RESUMEN
The new allele B*54:39 showed one nucleotide difference from B*54:01:01 in codon 10 (ACC->AGC).
Asunto(s)
Pueblo Asiatico/genética , Antígenos HLA-B/genética , Prueba de Histocompatibilidad/métodos , Polimorfismo de Nucleótido Simple , Análisis de Secuencia de ADN/métodos , Alelos , Sustitución de Aminoácidos , Secuencia de Bases , Humanos , Homología de SecuenciaRESUMEN
OBJECTIVE: Caffeic acid phenethyl ester (CAPE) is known to reduce the generation of oxygen-derived free radicals, which is a major mechanism of aminoglycoside-induced ototoxicity. The objective of the present study was to evaluate the effects of CAPE on neomycin-induced ototoxicity in zebrafish (Brn3c: EGFP). METHODS: Five-day post-fertilization zebrafish larvae (n=10) were exposed to 125 µM neomycin and one of the following CAPE concentrations for 1h: 50, 100, 250, 500, or 1000 µM. Ultrastructural changes were evaluated using scanning electron microscopy (SEM). The terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick-end labeling (TUNEL) assay and 2-[4-(dimethylamino)styryl]-N-ethylpyridiniumiodide (DASPEI) assay were performed for evaluation of apoptosis and mitochondrial damage. RESULTS: CAPE decreased neomycin-induced hair cell loss in the neuromasts (500 µM CAPE: 12.7 ± 1.1 cells, 125 µM neomycin only: 6.3 ± 1.1 cells; n = 10, P < 0.05). In the ultrastructural analysis, structures of mitochondria and hair cells were preserved when exposed to 125 µM neomycin and 500 µM CAPE. CAPE decreased apoptosis and mitochondrial damage. CONCLUSION: In the present study, CAPE attenuated neomycin-induced hair cell damage in zebrafish. The results of the current study suggest that neomycin induces apoptosis, and the apoptotic cell death can be prevented by treatment with CAPE in zebrafish.
Asunto(s)
Antibacterianos/toxicidad , Antioxidantes/farmacología , Ácidos Cafeicos/farmacología , Células Ciliadas Auditivas/efectos de los fármacos , Neomicina/toxicidad , Alcohol Feniletílico/análogos & derivados , Animales , Apoptosis/efectos de los fármacos , Células Ciliadas Auditivas/patología , Microscopía Electrónica de Rastreo , Mitocondrias/efectos de los fármacos , Mitocondrias/patología , Modelos Animales , Alcohol Feniletílico/farmacología , Pez CebraRESUMEN
NecroX-5, one of the derivatives of NecroX series compounds, is a mitochondrial reactive oxygen species and reactive nitrogen species scavenger that inhibits cell death against various kinds of oxidative stresses. The objective of the present study was to evaluate the effects of NecroX-5 on neomycin-induced ototoxicity in transgenic zebrafish (Brn3C: EGFP). Five days post-fertilization, zebrafish larvae were exposed to 125 µM neomycin and one of the following NecroX-5 concentrations for 1 h: 10, 25, 50, and 75 µM. Hair cells within the neuromasts of the supraorbital (SO1 and SO2), otic (O1), and occipital (OC1) lateral lines were analyzed using fluorescence microscopy (n = 10). The terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay and 2-[4-(dimethylamino) styryl]-N-ethylpyridiniumiodide (DASPEI) assay were performed for evaluation of apoptosis and mitochondrial damage. Ultrastructural changes were evaluated using scanning electron microscopy. NecroX-5 decreased neomycin-induced hair cell loss in the neuromasts (NecroX-5 50 µM: 13.4 ± 2.0 cells, 125 µM neomycin only: 8.1 ± 1.2 cells; n = 10, P < 0.05) and decreased the TUNEL reaction. The ultrastructural analysis showed that the structures of mitochondria and hair cells within the neuromasts were preserved in zebrafish exposed to 125 µM neomycin and 50 µM NecroX-5. NecroX-5 decreased apoptosis and mitochondrial damage. In conclusion, NecroX-5 attenuated neomycin-induced hair cell loss in zebrafish.
