NAD+对顺铂所致毛细胞氧化应激损伤的拮抗作用及相关基因表达调控

doi:10.6040/j.issn.1673-3770.0.2025.054

摘要/Abstract

摘要： 目的探究烟酰胺腺嘌呤二核苷酸(nicotinamide adenine dinucleotide, NAD⁺)对顺铂(cisplatin, Cis)诱导耳毒性的拮抗效果和机制。方法体外实验采用HEI-OC1细胞系和出生后3 d的C57BL/6J乳鼠耳蜗基底膜培养模型,评估NAD⁺对Cis毒性的拮抗作用;在体实验通过鼓室注射Cis和NAD⁺于C57BL/6J成年小鼠,并进行听觉脑干反应测试;采用转录组测序分析HEI-OC1细胞中Cis组与Cis⁺NAD⁺组之间的差异基因;定量聚合酶链式反应对关键基因进行验证。结果 NAD⁺能够显著提高HEI-OC1细胞在Cis处理下的细胞活力,并有效减少细胞凋亡。NAD⁺显著提高耳蜗毛细胞的存活率以及减少活性氧的产生,并有效拮抗Cis引起的听力损失。转录组分析结果显示,Cis+NAD⁺组中共有204个基因上调表达、214个基因下调表达,差异表达基因显著富集于“铂类药物抗药性”和“谷胱甘肽代谢”等信号通路,并且Gstm6和Gsta2与抗氧化应激、药物代谢和细胞保护相关的关键基因表达显著上调。结论 NAD⁺在一定程度上能够拮抗Cis引起的耳毒性,其机制可能与调控谷胱甘肽代谢通路从而拮抗氧化应激损伤有关,提示了NAD⁺具有一定的治疗潜力。

关键词: 顺铂, 毛细胞, 基底膜, 烟酰胺腺嘌呤二核苷酸, 活性氧, 氧化应激, 听力障碍

Abstract: Objective The present study aims to investigate the effect and underlying mechanism of nicotinamide adenine dinucleotide(NAD⁺). The present study aims to explore the mitigating effect of NAD⁺ on cisplatin-induced ototoxicity, with a view to elucidating the underlying mechanism. Methods In vitro, HEI-OC1 cells and cochlear basement membrane cultures from 3-day-old C57BL/6J mice were utilized to evaluate the protective effects of NAD⁺ against cisplatin-induced toxicity. In vivo, the administration of cisplatin and NAD⁺ was conducted via tympanic injections in adult C57BL/6J mice, followed by auditory brainstem response testing. Transcriptome sequencing was performed to analyze differential gene expression between the cisplatin and cisplatin+ NAD⁺ groups in HEI-OC1 cells, and the quantitative polymerase chain reaction was applied to validate the key genes. Results The present study investigates the impact of NAD⁺ on the viability of HEI-OC1 cells exposed to cisplatin, with a particular focus on the phenomenon of apoptosis. The results demonstrate a significant enhancement in cell viability in the presence of NAD⁺, accompanied by a substantial reduction in apoptosis. In the cochlear basilar membrane model, NAD⁺ significantly increased hair cell survival and decreased reactive oxygen species(ROS)production. Furthermore, NAD⁺ effectively mitigated cisplatin-induced hearing loss in vivo. Transcriptome analysis revealed 204 genes to be significantly overexpressed and 214 genes to be significantly underexpressed in the cisplatin + NAD⁺ group, with differential genes notably enriched in the Platinum drug resistance and Glutathione metabolism pathways. Key genes associated with oxidative stress, drug metabolism, and cyto-protection, such as Gstm6 and Gsta2 were significantly upregulated. Conclusion NAD⁺ has been demonstrated to possess a protective effect against cisplatin-induced ototoxicity, potentially by modulating the glutathione metabolism pathway in order to counteract oxidative stress. Consequently, these findings imply that NAD⁺ has therapeutic potential in preventing cisplatin-induced hearing loss.

Key words: Cisplatin, Hair cells, Cochlear basement membrane, Nicotinamide adenine dinucleotide, Reactive oxygen species, Oxidative stress, Hearing impairment

中图分类号:

R764.43

陈铭,柯冰冰,崔雅琦,吴翠萍,陈正侬,李春燕,殷善开. NAD⁺对顺铂所致毛细胞氧化应激损伤的拮抗作用及相关基因表达调控[J]. 山东大学耳鼻喉眼学报, 2025, 39(3): 11-18.

CHEN Ming, KE Bingbing, CUI Yaqi, WU Cuiping, CHEN Zhengnong, LI Chunyan, YIN Shankai. The antagonistic effects of NAD⁺ on cisplatin-induced oxidative stress injury in hair cells and the regulation of related gene expression[J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2025, 39(3): 11-18.

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