Sung Kyun Kim1, Gi Jung Im1, Yun Suk An2, Se Hee Lee1, Hak Hyun Jung1, Sang Yoo Park3. 1. Department of Otolaryngology-Head and Neck Surgery, Korea University College of Medicine, Inchon-ro 73, Seongbuk-Gu, Seoul, Republic of Korea. 2. Department of Otolaryngology-Head and Neck Surgery, Jesaeng Hospital, Seohyeon-ro 20, Bundang-Gu, Seognam, Gyeonggi, Republic of Korea. 3. Department of Otolaryngology-Head and Neck Surgery, Wonju College of Medicine, Yonsei University, 162 Ilsan-dong, Wonju 220-701, Republic of Korea. Electronic address: sangyoo3@yonsei.ac.kr.
Abstract
CONCLUSION: D-α-tocopherol succinate significantly reduced a cisplatin-induced hair cell loss in HEI-OC1 cell lines. These effects were mediated by its scavenging activity against reactive oxygen species (ROS) and inhibition of apoptosis. OBJECTIVES: Alpha-tocopherol is a class of methylated phenols, known as fat-soluble antioxidants, and is a different form of vitamin E, which reduces free radicals and acts as an antioxidant. We hypothesized that the antioxidative effect of α-tocopherol could protect against cisplastin-induced cytotoxicity, and thus evaluated its effects on cisplatin-induced ototoxicity in HEI-OC1 auditory cells. METHODS: HEI-OC1 cells were pretreated with D-α-tocopherol succinate at a concentration of 10 µM for 24 h, and then exposed to 15 µM cisplatin for 48 h. The cellular viability was measured by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. The intracellular ROS level was measured by using a fluorescent dye, 2',7'-dichlorofluorescein diacetate (DCFH-DA). Both Annexin V-FITC and propidium iodide (PI) staining were performed to analyze the pattern of apoptosis. The enzymatic activity of caspase-3 was assayed with caspase3/CPP32 fluorometric assay kit. Also, it was assessed by immunoblotting technique of poly-ADP-ribose polymerase (PARP). RESULTS: Pretreatment with 10 µM D-α-tocopherol succinate protected HEI-OC1 auditory cells against cisplatin-induced cytotoxicity. D-α-tocopherol succinate significantly reduced the cisplatin-induced increase in ROS. D-α-tocopherol succinate treatment induced a 15% reduction of ROS and 50% decrease in necrosis and late apoptosis as compared to cisplatin treatment. D-α-tocopherol succinate also decreased the activation of caspase-3 and reduced levels of cleaved poly-ADP-ribose polymerase (PARP).
CONCLUSION: D-α-tocopherol succinate significantly reduced a cisplatin-induced hair cell loss in HEI-OC1 cell lines. These effects were mediated by its scavenging activity against reactive oxygen species (ROS) and inhibition of apoptosis. OBJECTIVES:Alpha-tocopherol is a class of methylated phenols, known as fat-soluble antioxidants, and is a different form of vitamin E, which reduces free radicals and acts as an antioxidant. We hypothesized that the antioxidative effect of α-tocopherol could protect against cisplastin-induced cytotoxicity, and thus evaluated its effects on cisplatin-induced ototoxicity in HEI-OC1 auditory cells. METHODS: HEI-OC1 cells were pretreated with D-α-tocopherol succinate at a concentration of 10 µM for 24 h, and then exposed to 15 µM cisplatin for 48 h. The cellular viability was measured by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. The intracellular ROS level was measured by using a fluorescent dye, 2',7'-dichlorofluorescein diacetate (DCFH-DA). Both Annexin V-FITC and propidium iodide (PI) staining were performed to analyze the pattern of apoptosis. The enzymatic activity of caspase-3 was assayed with caspase3/CPP32 fluorometric assay kit. Also, it was assessed by immunoblotting technique of poly-ADP-ribose polymerase (PARP). RESULTS: Pretreatment with 10 µM D-α-tocopherol succinate protected HEI-OC1 auditory cells against cisplatin-induced cytotoxicity. D-α-tocopherol succinate significantly reduced the cisplatin-induced increase in ROS. D-α-tocopherol succinate treatment induced a 15% reduction of ROS and 50% decrease in necrosis and late apoptosis as compared to cisplatin treatment. D-α-tocopherol succinate also decreased the activation of caspase-3 and reduced levels of cleaved poly-ADP-ribose polymerase (PARP).
Authors: Anna I Petridou; Eleftheria T Zagora; Petros Petridis; George S Korres; Maria Gazouli; Ioannis Xenelis; Efthymios Kyrodimos; Georgia Kontothanasi; Andriana C Kaliora Journal: Nutrients Date: 2019-12-12 Impact factor: 5.717