ZELİHA KAPUSUZ GENCER, Levent SAYDAM, A. Noam COHEN, Cemal CİNGİ

N-acetylcysteine effects on sinonasal cilia function

Amaç: N-asetil sisteinin (NAC) insan respiratuvar epitelyal hücre kültüründe, başta elektrolit transportu ve siliyer atım frekansı olmak üzere farmakolojik etkilerinin değerlendirilmesi. Yöntem: Hava-sıvı arayüzünde üretilmiş, iyi diferansiye edilmiş insan bronşiyal epitelyal hücre kültüründe farklı konsantrasyonlarda NAC uygulaması yapıldı. İdeal siliyer atım frekansını sağlayan NAC konsantrasyonu saptandı. N-asetil sisteinin atım hızına etkisi saptandıktan sonra, etkinliği ATP veya IBMX ile araştırılarak etki mekanizması ortaya konmaya çalışıldı. Siliyer atım frekansı Sissons-Ammons video analiz sistemi ile saptandı. Bulgular: Siliyer fonksiyonlarda maksimum stimülasyonun 10 mg/ml NAC konsantrasyonunda olduğu saptandı. Yıkama sonrası siliyer hareketler dramatik olarak arttı. Bu artış NAC artı IBMX ve NAC artı ATP yıkamalarından sonra daha da artış gösterdi. Sonuç: N-asetil sisteinin apikal kullanımı, siliyer atım frekansını belirgin düzeyde artırmaktadır. N-asetil sistein sonrası PBS ile yıkamak ile klinik etkinlik elde edilebilir.

N-asetil sisteinin sinonazal siliyer atım frekansına etkileri

Objective: To evaluate the pharmacological effects of the N-acetylcysteine (NAC) on human respiratory epithelial cultures specifically addressing electrolyte transport and cilia beat frequency. Methods: Well-differentiated human bronchial epithelial cultures grown at an air liquid interface were treated on the apical or basolateral surface with varying concentrations of NAC. The best NAC concentration for ideal cilia beat frequency was found. The effects of NAC were evaluated on cilia beat frequency. After the effect of N-acetylcysteine on beat rate was found, its efficiency was investigated by ATP or IBMX to understand its mechanism of action. Changes in ciliary beat frequency were determined using the Sissons-Ammons video analysis system. Results: Maximal stimulatory effect on cilia function was evident at 10 mg/ml NAC concentration. After wash up, cilia movement were increased very dramatically. This increase of cilia beat frequency was even higher after NAC plus IBMX and NAC plus ATP washings. Conclusion: Apical application of NAC prominently stimulates cilia beat frequency and after wash up, cilia movement was increased very dramatically. After NAC use by washing with PBS in clinical efficacy can be enhanced.

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1. Gibbons IR. The relationship between the fine structure and direction of beat in gill cilia of a lamellibranch mollusc. J Biophys Biochem Cytol 1961;11:179205.
2. Chmiel JF, Davis PB. State of the art: why do the lungs of patients with cystic fibrosis become infected and why cant they clear the infection? Respir Res 2003;4:8.
3. Boucher RC. Regulation of airway surface liquid volume by human airway epithelia. Pflugers Arch 2003;445:4958.
4. Tarran R, Button B, Boucher RC. Regulation of normal and cys- tic fibrosis airway surface liquid volume by phasic shear stress. Annu Rev Physiol 2006;68:54361.
5. King M. Mucus, mucociliary clearance and coughing. In: Bates DV, editor. Respiratory function in disease. 3rd ed. Philadelphia, PA: Saunders; 1989. p. 6978.
6. Boogaard R, de Jongste J C, Merkus PJ . Pharmacotherapy of impaired mucociliary clearance in non-CF pediatric lung disease. A review of the literature. Pediatr Pulmonol 2007;42:9891001.
7. Gillissen A, Nowak D. Characterization of N-acetylcysteine and ambroxol in anti-oxidant therapy. Respir Med 1998;92:60923.
8. Köttgen M, Busch AE, Hug MJ , Greger R, Kunzelmann K. N- Acetyl-L-cysteine and its derivatives activate a Cl- conductance in epithelial cells. Pflugers Arch 1996;431:54955.
9. Rochat T, Lacroix J S, Jornot L. N-acetylcysteine inhibits Na+ absorption across human nasal epithelial cells. J Cell Physiol 2004; 201:10616.
10. Andersson C, Gaston B, Roomans GM. S-Nitrosoglutathione induces functional DeltaF508-CFTR in airway epithelial cells. Biochem Biophys Res Commun 2002;297:5527.
11. Zaman K, Palmer LA, Doctor A, Hunt JF, Gaston B. Concentration-dependent effects of endogenous S-nitrosoglu- tathione on gene regulation by specificity proteins Sp3 and Sp1. Biochem J 2004;380(Pt 1):6774.
12. Servetnyk Z, Krjukova J , Gaston B, et al. Activation of chloride transport in CF airway epithelial cell lines and primary CF nasal epithelial cells by S-nitrosoglutathione. Respir Res 2006;7:124.
13. Ventresca GP, Cicchetti V, Ferrari V. Acetylcysteine. In: Braga PC, Allegra L, editors. Drugs in bronchial mucology. New York, NY: Raven Press; 1989. p. 77102.
14. Stafanger G, Bisgaard H, Pedersen M, Mørkassel E, Koch C. Effect of N-acetylcysteine on the human nasal ciliary activity in vitro. Eur J Respir Dis 1987;70:15762.
15. Garcia GJ , Picher M, Zuo P, et al. Computational model for the regulation of extracellular ATP and adenosine in airway epithelia. Subcell Biochem 2011;55:5174.
16. Milara J , Armengot M, Mata M, Morcillo EJ , Cortijo J . Role of adenylate kinase type 7 expression on cilia motility: possible link in primary ciliary dyskinesia. Am J Rhinol Allergy 2010;24:1815.
17. Woodworth BA, Zhang S, Tamashiro E, Bhargave G, Palmer JN, Cohen NA. Zinc increases ciliary beat frequency in a calcium- dependent manner. Am J Rhinol Allergy 2010;24:610.
18. Varelogianni1 G. Oliynyk I, Roomans G. M, Johannesson M. The effect of N-acetylcysteine on chloride efflux from airway epithelial cells. Cell Biol Int 2010;34:24552.