Involvement of NADPH oxidases in alkali burn-induced corneal injury

Gu,Xue-Jun; Liu,Xian; Chen,Ying-Ying; Zhao,Yao; Xu,Man; Han,Xiao-Jian; Liu,Qiu-Ping; Yi,Jing-Lin; Li,Jing-Ming

doi:10.3892/ijmm.2016.2594

Involvement of NADPH oxidases in alkali burn-induced corneal injury

Authors:
- Xue-Jun Gu
- Xian Liu
- Ying-Ying Chen
- Yao Zhao
- Man Xu
- Xiao-Jian Han
- Qiu-Ping Liu
- Jing-Lin Yi
- Jing-Ming Li
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Affiliations: Affiliated Eye Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China
Published online on: May 17, 2016 https://doi.org/10.3892/ijmm.2016.2594
Pages: 75-82
Copyright: © Gu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Chemical burns are a major cause of corneal injury. Oxidative stress, inflammatory responses and neovascularization after the chemical burn aggravate corneal damage, and lead to loss of vision. Although NADPH oxidases (Noxs) play a crucial role in the production of reactive oxygen species (ROS), the role of Noxs in chemical burn-induced corneal injury remains to be elucidated. In the present study, the transcription and expression of Noxs in corneas were examined by RT-qPCR, western blot analysis and immunofluorescence staining. It was found that alkali burns markedly upregulated the transcription and expression of Nox2 and Nox4 in human or mouse corneas. The inhibition of Noxs by diphenyleneiodonium (DPI) or apocynin (Apo) effectively attenuated alkali burn-induced ROS production and decreased 3-nitrotyrosine (3-NT) protein levels in the corneas. In addition, Noxs/CD11b double‑immunofluorescence staining indicated that Nox2 and Nox4 were partially co-localized with CD11b. DPI or Apo prevented the infiltration of CD11b-positive inflammatory cells, and inhibited the transcription of inflammatory cytokines following alkali burn-induced corneal injury. In our mouse model of alkali burn-induced corneal injury, corneal neovascularization (CNV) occurred on day 3, and it affected 50% of the whole area of the cornea on day 7, and on day 14, CNV coverage of the cornea reached maximum levels. DPI or Apo effectively attenuated alkali burn‑induced CNV and decreased the mRNA levels of angiogenic factors, including vascular endothelial growth factor (VEGF), VEGF receptors and matrix metalloproteinases (MMPs). Taken together, our data indicate that Noxs play a role in alkali burn-induced corneal injury by regulating oxidative stress, inflammatory responses and CNV, and we thus suggest that Noxs are a potential therapeutic target in the future treatment of chemical-induced corneal injury.

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