Identification and experimental validation of ferroptosis-related gene lactotransferrin in age-related hearing loss

Chaojun Zeng; Xi Gu; Yuqing Chen; Yanchun Lin; Junying Chen; Zhifeng Chen; Chenyu Chen; Guangnan Yao; Chang Lin

doi:10.3389/fnagi.2024.1309115

Identification and experimental validation of ferroptosis-related gene lactotransferrin in age-related hearing loss

Front Aging Neurosci. 2024 Jan 11:16:1309115. doi: 10.3389/fnagi.2024.1309115. eCollection 2024.

Authors

Chaojun Zeng^#^{1

2

3}, Xi Gu^#^{1

2

3}, Yuqing Chen^#^{1

2

3}, Yanchun Lin^#^{1

2

3}, Junying Chen^#^{4

5}, Zhifeng Chen^{1

2

3}, Chenyu Chen^{1

2

3}, Guangnan Yao^{1

2

3}, Chang Lin^{1

2

3}

Affiliations

¹ Department of Otorhinolaryngology Head and Neck Surgery, The First Affiliated Hospital, Fujian Medical University, Fuzhou, China.
² Department of Otorhinolaryngology Head and Neck Surgery, National Regional Medical Center, Binhai Campus of the First Affiliated Hospital, Fujian Medical University, Fuzhou, China.
³ Fujian Institute of Otolaryngology, The First Affiliated Hospital, Fujian Medical University, Fuzhou, China.
⁴ Central Laboratory, Key Laboratory of Radiation Biology of Fujian Higher Education Institutions, The First Affiliated Hospital, Fujian Medical University, Fuzhou, China.
⁵ Central Laboratory, Key Laboratory of Radiation Biology of Fujian Higher Education Institutions, National Regional Medical Center, Binhai Campus of the First Affiliated Hospital, Fujian Medical University, Fuzhou, China.

^# Contributed equally.

Abstract

Objective: To reveal the relationship between ARHL and ferroptosis and screen ferroptosis-related genes (FRGs) in ARHL.

Methods: Bioinformatics were used to analyze the hub genes and molecular mechanism of ferroptosis in the aging cochleae. Senescence β-galactosidase staining, iron content detection, and micro malondialdehyde (MDA) assay kits were used to measure β-galactosidase activity, and expression of Fe²⁺ and MDA, respectively. Fluorescence microscope was used for immunofluorescence assay of hub genes. Western blot was used to verify the expression of hub genes in HEI-OC1 cells, cochlear explants, and cochleae of C57BL/6J mice. Data were expressed as mean ± SD of at least three independent experiments.

Results: The analysis of bioinformatics confirmed that lactotransferrin (LTF) is the hub gene and CEBPA-miR-130b-LTF network is the molecular mechanism for cochlear ferroptosis. Compared with the control group, the experiments proved that the indicators of ferroptosis, including Fe²⁺, MDA, and LTF were differentially expressed in aging HEI-OC1 cells, aging cochlear explants, and aging cochleae.

Conclusion: These results demonstrate that ferroptosis plays an important role in ARHL, and LTF is a potential therapeutic target for ARHL via regulating cochlear ferroptosis.

Keywords: age-related hearing loss; ferroptosis; hub gene; lactotransferrin; mechanism.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was funded by the Joint Funds for the Innovation of Science and Technology, Fujian Province (No. 2021Y9099), Young and Middle-aged Key Personnel Training Project of Fujian Provincial Health Commission (No. 2021GGA024), and Fujian Provincial Health Technology Project (No. 2020QNA058).