Hiroaki Shimogori

Geranylgeranylacetone, a heat shock protein inducer, prevents acoustic injury in the guinea pig

Geranylgeranylacetone (GGA) used widely as anti-ulcer agent is accepted as an inducer of the heat shock proteins (Hsps) at gastric mucosa, liver, heart, and brain. However, there have been no reports that GGA could induce Hsps in the cochlea leading up to the oto-protection. The purpose of the present study was to investigate whether single oral dose of GGA could induce Hsps at cochlea and oral administration had protective effect to the cochlea against noise trauma. We used Hartley guinea pigs and investigated the expression of Hsp70, 40, and 27 in cochlea by Western blot analysis. To evaluate cochlear function, we assessed thresholds of the auditory brain stem response (ABR). For histological assessment, we observed the sensory epithelium using surface preparation technique. GGA (600 mg/kg) or vehicle was given orally to animals. Western blot analysis showed that the expressions of Hsp 70, 40, and 27 were increased 24-48 h after administration of single dose of GGA, whereas there was less expression in the animals given vehicle. In the animals given GGA once a day for a week before sound exposure (130 dB SPL octave band noise with a center frequency of 4 kHz) for 3 h, their ABR threshold elevations were lowered significantly. In addition, significantly fewer defects were observed on outer hair cells of organ of Corti in the animals treated by GGA than those of the animals without GGA. This result shows that pretreatment by GGA have a potential to prevent cochlea damage against the intense noise.

Attenuation of progressive hearing loss in a model of age-related hearing loss by a heat shock protein inducer, geranylgeranylacetone.

Mechanisms of age-related hearing loss (ARHL) have not been elucidated as aging processes are extremely complex. Although oxidative stress and apoptotic cell death are involved in progression of ARHL, number of trial to treat ARHL is limited. Heat shock response is characterized by induction of heat shock proteins (HSPs) in response to stresses such as heat shock, which diminishes during aging. HSPs act as molecular chaperones, and some HSPs also inhibit apoptotic pathways. Here, we examined age-related expression of HSPs in the cochlea of ARHL model DBA/2J mice and control CBA/N mice. Western blot assay revealed that CBA/N mice showed constant expression of Hsp70 and Hsp110 with age, but not in DBA/2J mice. The result suggests that pharmacological upregulation of HSPs might attenuate ARHL. We administered DBA/2J mice with food containing geranylgeranylacetone (GGA) that induces HSPs in the cochlea, and found that its administration suppresses ARHL examined by ABR test and histological examination though protection is specific for the apical part of the cochlea. These results demonstrate that dietary supplementation of GGA could be an effective therapeutic strategy for treatment of ARHL.

Protective effect of edaravone against streptomycin-induced vestibulotoxicity in the guinea pig

This study investigated alleviation of streptomycin-induced vestibulotoxicity by edaravone in guinea pigs. Edaravone, a free radical scavenger, has potent free radical quenching action and is used in clinical practice to treat cerebral infarction. Streptomycin was administered to the inner ear by osmotic pump for 24 h, and edaravone (n=8) or saline (n=6) was intraperitoneally injected once a day for 7 days. We observed horizontal vestibulo-ocular reflex as a marker of postoperative vestibular function. Animals injected with saline showed statistically smaller gains than those injected with edaravone. These results suggest that edaravone suppresses streptomycin-induced vestibulotoxicity.

Inner Ear Changes With Intracochlear Gentamicin Administration in Guinea Pigs

Objectives/Hypothesis Transtympanic administration of gentamicin is reported to be a useful treatment for vertigo in such conditions as Menieres disease, and determining appropriate clinical dosage of gentamicin is difficult. The authors examined the relation between gentamicin dosages and inner ear function in guinea pigs.

Geranylgeranylacetone suppresses noise-induced expression of proinflammatory cytokines in the cochlea

OBJECTIVE Heat shock transcription factor 1 (HSF1) is a master regulator of heat shock response, and also inhibits expression of inflammatory cytokines directly or indirectly. Here, we examined effects of HSF1 activation on the expression of proinflammatory cytokines in mouse cochlea after exposure to noise. METHODS Male CBA/N mice with normal Preyers reflex were exposed to intense noise for 3h. Three hours after noise exposure, bilateral cochleae were removed and expression of major inflammatory cytokines was examined. RESULTS We found that interleukin-6 (IL-6) and interleukin-1β (IL-1β) expression increased significantly after noise exposure, and the expression was suppressed significantly in mice administered with geranylgeranylacetone (GGA), which activates HSF1. Seven days after noise exposure, thresholds for auditory brainstem response were elevated, and GGA administration significantly suppressed this elevation. CONCLUSION These results suggest that HSF1-mediated suppression of proinflammatory cytokines in the cochlea by GGA administration could be an important means of inner ear protection.

Cell Proliferation in Spiral Ligament of Mouse Cochlea Damaged By Dihydrostreptomycin Sulfate

Cell proliferation of the spiral ligament in the normal and drug-induced damaged mice cochleae was investigated using the mitotic tracer bromodcoxyuridine (BrdU). Only a few nuclei labelled by BrdU were seen in the spiral ligament of the control mouse. However, many BrdU labelled nuclei in the spiral ligament in the cochlea damaged by dihydrostreptomycin sulfate were found. The expression of fibroblast growth factor receptor and connexin 43 was detected in the spiral ligament where BrdU labelled cells were found. These results suggest that cell proliferation in the spiral ligament may occur after the drug-induced damage, and this process is probably related to the recovery of cochlear function.

Effects of carbonic anhydrase inhibitor on the otolithic organs of developing chick embryos

Carbonic anhydrase appears to be involved in the process of otoconial formation. The purpose of this investigation was to observe the morphologic change in the surface structure of the otolithic organ in developing chick embryos after injection of the carbonic anhydrase inhibitor, acetazolamide. Acetazolamide (1.5, 3, or 6 mg/0.06 mL/egg) was injected into the yolk sac of the embryo of the fifth day of incubation. Embryo specimens were collected on the 11th, 13th, and 18th days of incubation. The chicks were killed on the third day posthatching, and the surfaces of the otolithic organs were observed under a scanning electron microscope. A marked disturbance in otoconial formation was noted in both utricle and saccule, marked by a decrease or absence of otoconia. A widely exposed meshwork structure of otolithic membrane was observed, with sensory cilia penetrating the meshwork small holes in many instances. There were also several otoconial abnormalities, such as the appearance of only a single giant otoconium, or from several to dozens of giant otoconia, and rough, spongy-surfaced global substances entirely covering the maculae. Clearly, carbonic anhydrase inhibitor (acetazolamide) injected into the yolk sac of developing chick embryos alters and inhibits normal otoconial morphogenesis.

A role of glucocorticoid receptors in the guinea pig vestibular system.

To investigate glucocorticoid receptor (GR) function in the vestibular periphery, GR antagonist RU38486 was administered to the guinea pig inner ear by osmotic pump, and we observed post-rotatory nystagmus (PRN) changes as a marker of vestibular function. Ten days after treatment, RU38486 (1 mM) resulted in ipsilateral vestibule hyperexcitability in response to rotation stimulation. This effect was dose-dependent. These data indicate that steroid hormones may play an important role in maintaining vestibular function.

Effect of Edaravone on Streptomycin-Induced Vestibulotoxicity in the Guinea Pig

Objectives/Hypothesis: The effect of topical administration of edaravone to the inner ear was investigated in guinea pigs with streptomycin‐induced vestibulotoxicity.

Cochlear Administration of Adenosine Triphosphate Facilitates Recovery from Acoustic Trauma (Temporary Threshold Shift)

Background: Adenosine triphosphate (ATP) has often been used in the treatment of acoustic trauma although evidence supporting its clinical use was lacking. The aim of this study was to evaluate the chronic effects of ATP on acoustic trauma in guinea pigs. Methods: We infused ATP into the perilymph of the guinea pig cochlea concurrently with intense noise exposure to investigate the effect of ATP on the process of recovery after acoustic trauma. We assessed auditory brainstem response (ABR) thresholds to evaluate cochlear function. Results: After noise exposure (120 dB SPL, 5 h), ABR thresholds showed an increase of approximately 50 dB SPL that returned to normal after 14 days. Cochlear function in ATP-treated ears recovered more quickly than in control ears. The effect of ATP was inhibited by the administration of the ATP receptor antagonist: pyridoxal- phosphate-6-azophenyl-2′,4′-disulfonic acid. Conclusion: These results suggest that ATP mitigates the effects of noise trauma through the ATP receptor.