ENT Institute and Department of Otorhinolaryngology, Eye & ENT Hospital, State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, NHC Key Laboratory of Hearing Medicine, Fudan University, Shanghai, China.
Department of Otorhinolaryngology-Head and Neck Surgery, First Affiliated Hospital of Nanchang University, Nanchang, China.
CNS Neurosci Ther. 2024 Feb;30(2):e14403. doi: 10.1111/cns.14403. Epub 2023 Aug 14.
Cinchonine (CN) and its isomer cinchonidine (CD), two of the common cinchona alkaloids, are wildly used as antimalarial drugs. However, the effects of CN and CD on the auditory system are unknown.
Molecular docking and molecular dynamics (MD) simulation were used for predicting effective drugs. The CCK-8 assay was conducted for assessing cell viability in House Ear Institute-Organ of Corti 1 (HEI-OC1) cells. MitoSox Red staining revealed reactive oxygen species (ROS) amounts. TMRM staining was used to assess the mitochondrial membrane potential (ΔΨm). Immunofluorescence staining of myosin 7a was used to examine hair cells (HCs) in cisplatin-treated neonatal mouse cochlear explants, while TUJ-1 immunostaining was used for the detection of spiral ganglion neurons (SGNs). Cleaved caspase-3 and TUNEL immunostaining were utilized for apoptosis assessment. Immunoblot was carried out to detect PI3K-AKT signaling effectors.
Pretreatment with CN or CD significantly increased cell viability and reduced mitochondrial dysfunction and ROS accumulation in cisplatin-treated HEI-OC1 cells. Immunofluorescent staining of cochlear explants showed that CN and CD attenuated cisplatin-induced damage to SGNs and HCs. Immunoblot revealed that CN and CD downregulated the expression of cleaved caspase-3 and activated PI3K-AKT signaling in cisplatin-injured HEI-OC1 cells.
CD and CN can reduce ototoxicity caused by cisplatin and might help treat cisplatin-associated hearing loss.
金鸡纳生物碱中的辛可宁（CN）和其异构体辛可尼丁（CD）是两种常用的抗疟药物。然而，CN 和 CD 对听觉系统的影响尚不清楚。
采用分子对接和分子动力学（MD）模拟预测有效药物。CCK-8 法检测 House Ear Institute-Organ of Corti 1（HEI-OC1）细胞活力。MitoSox Red 染色法检测活性氧（ROS）含量。TMRM 染色法评估线粒体膜电位（ΔΨm）。肌球蛋白 7a 的免疫荧光染色用于检测顺铂处理的新生小鼠耳蜗外植体中的毛细胞（HCs），而 TUJ-1 免疫染色用于检测螺旋神经节神经元（SGNs）。用 cleaved caspase-3 和 TUNEL 免疫染色检测细胞凋亡。免疫印迹法检测 PI3K-AKT 信号通路效应物。
CN 或 CD 预处理可显著提高顺铂处理的 HEI-OC1 细胞的活力，减少线粒体功能障碍和 ROS 积累。耳蜗外植体的免疫荧光染色显示，CN 和 CD 可减轻顺铂诱导的 SGNs 和 HCs 损伤。免疫印迹显示，CN 和 CD 下调了顺铂损伤的 HEI-OC1 细胞中 cleaved caspase-3 的表达并激活了 PI3K-AKT 信号通路。
CD 和 CN 可减轻顺铂引起的耳毒性，并可能有助于治疗顺铂相关性听力损失。
