Dissociation of HKII in retinal epithelial cells induces oxidative stress injury in the retina

Chu,Liqun; Xiao,Lin; Xu,Bing; Xu,Jingmei

doi:10.3892/ijmm.2019.4304

Dissociation of HKII in retinal epithelial cells induces oxidative stress injury in the retina

Authors:
- Liqun Chu
- Lin Xiao
- Bing Xu
- Jingmei Xu
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Affiliations: Department of Ophthalmology, Xiyuan Hospital, China Academy of Traditional Chinese Medicine, Beijing 100091, P.R. China, Department of Ophthalmology, Beijing Shijitan Hospital, CMU, Beijing 100038, P.R. China
Published online on: August 6, 2019 https://doi.org/10.3892/ijmm.2019.4304
Pages: 1377-1387
Copyright: © Chu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The retina is sensitive to injury resulting from oxidative stress (OS) due to its high oxygen consumption. Patients with retinitis pigmentosa suffer from excessive OS. N‑acetylcysteine (NAC) is used as a mucolytic agent for the clinical treatment of disorders, such as chronic bronchitis and other pulmonary diseases. The aim of the present study was to investigate the role of hexokinase 2 (HKII) in retinal OS injury. Amyloid β (Aβ)1‑40 was used to establish a cellular model of OS. Cell viability was measured with a Cell Counting Kit‑8 assay, and the apoptosis, reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) of cells were analyzed via flow cytometry with corresponding kits. The mRNA and protein levels were detected by reverse transcription‑quantitative PCR and western blot analyses, respectively. It was observed that Aβ1‑40 reduced the expression of HKII in the mitochondria of retinal pigment epithelial ARPE cells and impaired mitochondrial antioxidant functions. Additionally, knockdown of HKII promoted apoptosis, and increased ROS levels and the MMP. NAC attenuated the inhibition of mitochondrial functions induced by Aβ1‑40. The knockdown of HKII was revealed to decrease the levels of Bcl‑2, manganese superoxide dismutase (SOD) and copper‑zinc‑SOD, and increase the levels of cleaved caspase‑3, Bax and cytochrome c. The present findings suggested that the dissociation of HKII induced by OS induces apoptosis and mitochondrial damage. This study provided improved understanding of the mechanisms underlying the effects of OS on retinal epithelial cells.

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