Jo Eu-Ri, Youn Cha Kyung, Jun Yonghyun, Cho Sung Il
Department of Otolaryngology-Head and Neck Surgery, Chosun University College of Medicine, Gwangju, South Korea.
Department of Premedical Science, Chosun University College of Medicine, Gwangju, South Korea.
Int J Pediatr Otorhinolaryngol. 2019 May;120:30-35. doi: 10.1016/j.ijporl.2019.02.001. Epub 2019 Feb 4.
While cisplatin is an effective chemotherapeutic agent, it can cause irreversible hearing loss. Ototoxicity leads to dose reduction during the cisplatin chemotherapy and results in inadequate treatment of malignant tumors. This study aimed to investigate the protective effects of ferulic acid on cisplatin-induced ototoxicity.
House Ear Institute-Organ of Corti 1 (HEI-OC1) cells were exposed to 30 μM of cisplatin for 24 h with or without pretreatment with ferulic acid. Cell viability was determined using the WST assay. Apoptotic cells were identified using TUNEL assay. Western blot analysis was performed to examine the change in expression of cleaved caspase, cleaved poly-ADP-ribose polymerase (PARP), nuclear factor erythroid 2-related factor 2 (Nrf2), and catalase. Intracellular reactive oxygen species (ROS) were determined by flow cytometry. Real-time PCR analyses were performed to examine the mRNA levels of antioxidant enzymes including glutamate-cysteine ligase catalytic subunit (Gclc), glutathione peroxidase 2 (Gpx2), catalase, and superoxide dismutase 2 (SOD2). Phalloidin staining of the organ of Corti was performed to determine hair cell survival or degeneration.
Pretreatment with ferulic acid before cisplatin exposure significantly increased cell viability, levels of antioxidant enzymes, and hair cell survival. In addition, pretreatment with ferulic acid significantly reduced apoptotic cells, levels of cleaved caspase, levels of cleaved PARP, and intracellular ROS production.
Our results demonstrated that ferulic acid inhibited cisplatin-induced cytotoxicity by preventing ROS formation and inducing the production of endogenous antioxidants and indicated that ferulic acid might be used as a protective agent against cisplatin-induced ototoxicity.
虽然顺铂是一种有效的化疗药物，但它可导致不可逆的听力损失。耳毒性会导致顺铂化疗期间剂量减少，从而导致恶性肿瘤治疗不充分。本研究旨在探讨阿魏酸对顺铂所致耳毒性的保护作用。
将耳科研究所-柯蒂氏器1（HEI-OC1）细胞暴露于30μM顺铂中24小时，顺铂暴露前有无阿魏酸预处理。使用WST法测定细胞活力。使用TUNEL法鉴定凋亡细胞。进行蛋白质免疫印迹分析以检测裂解的半胱天冬酶、裂解的聚ADP核糖聚合酶（PARP）、核因子红细胞2相关因子2（Nrf2）和过氧化氢酶表达的变化。通过流式细胞术测定细胞内活性氧（ROS）。进行实时PCR分析以检测抗氧化酶的mRNA水平，包括谷氨酸-半胱氨酸连接酶催化亚基（Gclc）、谷胱甘肽过氧化物酶2（Gpx2）、过氧化氢酶和超氧化物歧化酶2（SOD2）。对柯蒂氏器进行鬼笔环肽染色以确定毛细胞的存活或退化情况。
顺铂暴露前用阿魏酸预处理可显著提高细胞活力、抗氧化酶水平和毛细胞存活率。此外，阿魏酸预处理可显著减少凋亡细胞、裂解的半胱天冬酶水平、裂解的PARP水平和细胞内ROS生成。
我们的结果表明，阿魏酸通过阻止ROS形成和诱导内源性抗氧化剂的产生来抑制顺铂诱导的细胞毒性，并表明阿魏酸可能用作预防顺铂诱导的耳毒性的保护剂。
