Journal of Otolaryngology and Ophthalmology of Shandong University ›› 2025, Vol. 39 ›› Issue (3): 1-10.doi: 10.6040/j.issn.1673-3770.0.2024.509

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

CAMK4-mediated oxidative stress injury in auditory central neurons induced by bilirubin

KE Bingbing1, CHEN Ming1, WANG Hongyang2, LI Chunyan1, YIN Shankai1   

  1. 1. Shanghai Key Laboratory of Sleep Disordered Breathing, Department of Otolaryngology-Head and Neck Surgery, Otolaryngology Institute of Shanghai JiaoTong University, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200233, China2. Department of Audiology and Vestibular Medicine, College of Otolaryngology Head and Neck Surgery, Chinese PLA General Hospital, Chinese PLA Medical School, National Clinical Research Centerfor Otolaryngologic Diseases, Key Labof Hearing Science, Ministry of Education, Beijing Key Lab of Hearing Impairment for Prevention and Treatment, Beijing 100853, China
  • Published:2025-06-04

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Abstract: Objective The present study aims to investigate the role of calcium/calmodulin-dependent protein kinase 4(CAMK4)in bilirubin-mediated auditory central neurotoxicity. Methods The binding ability between CAMK4 and bilirubin was assessed using molecular docking and micro-scale thermophoresis(MST). The expression of CAMK4 in the cochlear nucleus was detected by immunofluorescence staining on paraffin sections. Primary cochlear nucleus neurons were cultured in vitro and treated with bilirubin(25 μmol/L)or bilirubin combined with the CAMK4 inhibitor KN-93(1 μmol/L). The neuronal ROS levels, mitochondrial membrane potential changes, Annexin V/PI staining, and live/dead cell staining were then evaluated. Results The molecular docking analysis revealed that the binding energy between bilirubin and CAMK4 was -9.71 kcal/mol, and the MST analysis gave a dissociation constant of(1.294 4±1.080 3)μmol/L, suggesting a strong binding affinity between the two. In the cochlear nucleus, CAMK4 was predominantly expressed in neurons. In vitro experiments demonstrated that the CAMK4 inhibitor KN-93 significantly suppressed bilirubin-induced ROS accumulation, mitochondrial membrane potential decline, late-stage apoptosis, and reduced cell survival in cochlear nucleus neurons. Conclusion CAMK4 has been demonstrated to play a crucial role in bilirubin-mediated oxidative stress injury in auditory central neurons.

Key words: Calcium/calmodulin-dependent protein kinase IV, bilirubin, neurotoxicity, oxidative stress

CLC Number: 

  • R764.4
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