Journal of Otolaryngology and Ophthalmology of Shandong University ›› 2025, Vol. 39 ›› Issue (3): 11-18.doi: 10.6040/j.issn.1673-3770.0.2025.054

• Shanghai Sixth Peoples Hospital Otorhinolaryngology & Head and Neck Surgery Department dedicated to “120th Anniversary of the Sixth Hospital” Commemorative Theme • Previous Articles    

The antagonistic effects of NAD+ on cisplatin-induced oxidative stress injury in hair cells and the regulation of related gene expression

CHEN Ming, KE Bingbing, CUI Yaqi, WU Cuiping, CHEN Zhengnong, LI Chunyan, YIN Shankai   

  1. Department of Otorhinolaryngology & Head and Neck Surgery, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China
  • Published:2025-06-04

PDF (PC)

9

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

CLC Number: 

  • R764.43
[1] Rancoule C, Guy JB, Vallard A, et al. 50th anniversary of cisplatin[J]. Bull Cancer, 2017, 104(2): 167-176. doi:10.1016/j.bulcan.2016.11.011
[2] Karasawa T, Steyger PS. An integrated view of cisplatin-induced nephrotoxicity and ototoxicity[J]. Toxicol Lett, 2015, 237(3): 219-227. doi:10.1016/j.toxlet.2015.06.012
[3] Hazlitt RA, Min J, Zuo J. Progress in the development of preventative drugs for cisplatin-induced hearing loss[J]. J Med Chem, 2018, 61(13): 5512-5524. doi:10.1021/acs.jmedchem.7b01653
[4] Marshak T, Steiner M, Kaminer M, et al. Prevention of cisplatin-induced hearing loss by intratympanic dexamethasone: a randomized controlled study[J]. Otolaryngol Head Neck Surg, 2014, 150(6): 983-990. doi:10.1177/0194599814524894
[5] Laurell G. Pharmacological intervention in the field of ototoxicity[J]. HNO, 2019, 67(6): 434-439. doi:10.1007/s00106-019-0663-1
[6] Gentilin E, Simoni E, Candito M, et al. Cisplatin-induced ototoxicity: updates on molecular targets[J]. Trends Mol Med, 2019, 25(12): 1123-1132. doi:10.1016/j.molmed.2019.08.002
[7] Freyer DR, Brock PR, Chang KW, et al. Prevention of cisplatin-induced ototoxicity in children and adolescents with cancer: a clinical practice guideline[J]. Lancet Child Adolesc Health, 2020, 4(2): 141-150. doi:10.1016/S2352-4642(19)30336-0
[8] Katsyuba E, Romani M, Hofer D, et al. NAD+ homeostasis in health and disease[J]. Nat Metab, 2020, 2(1): 9-31. doi:10.1038/s42255-019-0161-5
[9] Kim HJ, Oh GS, Shen A, et al. Augmentation of NAD(+)by NQO1 attenuates cisplatin-mediated hearing impairment[J]. Cell Death Dis, 2014, 5(6): e1292. doi:10.1038/cddis.2014.255
[10] Zhan T, Xiong H, Pang JQ, et al. Modulation of NAD+ biosynthesis activates SIRT1 and resists cisplatin-induced ototoxicity[J]. Toxicol Lett, 2021, 349: 115-123. doi:10.1016/j.toxlet.2021.05.013
[11] Fang J, Wu HM, Zhang JN, et al. A reduced form of nicotinamide riboside protects the cochlea against aminoglycoside-induced ototoxicity by SIRT1 activation[J]. Biomed Pharmacother, 2022, 150: 113071. doi:10.1016/j.biopha.2022.113071
[12] Hwang ES, Song SB. Possible adverse effects of high-dose nicotinamide: mechanisms and safety assessment[J]. Biomolecules, 2020, 10(5): 687. doi:10.3390/biom10050687
[13] Shen Q, Zhang SJ, Xue YZ, et al. Biological synthesis of nicotinamide mononucleotide[J]. Biotechnol Lett, 2021, 43(12): 2199-2208. doi:10.1007/s10529-021-03191-1
[14] Dehne N, Lautermann J, Petrat F, et al. Cisplatin ototoxicity: involvement of iron and enhanced formation of superoxide anion radicals[J]. Toxicol Appl Pharmacol, 2001, 174(1): 27-34. doi:10.1006/taap.2001.9171
[15] Guo XR, Bai XH, Li L, et al. Forskolin protects against cisplatin-induced ototoxicity by inhibiting apoptosis and ROS production[J]. Biomed Pharmacother, 2018, 99: 530-536. doi:10.1016/j.biopha.2018.01.080
[16] Rajman L, Chwalek K, Sinclair DA. Therapeutic potential of NAD-boosting molecules: the in vivo evidence[J]. Cell Metab, 2018, 27(3): 529-547. doi:10.1016/j.cmet.2018.02.011
[17] Brown KD, Maqsood S, Huang JY, et al. Activation of SIRT3 by the NAD+ precursor nicotinamide riboside protects from noise-induced hearing loss[J]. Cell Metab, 2014, 20(6): 1059-1068. doi:10.1016/j.cmet.2014.11.003
[18] Morita K, Maeda S, Suzuki K, et al. Paradoxical enhancement of leukemogenesis in acute myeloid leukemia with moderately attenuated RUNX1 expressions[J]. Blood Adv, 2017, 1(18): 1440-1451. doi:10.1182/bloodadvances.2017007591
[19] Ren CJ, Zhu YY, Li QQ, et al. Lespedeza bicolor Turcz. honey prevents inflammation response and inhibits ferroptosis by Nrf2/HO-1 pathway in DSS-induced human caco-2 cells[J]. Antioxidants(Basel), 2024, 13(8): 900. doi:10.3390/antiox13080900
[1] KE Bingbing, CHEN Ming, WANG Hongyang, LI Chunyan, YIN Shankai. CAMK4-mediated oxidative stress injury in auditory central neurons induced by bilirubin [J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2025, 39(3): 1-10.
[2] XU Wanjing, SUN Yuhao, ZHAO Jun. Effects of antioxidant activity of curcumin in retinal degenerative diseases [J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2025, 39(1): 146-151.
[3] ZHOU Jing, BI Xiuli, XIAO Yu, HU Jun, FU Xiaolong, YU Yafeng. Clomipramine hydrochloride protects auditory hair cells from neomycin-induced damage [J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2024, 38(4): 22-27.
[4] ZHOU Yingdong, ZHANG Mengxian, WANG Qingling, KANG Haoran, GUO Xiangdong. Progress of research of oxidative stress in the pathogenesis of presbycusis [J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2024, 38(1): 72-78.
[5] SUO Anqi, YANG Xinxin. Research progress on the relationship between mitochondrial autophagy and squamous cell carcinoma of the head and neck [J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2023, 37(3): 111-117.
[6] LI Mengting, HE Shuxi, WANG Hua. Research progress of inflammatory factors in Keratoconus [J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2023, 37(2): 151-158.
[7] SU Jie, YANG Fuyu, LI Meng, CHEN Huiru, JIANG Lisheng, WANG Lixiang. GLP-1 protected the diabetic retinopathy through induction of autophagy in rats [J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2022, 36(5): 30-34.
[8] YANG Kun, CHEN Lijuan, HE Xiaodan, LIU Zhiqi, SHA Suhua. Comparative study of ototoxicity between kanamycin and 2-hydroxypropyl-β-cyclodextrin [J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2022, 36(4): 6-11.
[9] LI Zhen, CUI Limei,SUN Yan. Advances in the role of bone morphogenetic protein-4 in inner ear development and regeneration of hair and spiral ganglion cells [J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2022, 36(2): 108-112.
[10] ZHANG Yi, WANG Wenjun,YANG Anhuai. Research progress of SIRT1 activation by resveratrol in ocular diseases [J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2022, 36(2): 151-156.
[11] FU Yihao, XU Yixuan,YAN Hong, ZHANG Jie. Recent research advances in the role of glutaredoxin in oculopathy [J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2021, 35(3): 125-130.
[12] The objective of this study was to analyze the effects and possible regulatory mechanisms that Notch receptors could have on cisplatin resistance, observed in nasopharyngeal carcinoma. MethodsWestern blot analysis was used for the detection of Notch receptor expression in nasopharyngeal carcinoma cells and cisplatin-resistant nasopharyngeal carcinoma cells(5-8F, 5-8F/CDDP). Flow cytometry was used to investigate how the combined treatment with 10 μM CDDP and different concentrations of γ-secretase inhibitor(DAPT)could affect apoptosis in 5-8F/CDDP cells. Flow cytometry was also used for the detection of cell cycle stages in DAPT-treated 5-8F / CDDP cells. Finally, western blotting was also used for the detection of drug resistance-related protein expression. All experiments were followed by statistical data analysis. ResultsWe observed significantly higher Notch1 and Notch4 receptor expression in 5-8F/CDDP cells than in 5-8F cells(P=0.003, P=0.004). Furthermore, we described that Notch signaling was inhibited by DAPT in 5-8F/CDDP cells, followed by a significant increase in the apoptosis rate and decrease in cell proliferation, induced by cisplatin in a dose-dependent manner(P<0.05). Moreover, after inhibiting the Notch signaling pathway in 5-8F/CDDP cells, DAPT treatment significantly decreased the expression of Cyclin E and CDK-2, proteins involved in cell cycle regulation, and contributed to blocking the cells in the G1/S phase(P<0.05). At the same time, the expression levels of both the EMT-related protein Slug and the DNA excision repair protein ERCC1 significantly decreased, while that of E-Cadherin was up-regulated. ConclusionUp-regulated expression of Notch1 and Notch4 receptors is associated with cisplatin resistance in nasopharyngeal carcinoma cells. The inactivation of the Notch signaling pathway might thus have the potential to enhance the efficiency of cisplatin chemotherapy in drug-resistant nasopharyngeal carcinoma cells by inhibiting EMT rather than blocking the G1/S cell cycle phase.. Cisplatin resistance in nasopharyngeal carcinoma cells is affected by Notch receptors through the regulation of epithelial-mesenchymal transition rather than cell cycle controlHAN Jibo, ZOU You, YANG Rui, TAO Zezhang Department of Otolaryngology-Head and Neck Surgery, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei, ChinaAbstract:Objective〓 [J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2020, 34(4): 105-110.
[13] SONG Fan, HUANG Weijun, XU Huajun, GUAN Jian, YI Hongliang. Relationship between carotid elasticity and oxidative stress in patients with obstructive sleep apnea syndrome [J]. Journal of Otolaryngology and Ophthalmology of Shandong University, 2019, 33(4): 99-104.
[14] ZHANG Zhuan, LIU Tao, BAI Zhili, ZHOU Changming. Evolution of oxidative stress in the pathogenesis and treatment of noise-induced hearing loss. [J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2017, 31(5): 101-103.
[15] . Effects of rosiglitazone on oxidative stress and cognitive function in mice exposed to intermittent hypoxia. [J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2017, 31(1): 45-49.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Website Copyright: Editorial Office of Journal of Otolaryngology and Ophthalmology of Shandong University
Address: No.27 Shanda South Road, Jinan, Shandong, China. Post code: 250100 Tel: +86-0531-88366912 Email: ebhxb@sdu.edu.cn Supported by Beijing Magtech Co.Ltd