기니픽 제1형 전정유모세포에서 젠타마이신에 의해 유발되는 전정독성에 대한 Epigallocatechin-3-Gallate의 보호 효과
The Protective Effect of Epigallocatechin-3-Gallate Against Gentamicin Vestibular Ototoxicity in Type I Vestibular Hair Cell of Guinea Pig
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Background and Objectives Gentamicin (GM) is well known for its vestibulotoxicity. There have been many reports about vestibulotoxicity, however, its mechanism is still unclear. So far, it is known that GM affects the voltage-dependent K+ current and nitric oxide (NO) production. Epigallocatechin-3-gallate (EGCG) is the major component of green tea and is knownto have anti-oxidative and anti-toxic effect. This study was undertaken to investigate the protective effect of EGCG against gentamicin on vestibular hair cell (VHC). Materials and Method White guinea pigs (200-250 g) were rapidly decapitated and thetemporal bones were immediately removed. Under a dissecting microscope, the crista ampullaris was obtained. The dissociated VHCs were transferred into a recording chamber mountedonto an inverted microscope. Whole-cell membrane currents and potentials were recorded using standard patch-clamp techniques. In addition, measurements of NO production were obtained using the NO-sensitive dye, 4,5-diamino-fluorescein diacetate (DAF-2DA). Results Type I VHCs Voltage-dependent K+ current was activated from low depolarizingstimulation. As the stimulation increased, higher current was detected. Voltage-dependent K+current in type I VHCs was decreased when GM (200 μM) was administrated and GM effectsof K+ current inhibition was significantly blocked by EGCG. Extracellular GM induced an increase in DAF-2DA fluorescence, which thus indicates NO production in VHCs. Also, the GMinduced NO production was inhibited by EGCG. Conclusion GM inhibits voltage-dependent K+ current by releasing NO in isolated type IVHCs. EGCG blocks this inhibitory effects, suggesting a protective role on GM vestibulotoxicity. Korean J Otorhinolaryngol-Head Neck Surg 2014;57(4):226-32
Abstract
Background and Objectives Gentamicin (GM) is well known for its vestibulotoxicity. There have been many reports about vestibulotoxicity, however, its mechanism is still unclear. So far, it is known that GM affects the voltage-dependent K+ current and nitric oxide (NO) production. Epigallocatechin-3-gallate (EGCG) is the major component of green tea and is knownto have anti-oxidative and anti-toxic effect. This study was undertaken to investigate the protective effect of EGCG against gentamicin on vestibular hair cell (VHC). Materials and Method White guinea pigs (200-250 g) were rapidly decapitated and thetemporal bones were immediately removed. Under a dissecting microscope, the crista ampullaris was obtained. The dissociated VHCs were transferred into a recording chamber mountedonto an inverted microscope. Whole-cell membrane currents and potentials were recorded using standard patch-clamp techniques. In addition, measurements of NO production were obtained using the NO-sensitive dye, 4,5-diamino-fluorescein diacetate (DAF-2DA). Results Type I VHCs Voltage-dependent K+ current was activated from low depolarizingstimulation. As the stimulation increased, higher current was detected. Voltage-dependent K+current in type I VHCs was decreased when GM (200 μM) was administrated and GM effectsof K+ current inhibition was significantly blocked by EGCG. Extracellular GM induced an increase in DAF-2DA fluorescence, which thus indicates NO production in VHCs. Also, the GMinduced NO production was inhibited by EGCG. Conclusion GM inhibits voltage-dependent K+ current by releasing NO in isolated type IVHCs. EGCG blocks this inhibitory effects, suggesting a protective role on GM vestibulotoxicity. Korean J Otorhinolaryngol-Head Neck Surg 2014;57(4):226-32
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