Pharmacological Inhibition of Cochlear Mitochondrial Respiratory Chain Induces Secondary Inflammation in the Lateral Wall: A Potential Therapeutic Target for Sensorineural Hearing Loss

Masato Fujioka,Yasuhide Okamoto,Tatsuo Matsunaga,Kaoru Ogawa,Hideyuki Okano,Hirotaka James Okano,Seiichi Shinden,Partha Mukhopadhyay

doi:10.1371/journal.pone.0090089

Masato Fujioka, Yasuhide Okamoto + Show 6 more

Open Access

https://doi.org/10.1371/journal.pone.0090089

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Abstract

Cochlear lateral wall has recently been reported as a common site of inflammation, yet precise molecular mechanisms of the inflammatory responses remain elucidated. The present study examined the inflammatory responses in the lateral wall following acute mitochondrial dysfunction induced by a mitochondrial toxin, 3-nitropropionic acid (3-NP). Reverse-transcription (RT)-PCR revealed increases in the expression of the proinflammatory cytokines interleukin (IL)-1β and IL-6. Immunohistochemistry showed an increase in the number of activated cochlear macrophages in the lateral wall, which were in close proximity to IL-6-expressing cells. A genome-wide DNA microarray analysis of the lateral wall revealed that 35% and 60% of the genes showing >2-fold upregulation at 1 d and 3 d post-3-NP administration, respectively, were inflammatory genes, including CC- and CXC-type chemokine genes. High expression of CCL-1, 2, and 3 at 1 d, and of CCL-1, 2, 3, 4, and 5, CCR-2 and 5, and CX3CR1 at 3 d post-3-NP administration, coupled with no change in the level of CX3CL1 expression suggested that macrophages and monocytes may be involved in the inflammatory response to 3-NP-mediated injury. Quantitative (q)RT-PCR showed a transient induction of IL-1β and IL-6 expression within 24 h of 3-NP-mediated injury, followed by sustained expression of the chemoattractants, CCL-2, 4 and 5, up until 7 d after injury. The expression of CCL-2 and IL-6 was higher in animals showing permanent hearing impairment than in those showing temporary hearing impairment, suggesting that these inflammatory responses may be detrimental to hearing recovery. The present findings suggest that acute mitochondrial dysfunction induces secondary inflammatory responses in the lateral wall of the cochlear and that the IL-6/CCL-2 inflammatory pathway is involved in monocyte activation. Therefore, these secondary inflammatory responses may be a potential post-insult therapeutic target for treatments aimed at preventing the damage caused by acute mitochondrial dysfunction in the cochlear lateral wall.

Highlights

Mitochondrial dysfunction in the cochlea is a well-known cause of sensorineural hearing loss
Because the damaged spiral ligament is a common site of inflammation induced by various types of insult, including surgical stress, noise exposure, and immune-mediated treatments [6,9,18,21], the present study examined the inflammatory responses following acute mitochondrial dysfunction in the cochlea using both TTS and PTS models induced by treatment with 3-nitropropionic acid (3-NP)
We observed a temporal upregulation in IL-6 expression 1 d after 3-NP treatment; no such increase was noted in the controls

Summary

Introduction

Mitochondrial dysfunction in the cochlea is a well-known cause of sensorineural hearing loss. Mutations in mitochondrial DNA cause both syndromic and nonsyndromic deafness, and the inner ear is considered highly susceptible to mitochondrial dysfunction [5,11,19]. We established a novel rat model of acute mitochondrial dysfunction in the cochlea by applying 3-nitropropionic acid (3-NP) directly to the round window membrane [10]. Because local ATP deprivation in the inner ear results from inhibition of inner ear mitochondrial function. This model replicates the etiology of inner ear energy failure caused by ATP deprivation due to inner ear ischemia. It has been reported that inner ear ischemia that results from occlusion of the anterior inferior cerebellar artery causes sensorineural hearing loss [14,24]

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