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Drug Design, Development and Therapy Volume 13, 2019 - Issue
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Original Research

Downregulation of p66Shc can reduce oxidative stress and apoptosis in oxidative stress model of marginal cells of stria vascularis in Sprague Dawley rats

Cong Hao1 Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital Central South University, Changsha410008, Hunan, People’s Republic of China;2 Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Changsha410008, Hunan, People’s Republic of China;3 National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha 410008, Hunan, People’s Republic of China
,
Xuewen Wu1 Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital Central South University, Changsha410008, Hunan, People’s Republic of China;2 Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Changsha410008, Hunan, People’s Republic of China;3 National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha 410008, Hunan, People’s Republic of China
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Ruoyu Zhou1 Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital Central South University, Changsha410008, Hunan, People’s Republic of China;2 Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Changsha410008, Hunan, People’s Republic of China;3 National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha 410008, Hunan, People’s Republic of China
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Hao Zhang1 Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital Central South University, Changsha410008, Hunan, People’s Republic of China;2 Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Changsha410008, Hunan, People’s Republic of China
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Yulai Zhou1 Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital Central South University, Changsha410008, Hunan, People’s Republic of China;2 Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Changsha410008, Hunan, People’s Republic of China
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Xinxing Wang1 Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital Central South University, Changsha410008, Hunan, People’s Republic of China;2 Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Changsha410008, Hunan, People’s Republic of China
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Yong Feng1 Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital Central South University, Changsha410008, Hunan, People’s Republic of China;2 Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Changsha410008, Hunan, People’s Republic of China;3 National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha 410008, Hunan, People’s Republic of China
,
Lingyun Mei1 Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital Central South University, Changsha410008, Hunan, People’s Republic of China;2 Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Changsha410008, Hunan, People’s Republic of China;3 National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha 410008, Hunan, People’s Republic of China
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Chufeng He1 Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital Central South University, Changsha410008, Hunan, People’s Republic of China;2 Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Changsha410008, Hunan, People’s Republic of China;3 National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha 410008, Hunan, People’s Republic of China
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Xinzhang Cai1 Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital Central South University, Changsha410008, Hunan, People’s Republic of China;2 Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Changsha410008, Hunan, People’s Republic of China;3 National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha 410008, Hunan, People’s Republic of China
&
Lisha Wu1 Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital Central South University, Changsha410008, Hunan, People’s Republic of China;2 Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Changsha410008, Hunan, People’s Republic of China;3 National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha 410008, Hunan, People’s Republic of ChinaCorrespondence[email protected]
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Pages 3199-3206 | Published online: 10 Sep 2019
 
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Abstract

Background

p66Shc, a Src homologue and collagen homologue (Shc) adaptor protein, mediates oxidative stress signaling. The p66Shc-null mice have increased lifespan and enhanced resistance to oxidative stress. Studies have also indicated its potential role in inner ear aging, which can lead to deafness.

Objective

The aim of this study was to determine the effects of p66Shc down-regulation on the marginal cells (MCs) of the inner ear stria vascularis.

Methods

Primary MCs were isolated from neonatal rats and treated with glucose oxidase to induce oxidative stress. The cells were transduced with adenovirus expressing siRNA, and the knockdown was verified by Western blotting. The reactive oxygen species (ROS) levels and apoptosis were analyzed using the DCFH-DA probe and Annexin-V/7-AAD staining respectively. The ultrastructure of the differentially-treated cells was examined by transmission electron microscopy (TEM).

Results: The in vitro oxidative stress model was established successfully in rat MCs. Knockdown of p66Shc alleviated the high ROS levels and apoptosis in the glucose oxidase-treated cells. In addition, glucose oxidase significantly increased the number of peroxisomes in the MCs, which was decreased by p66Shc inhibition.

Conclusion

Oxidative stress increases p66Shc levels in the marginal cells of the inner ear, which aggravates ROS production and cellular injury. Blocking p66Shc expression can effectively reduce oxidative stress and protect the MCs.

Acknowledgment

This study was supported by grants from the National Nature Science Foundation of China (No. 81400468 and 81401227).

Disclosure

The authors report no conflicts of interest in this work.

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