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KMID : 0361020070500050385
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Korean Journal of Otolaryngology - Head and Neck Surgery
2007 Volume.50 No. 5 p.385 ~ p.390
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The Effect of IP3 Receptor Inhibition Using 2-APB on Gentamicin Ototoxicity in Cochlear Sensory Cell
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Moon Jeong-Hwan
Jung Jae-Yun
Lim Eun-Suk
Rhee Chung-Ku
Ahn Jin-Chul
Seong Yeon-Sun
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Abstract
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Background & Objectives: Elevated intracellular calcium level is known to play important roles in the apoptotic pathway. IP3 receptor (ligand-gated channels that release Ca2+ from intracellular stores) is emerging as a key site for regulation of apoptosis. 2-Aminoethoxydiphenyl borate (2-APB) is one of the reliable IP3 receptor antagonists. We examined the effect of 2-APB on gentamicin ototoxicity in vitro, using the HEI-OC1 cell line.
Materials & Methods: HEI-OC1 cells were trWWeated with 100micrometer gentamicin. Using a CaspACE assay, we measured the caspases-3 activity in the gentamicin treated hair cells with and without 2-APB pre-incubation. We also observed intra-cellular calcium concentrations in HEI-OC1 cells using a confocal microscopy (calcium green-1 stain). Live cell imaging was performed by using fluorescence video-time lapse system.
Results: Cytosolic calcium elevation by gentamicin was remarkably inhibited by 2-APB. Caspases-3 activities of gentamicin treated cells were higher than those of the control. After incubation with 2-APB, caspases-3 activities and cell death of gentamicin treated cells were shown to decrease.
Conclusion: 2-APB reduces Caspases-3 activity in the gentamicin treated HEI-OC1 cells by inhibition of cytosolic calcium increase.
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KEYWORD
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2-APB, Caspase-3, Gentamicin, Calcium, IP3 Receptor
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