Physiol Pharmacol,2006,57:229-239.