Y-27632 is associated with corticosteroid-potentiated control of pulmonary remodeling and inflammation in guinea pigs with chronic allergic inflammation

BMC Pulmonary Medicine

Iolanda de Fátima Lopes Calvo Tibério³  Patrícia Rieken Macedo Rocco¹  Milton de Arruda Martins³  Edna Aparecida Leick³  Carla Máximo Prado³  Mariana Alves Antunes¹  Débora Gonçalves Xisto¹  Anelize Sartori Alves dos Santos³  Adriana Palmeira Dias Rodrigues³  Beatriz Saraiva Romanholo²  Samantha Souza Possa³  Renato Fraga Righetti³  Patricia Angeli Pigati³ 

[1]Laboratory of Pulmonary Investigation, Carlos Chagas Filho Institute of Biophysics, Ilha do Fundão, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil

[2]Institute of Medical Assistance to the State Public Servant of São Paulo (IAMSPE), São Paulo, Brazil

[3]Department of Medicine, School of Medicine, University of São Paulo, São Paulo, Brazil

关键词: Guinea pigs;    Lung mechanics;    Airways;    Y-27632;    Asthma model;    Corticosteroid;    Rho-kinase;   

Others  :  1222562
DOI  :  10.1186/s12890-015-0073-4

 received in 2014-08-27, accepted in 2015-07-06,  发布年份 2015

【 摘 要 】

Background

Previously, we showed that treatment with the Rho-kinase inhibitor Y-27632 was able to control airway responsiveness, inflammation, remodeling, and oxidative stress in an animal model of asthma, suggesting that this drug is beneficial in asthma. However, studies evaluating the effects of these inhibitors in conjunction with corticosteroids on chronic pulmonary inflammation have not been conducted. Therefore, we evaluated the effects of treatment with the Rho-kinase inhibitor Y-27632, with or without concurrent dexamethasone treatment, on airway and lung tissue mechanical responses, inflammation, extracellular matrix remodeling, and oxidative stress in guinea pigs with chronic allergic inflammation.

Methods

The guinea pigs were subjected to seven ovalbumin or saline inhalation exposures. Treatment with Y-27632 (1 mM) and dexamethasone (2 mg/kg) started at the fifth inhalation. Seventy-two hours after the seventh inhalation, the pulmonary mechanics were evaluated and exhaled nitric oxide (E _NO ) levels were determined. The lungs were removed and histological analysis was performed using morphometry.

Results

The treatment of guinea pigs with the Rho-kinase inhibitor and dexamethasone (ORC group) decreased E _NO , the maximal mechanical responses after antigen challenge, inflammation, extracellular matrix remodeling and oxidative stress in the lungs.

This therapeutic strategy reduced the levels of collagen and IFN-γ in the airway walls, as well as IL-2, IFN-γ, 8-iso-PGF2α and NF-κB in the distal parenchyma, when compared to isolated treatment with corticosteroid or Rho-kinase inhibitor (P < 0.05) and reduced the number of TIMP-1-positive cells and eosinophils in the alveolar septa compared to corticosteroid-treated animals (P < 0.05). The combined treatment with the Rho-kinase inhibitor and the corticosteroid provided maximal control over the remodeling response and inflammation in the airways and parenchyma.

Conclusions

Rho-kinase inhibition, alone or in combination with corticosteroids, can be considered a future pharmacological tool for the control of asthma.

【 授权许可】

   
2015 Pigati et al.

【 图 表 】

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【 参考文献 】

• [1]Angeli P, Prado CM, Xisto DG, Silva PL, Pássaro CP, Nakazato HD et al.. Effects of chronic L-NAME treatment lung tissue mechanics, eosinophilic and extracellular matrix responses induced by chronic pulmonary inflammation. Am J Physiol Lung Cell Mol Physiol. 2008; 294:1197-205.
• [2]Bukstein D, Luskin AT, Brooks EA. Exhaled nitric oxide as a tool in managing and monitoring difficult-to-treat asthma. Allergy Asthma Proc. 2011; 32:185-92.
• [3]Chakir J, Shannon J, Molet S, Fukakusa M, Elias J, Laviolette M et al.. Airway remodeling-associated mediators in moderate to severe asthma: effect of steroids on TGF-beta, IL-11, IL-17, and type I and type III collagen expression. J Allergy Clin Immunol. 2003; 111:1293-8.
• [4]Davies DE, Wicks J, Powell RM, Puddicombe SM, Holgate ST. Airway remodeling in asthma: new insights. J Allergy Clin Immunol. 2003; 111:215-25.
• [5]Dolhnikoff M, Morin J, Ludwig MS. Human lung parenchyma responds to contractile stimulation. Am J Respir Crit Care Med. 1998; 158:1607-12.
• [6]Doussau F, Gasman S, Humeau Y, Vitiello F, Popoff M, Boquet P et al.. A Rho-related GTPase is involved in Ca(2+)-dependent neurotransmitter exocytosis. J Biol Chem. 2000; 275:7764-70.
• [7]Fernández AT, Muñoz Cano RM. Importancia clínica de la afectación de la vía aérea pequeña en el asma. Arch Bronconeumol. 2011; 47 Suppl 2:17-19.
• [8]Fredberg JJ, Stamenovic D. On the imperfect elasticity of lung tissue. J Appl Physiol. 1989; 67:2408-19.
• [9]Global Initiative for Asthma (GINA). http://www.ginasthma.org/documents/4. Accessed 20 Jan 2014.
• [10]Goldsmith AM, Hershenson MB, Wolbert MP, Bentley JK. Regulation of airway smooth muscle alpha-actin expression by glucocorticoids. Am J Physiol Lung Cell Mol Physiol. 2007; 292:99-106.
• [11]Goleva E, Hauk PJ, Boguniewicz J, Martin RJ, Leung DY. Airway remodeling and lack of bronchodilator response in steroid-resistant asthma. J Allergy Clin Immunol. 2007; 120:1065-72.
• [12]Gosens R, Schaafsma D, Grootte Bromhaar MM, Vrugt B, Zaagsma J, Meurs H et al.. Growth factor-induced contraction of human bronchial smooth muscle is Rho-kinase-dependent. Eur J Pharmacol. 2004; 494:73-6.
• [13]Hamid Q. Pathogenesis of small airways in asthma. Respiration. 2012; 84:4-11.
• [14]Hashimoto K, Peebles RS, Sheller JR, Jarzecka K, Furlong J, Mitchell DB et al.. Suppression of airway hyperresponsiveness induced by ovalbumin sensitisation and RSV infection with Y-27632, a Rho kinase inhibitor. Thorax. 2002; 57:524-7.
• [15]Hirano K, Derkach DN, Hirano M, Nishimura J, Kanaide H. Protein kinase network in the regulation of phosphorylation and desphosphorylation of smooth muscle myosin light chain. Mol Cell Biochem. 2003; 248:105-14.
• [16]Hoshino M, Takahashi M, Takai Y, Sim J. Inhaled corticosteroids decrease subepithelial collagen deposition by modulation of the balance between matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 expression in asthma. J Allergy Clin Immunol. 1999; 104:356-63.
• [17]Iizuka K, Shimizy Y, Tsukagoshi H, Yoshii A, Harada T, Dobashi K et al.. Evaluation of Y-27632, a rho-kinase inhibitor, as a bronchodilator in guinea pigs. Eur J Pharmacol. 2000; 406:273-79.
• [18]Ito K, Chung KF, Adcock IM. Uptade on glucocorticoid action and resistance. J Allergy Clin Immunol. 2006; 117:522-43.
• [19]Janssen LJ, Premji M, Netherton S, Coruzzi J, Lu-Chao H, Cox PG. Vasoconstrictor actions of isoprostanes via tyrosine kinase and Rho kinase in human and canine pulmonary vascular smooth muscles. Br J Pharmacol. 2001; 132:127-34.
• [20]Jiang J, George SC. TGF-β2 reduces nitric oxide synthase mRNA through a ROCK-dependent pathway in airway epithelial cells. Am J Physiol Lung Cell Mol Physiol. 2011; 301:361-7.
• [21]Kitazawa T, Masuo M, Somlyo AP. G protein-mediated inhibition of myosin light-chain phosphatase in vascular smooth muscle. Proc Natl Acad Sci. 1991; 88:9307-10.
• [22]Kondrikov D, Caldwell RB, Dong Z, Su Y. Reactive oxygen species-dependent RhoA activation mediates collagen synthesis in hyperoxic lung fibrosis. Free Radic Biol Med. 2011; 50:1689-98.
• [23]Kraft M, Part III. Location of asthma inflammation and the distal airways: clinical implications. Curr Med Res Opin. 2007; 23 Suppl 3:21-27.
• [24]Lanças T, Kasahara DI, Prado CM, Tibério IF, Martins MA, Dolhnikoff M. Comparison of early and late responses to antigen of sensitized guinea pig parenchymal lung strips. J Appl Physiol. 2006; 100:1610-6.
• [25]Leick-Maldonado EA, Kay FU, Leonhardt MC, Kasahara DI, Prado CM, Fernandes FT et al.. Comparison of glucocorticoid and cysteinyl leukotriene receptor antagonist treatments in an experimental model of chronic airway inflammation in guinea-pigs. Clin Exp Allergy. 2004; 34:145-152.
• [26]Matsumoto H, Moir LM, Oliver BG, Burgess JK, Roth M, Black JL et al.. Comparison of gel contraction mediated by airway smooth muscle cells from patients with and without asthma. Thorax. 2007; 62:848-54.
• [27]McGown CC, Brown NJ, Hellewell PG, Brookes ZL. ROCK induced inflammation of the microcirculation during endotoxemia mediated by nitric oxide synthase. Microvasc Res. 2011; 81:281-8.
• [28]McMillan SJ, Xanthou G, Lloyd CM. Therapeutic administration of Budesonide ameliorates allergen-induced airway remodelling. Clin Exp Allergy. 2005; 35:388-96.
• [29]Meyer-Schwesinger C, Dehde S, von Ruffer C, Gatzemeier S, Klug P, Wenzel UO et al.. Rho kinase inhibition attenuates LPS-induced renal failure in mice in part by attenuation of NF-kappaB p65 signaling. Am J Physiol Renal Physiol. 2009; 296:1088-99.
• [30]Miller M, Cho JY, McElwain S, Shim JY, Manni M, Baek JS et al.. Corticosteroids prevent myofibroblast accumulation and airway remodeling in mice. Am J Physiol Lung Cell Mol Physiol. 2006; 290:162-9.
• [31]Mong PY, Wang Q. Activation of Rho kinase isoforms in lung endothelial cells during inflammation. J Immunol. 2009; 182:2385-94.
• [32]Mota I, Perini A. A heat labile mercaptoethanol susceptible homocytotropic antibody in the guinea pig. Life Sci II. 1970; 9:923-30.
• [33]Mukai Y, Shimokawa H, Matoba T, Kandabashi T, Satoh S, Hiroki J, et al. Involvement of Rho-kinase in hypertensive vascular disease: a novel therapeutic target in hypertension. FASEB J. 2001;1062–64.
• [34]Nakashima AS, Prado CM, Lanças T, Ruiz VC, Kasahara DI, Leick-Maldonado EA et al.. Oral tolerance attenuates changes in in vitro lung tissue mechanics and extracellular matrix remodeling induced by chronic allergic inflammation in guinea pigs. J Appl Physiol. 2008; 104:1778-85.
• [35]Pfitzer G. Invited review: regulation of myosin phosphorylation in smooth muscle. J Appl Physiol. 2001; 91:497-503.
• [36]Possa SS, Charafeddine HT, Righetti RF, da Silva PA, Almeida-Reis R, Saraiva-Romanholo BM et al.. Rho-kinase inhibition attenuates airway responsiveness, inflammation, matrix remodeling, and oxidative stress activation induced by chronic inflammation. Am J Physiol Lung Cell Mol Physiol. 2012; 303:939-52.
• [37]Prado CM, Leick-Maldonado EA, Kasahara DI, Capelozzi VL, Martins MA, Tibério IF. Effects of acute and chronic nitric oxide inhibition in an experimental model of chronic pulmonary allergic inflammation in guinea pigs. Am J Physiol Lung Cell Mol Physiol. 2005; 289:677-83.
• [38]Prado CM, Leick-Maldonado EA, Yano L, Leme AS, Capelozzi VL, Martins MA et al.. Effects of nitric oxide synthases in chronic allergic airway inflammation and remodeling. Am J Respir Cell Mol Biol. 2006; 35:457-65.
• [39]Prasain N, Stevens T. The actin cytoskeleton in endothelial cell phenotypes. Microvas Res. 2009; 77:53-63.
• [40]Ramos-Barbón D, Parra-Arrondo A. Inflammation and remodeling of the distal airways: studies in humans and experimental models. Arch Bronconeumol. 2011; 47 Suppl 2:2-9.
• [41]Righetti RF, Pigati PA, Possa SS, Habrum FC, Xisto DG, Antunes A et al.. Effects of Rho-kinase inhibition in lung tissue with chronic inflammation. Respir Physiol Neurobiol. 2014; 192:134-46.
• [42]Rocco PR, Negri EM, Kurtz PM, Vasconcellos FP, Silva GH, Capelozzi VL et al.. Lung tissue mechanics and extracellular matrix remodeling in acute lung injury. Am J Respir Crit Care Med. 2001; 164:1067-71.
• [43]Romero PV, Zin WA, Lopez-Aguilar J. Frequency characteristics of lung tissue strip during passive stretch and induced pneumoconstriction. J Appl Physiol. 2001; 91:882-90.
• [44]Ruiz-Schütz VC, Drewiacki T, Nakashima AS, Arantes-Costa FM, Prado CM, Kasahara DI et al.. Oral tolerance attenuates airway inflammation and remodeling in a model of chronic pulmonary allergic inflammation. Respir Physiol Neurobiol. 2009; 165:13-21.
• [45]Sakurada S, Takuwa N, Sugimoto N, Wang Y, Seto M, Sasaki Y et al.. Ca2 + − dependent activation of Rho and Rho kinase in membrane despolarization-induced and receptor stimulation-induced vascular smooth muscle contraction. Cir Res. 2003; 93:548-56.
• [46]Schaafsma D, Bos IS, Zuidhof AB, Zaagsma J, Meurs H. Inhalation of the Rho-kinase inhibitor Y-27632 reverses allergen-induced airway hyperresponsiveness after the early and late asthmatic reaction. Respir Res. 2006; 7:121. BioMed Central Full Text
• [47]Schaafsma D, Gosens R, Bos IS, Meurs H, Zaagsma J, Nelemans SA. Allergic sensitization enhances the contribution of Rho-kinase to airway smooth muscle contraction. Br J Pharmacol. 2004; 143:477-84.
• [48]Souza FC, Gobbato NB, Maciel RG, Prado CM, Martins MA, Leick EA et al.. Effects of corticosteroid, montelukast and iNOS inhibition on distal lung with chronic inflammation. Respir Physiol Neurobiol. 2013; 185:435-45.
• [49]Starling CM, Prado CM, Leick-Maldonado EA, Lanças T, Reis FG, Aristóteles LR et al.. Inducible nitric oxide synthase inhibition attenuates lung tissue responsiveness and remodeling in a model of chronic pulmonary inflammation in guinea pigs. Respir Physiol Neurobiol. 2009; 165:185-94.
• [50]Taki F, Kume H, Kobayashi T, Ohta H, Aratake H, Shimokata K. Effects of Rho-kinase inactivation on eosinophilia and hyper-reactivity in murine airways by allergen challenges. Clin Exp Allergy. 2007; 37:599-607.
• [51]Tibério IF, Turco GM, Leick-Maldonado EA, Sakae RS, Paiva SO, do Patrocinio M et al.. Effects of neurokinin depletion on airway inflammation induced by chronic antigen exposure. Am J Respir Crit Care Med. 1997; 155:1739-47.
• [52]Uehata M, Ishizaki T, Satoh H, Ono T, Kawahara T, Morishita T et al.. Calcium sensitization of smooth muscle mediated by a Rho-associated protein kinase in hypertension. Nature. 1997; 389:990-4.
• [53]Weibel ER. The challenge of measuring lung structure. On the "Standards for the Quantitative Assessment of Lung Structure”. Nihon Kokyuki Gakkai Zasshi. 2010; 48:637-43.
• [54]Zhou H, Zhang KX, Li YJ, Guo BY, Wang M, Wang M. Fasudil hydrochloride hydrate, a Rho-kinase inhibitor, suppresses high glucose-induce proliferation and collagen synthesis in rat cardiac fibroblasts. Clin Exp Pharmacol Physiol. 2011; 38:387-94.