• Yonsei Medical Journal
• /
• v.59 no.9
• /
• pp.1131-1137
• /
• 2018

Purpose: Previous reports have shown that hyperglycemia-induced inhibition of transient receptor potential vanilloid sub type 4 (TRPV4), a transient receptor potential ion channel, affects the severity of hearing impairment (HI). In this study, we explored the role of TRPV4 in HI using HEI-OC1 cells exposed to high glucose (HG). Materials and Methods: HEI-OC1 cells were cultured in a HG environment (25 mM D-glucose) for 48 hours, and qRT-PCR and Western blotting were used to analyze the expression of TRPV4 at the mRNA and protein level. TRPV4 agonist (GSK1016790A) or antagonist (HC-067047) in cultured HEI-OC1 cells was used to obtain abnormal TRPV4 expression. Functional TRPV4 activity was assessed in cultured HEI-OC1 cells using the MTT assay and a cell death detection ELISA. Results: TRPV4 agonists exerted protective effects against HG-induced HI, as evidenced by increased MTT levels and inhibition of apoptosis in HEI-OC1 cells. TRPV4 overexpression significantly increased protein levels of phosphorylated p38 mitogen-activated protein kinase (p-p38 MAPK), while TRPV4 antagonists had the opposite effect. Our results indicated that TRPV4 is a hyperglycemia-related factor that can inhibit cell proliferation and promote cell apoptosis by activating the MAPK signaling pathway in HEI-OC1 cells. Conclusion: Our results show that the overexpression of TRPV4 can attenuate cell death in HEI-OC1 cells exposed to HG.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.20 no.6
• /
• pp.1524-1529
• /
• 2006

The fruits of Cornus officinalis have been used in traditional Oriental medicine for treatment of inner ear diseases, such as tinnitus and hearing loss. In the present study, we showed that the ursolic acid obtained from Corni fructus protected HEI-OC1 auditory cells from hydrogen peroxide cytotoxicity in a dose-dependent fashion. In addition, to investigate the protection mechanism of ursolic acid on hydrogen peroxide cytotoxicity toward HEI-OC1, we measured the effects of ursolic acid on lipid peroxidation and activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) in hydrogen peroxide treated cells. Ursolic acid (0.05 - 2 ${\mu}g/ml$) had protective effect against the hydrogen peroxide-induced HEI-OC1 cell damage and reduced lipid peroxidation in a dose-dependent manner. Pre-treatment with ursolic acid significantly attenuated the decrease in activities of CAT and GPX, but SOD activity was not affected by the ursolic acid or hydrogen peroxide. These results indicate that ursolic acid protects hydrogen peroxide-induced HEI-OC1 cell damage through inhibition of lipid peroxidation and induce the antioxidant enzymes CAT and GPX.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.25 no.3
• /
• pp.451-458
• /
• 2011

The medicinal mushroom Inonotus obliquus is a traditional and widely used multi-functional fungus. In this study, we have investigated whether Inonotus obliquus (Chaga mushroom) extracts exerts anti-oxidant effects on cisplatin-induced cytotoxicity in auditory cell line, HEI-OC1 cells. First of all, Chaga extracts has no harmful effects on viability of HEI-OC1 cells in the dose range of $65{\sim}125{\mu}g/m{\ell}$. Moreover, it shows cyto-protective effects on the cells treated with cisplatin-induced cytotoxicity in HEI-OC1 cells and the damage of hair cells arrays of the rat primary organ of Corti explants in the presence of cisplatin. Pretreatment with Chaga extracts inhibited the cell death, reactive oxygen species generation (ROS), lipid peroxidation induced by cisplatin. These effects were associated with the induction of antioxidant enzyme by Chaga extracts. We further investigated the effects of Chaga extracts on expression of antioxidant enzymes such as Cu, Zn superoxide dismutase (SOD 1) and Mn SOD (SOD 2) by RT-PCR. In addition, Chaga extracts shows SOD activity and SOD protein expression in cisplatin treated group induced similar to control group. Taken together, these results indicate that Chaga extracts can prevent cisplatin-induced cytotoxicity by radical-scavenging activity (SOD activity) in HEI-OC1 cells. It might be an effective as antioxidant and further studies on the chemo-preventive mechanisms of Inonotus obliquus are needed.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.21 no.4
• /
• pp.884-890
• /
• 2007

A mechanism of hair cell damage caused by noise and ototoxic agents is mediated through generation of free radicals and reactive oxygen species(ROS). It is known that most of animals have defense systems of ROS that protect against ROS, and the cochlea of animals also has ROS defense system, which appear efficient in detoxifying ROS generated under normal condition. This system includes several antioxidant enzymes such as superoxide dismutase(SOD), catalase(CAT), glutathione peroxidase (GPX), and glutathione reductase(GR). The radix of Pueraria thunbergiana(P. thunbergiana) is traditionally prescribed to attenuate the clinical manifestation of inner ear dysfunction and various clinical situations including fevers, gastrointestinal disorders, skin problems, migraine headaches, lowering cholesterol, and treating chronic alcoholism in Oriental Medicine. In the present study, to investigate the protection mechanism of ${\beta}-sitosterol$ from P. thunbergiana on cisplatin cytotoxicity toward HEI-OC1, we measured the effects of ${\beta}-sitosterol$ on activities of SOD, CAT, GPX, and GR in cisplatin treated cells. SOD, CAT, GPX, and GR activities were significantly increased in the presence of 0.001-0.1 ${\mu}g/ml$ of ${\beta}-sitosterol$ compared to the control group. These results indicate that ${\beta}-sitosterol$ protects cisplatin-induced HEI-OC1 cell damage through increasing the antioxidant enzyme system such as SOD, CAT, GPX, and GR.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.21 no.2
• /
• pp.462-467
• /
• 2007

The radix of Pueraria thunbergiana BENTHAM (Leguminosae) is traditionally prescribed to attenuate the clinical manifestations of inner ear dysfunction and various clinical situations including fever, gastrointestinal disorders, skin problems, migraine headaches, lowering cholesterol and treating chronic alcoholism in Oriental Medicine. In the present study, we examined the effect of ethanol extract of P. thunbergiana radix (EPR) on cisplatin-mediated HEI-OC1 auditory cell death. In addition, to investingate the protection mechanism of EPR on free radicals. Treatment of EPR protected cells from cisplatin and reduced lipid peroxidation in a dose-dependent manner. Furthermore, EPR demonstrated significant scavenging activity against various free radicals, including superoxide radical, hydroxyl radical, hydrogen peroxide, and DPPH radical. These results indicate that EPR protects cisplatin-induced damages of HEI-OC1 cells through inhibition of lipid peroxidation and augmenting scavenging activities against free radials.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.21 no.1
• /
• pp.214-218
• /
• 2007

The water extracts of Samultang (Samul) has been used for treatment of ischemic heart and brain damage in Oriental traditional medicine. However, little is known about the mechanism by which the water extract of Samul rescues cells from oxidative damages in cisplatin-induced ototoxicity. Cisplatin is a widely used chemotherapeutic agent that is also highly ototoxic. This study was designed to investigate the protective effects of Samul on ciplatin-induced ototoxicity in HEI-OC1 auditory cells and organ of Corti explant culture. Cisplatin markedly decreased the viability of HEI-OC1 auditory cells. However, treatment of HEI-OC1 cells with Samul significantly reduced cisplatin-induced cell death and apoptotic characteristics through reduction of intracellular peroxide generation. Cisplatin induced cytotoxicity in isolated and cultured hair cell progenitors from postnatal rat cochleae. These progenitor cells are isolated from the lesser epithelial ridge (LER, or outer spiral sulcus cell) area of pre-plated neonatal rat cochlear segments. However, Samul completely protected the morphological changes of organ of Corti and LER. Taken together, these data suggest that the protective effects of the water extracts of Samul against cisplatin may be mediated by the reduction of intracellular peroxide generation.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.19 no.5
• /
• pp.1349-1355
• /
• 2005

The steamed root of Rehmannia glutinosa has been used for treatment of inner ear diseases, such as tinnitus and hearing loss in traditional Oriental Medicine. In the present study, we investigated the effect of ethanol extract of steamed root R. glutinosa (SRG) on cisplatin cytotoxicity of HEI-OC1 auditory cells. In addition, to investigate the mechanism of SRG on cisplatin cytotoxicity, the effects of SRG on lipid peroxidation as well as scavenging activities against various free radicals were measured in cisplatin-treated cells. Treatment of SRG protected cells from cisplatin and reduced lipid peroxidation in a dose-dependent manner. Furthermore, SRG demonstrated significant scavenging activity against various free radicals, including superoxide radical, hydroxyl radical, hydrogen peroxide, and DPPH radical. These results indicate that SRG protects cisplatin-induced damages of HEI-OC1 cells through inhibition of lipid peroxidation and augmenting scavenging activities against free radials.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.19 no.6
• /
• pp.1557-1562
• /
• 2005

A mechanism of hair cell damage caused by noise and ototoxic agents is mediated through generation of free radicals and reactive oxygen species (ROS). It is known that most of animals have defense systems to protect against ROS, and the cochlea of inner ear in animals also has ROS defense systems including several antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), glutathione reductase (GR), and glutathione (GSH), which efficiently detoxifying ROS generated under normal condition. Steamed roots of Rehmannia glutinosa have been traditionally used in Oriental medicine for the treatment of auditory disease such as tinnitus, vertigo, and hearing loss as well as inflammatory diseases, hectic fever, night sweat, and headache. In the present study, we showed that the ethanol extract from steamed roots of R. giutinosa (ESRG) increased the antioxidant enzymes such as SOD, CAT, GPX, and GR activities and GSH level in HEI-OC1 auditory cells. This extract itself did not show any significant cytotoxicity up to $50{\mu}g/ml$. Our results further support the view that ESRG is promising sources of potential antioxidants. Future studies will be aimed at investigating the effects of ESRG on the regulation of cellular mechanisms and isolating and identifying the substances responsible for the regulation of antioxidant enzyme system from the plant extracts.

• Journal of Life Science
• /
• v.18 no.4
• /
• pp.542-549
• /
• 2008

Cisplatin (cis-diamminedichloroplatinum II : CDDP) is the most widely used anticancer drug against a variety of human neoplasms. However, its clinical use is limited by the onset of severe side effects, including ototoxicity and nephrotoxicity. Even though a number of evidences in cytotoxic mechanism of cisplatin have been suggested, the role of pro-inflammatory cytokines in cisplatin cytotoxicity of auditory cells has not yet been demonstrated. Herein our data clearly demonstrated that cisplatin decreased the viability of HEI-OC1 auditory cells, which was inhibited by the addition of neutralizing $anti-TNF-{\alpha}$, $anti-IL-1{\beta}$ and anti-IL-6 antibodies. Consistently, Neutralization with antibodies against pro-inflammatory cytokines ameliorated the cell death and disarrangement of cochlea hair cell layers in the rat primary cochlear explants which were treated with cisplatin. Furthermore, exogeneous supplementation with free radical scavengers, including GSH and NAC, significantly prevented the cytotoxicity of cisplatin in the rat primary cochlea explants. We also observed that $TNF-{\alpha}$ was predominantly expressed in Deiters and Hensen's cells located in hair cell zone of cisplatin-treated cochlear explants. These findings suggest that pro-inflammatory cytokines, including $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6, may play a pivotal role in the pathophysiology of hair cell damages caused by ototoxic drug cisplatin.

• Genomics & Informatics
• /
• v.5 no.3
• /
• pp.102-106
• /
• 2007

Radiofrequency (RF) radiation at the frequency of mobile phones has been not reported to induce cellular responses in in vitro and in vivo models. We exposed HEI-OC1, conditionally-immortalized mouse auditory cells, to RF radiation to characterize cellular responses to 1763 MHz RF radiation. While we could not detect any differences upon RF exposure, whole-genome expression profiling might provide the most sensitive method to find the molecular responses to RF radiation. HEI-OC1 cells were exposed to 1763 MHz RF radiation at an average specific absorption rate (SAR) of 20 W/kg for 24 hr and harvested after 5 hr of recovery (R5), alongside sham-exposed samples (S5). From the whole-genome profiles of mouse neurons, we selected 9 differentially-expressed genes between the S5 and R5 groups using information gain-based recursive feature elimination procedure. Based on support vector machine (SVM), we designed a prediction model using the 9 genes to discriminate the two groups. Our prediction model could predict the target class without any error. From these results, we developed a prediction model using biomarkers to determine the RF radiation exposure in mouse auditory cells with perfect accuracy, which may need validation in in vivo RF-exposure models.