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Apocynin ameliorates NADPH oxidase 4 (NOX4) induced oxidative damage in the hypoxic human retinal Müller cells and diabetic rat retina

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Abstract

NADPH oxidase (NOX) is a main producers of reactive oxygen species (ROS) that may contribute to the early pathogenesis of diabetic retinopathy (DR). ROS has harmful effects on endogenous neuro-survival factors brain-derived neurotrophic factor (BDNF) and sirtuin 1 (SIRT1) are necessary for the growth and survival of the retina. The role of NOX isoforms NOX4 in triggering ROS in DR is not clear. Here we determine the protective effects of a plant-derived NOX inhibitor apocynin (APO) on NOX4-induced ROS production which may contribute to the depletion of survival factors BDNF/SIRT1 or cell death in the diabetic retinas. Human retinal Müller glial cells (MGCs) were treated with hypoxia mimetic agent cobalt chloride (CoCl2) in the absence or presence of APO. Molecular analysis demonstrates that NOX4 is upregulated in CoCl2-treated MGCs and in the diabetic retinas. Increased NOX4 was accompanied by the downregulation of BDNF/SIRT1 expression or in the activation of apoptotic marker caspase-3. Whereas, APO treatment downregulates NOX4 and subsequently upregulates BDNF/SIRT1 or alleviate caspase-3 expression. Accordingly, in the diabetic retina we found a positive correlation in NOX4 vs ROS (p = 0.025; R2 = 0.488) and caspase-3 vs ROS (p = 0.04; R2 = 0.428); whereas a negative correlation in BDNF vs ROS (p = 0.009; R2 = 0.596) and SIRT1 vs ROS (p = 0.0003; R2 = 0.817) respectively. Taken together, NOX4-derived ROS could be a main contributor in downregulating BDNF/SIRT1 expression or in the activation of caspase-3. Whereas, APO treatment may minimize the deleterious effects occurring due to hyperglycemia and/or diabetic mimic hypoxic condition in early pathogenesis of DR.

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Acknowledgement

The authors thank for funding by Dr. Nasser Al-Rasheed Research Chair in Ophthalmology (Abu El-Asrar A.M.), King Saud University, KSA.

Funding

Sources of Support: This work was supported by Dr. Nasser Al-Rasheed Research Chair in Ophthalmology (Abu El-Asrar A.M.), King Saud University, K.S.A.

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  1. Ajmal Ahmad and Mohd Imtiaz Nawaz contributed equally to this work.

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  1. Department of Ophthalmology, College of Medicine, King Saud University, Riyadh, Saudi Arabia

    Ajmal Ahmad, Mohd Imtiaz Nawaz, Mohammad Mairaj Siddiquei & Ahmed M. Abu El-Asrar

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  1. Ajmal Ahmad
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AJ and MIN contributed to the conception, design of the study, and drafted the manuscript. MMS, AJ, and MIN performed the experiments and conducted the data analysis. AMAE edited the manuscript and provided comments. AJ is responsible for the integrity and accuracy of the work, as well have full access to all the data in the study. All authors read and revised the manuscript for intellectual content and approved the submitted version.

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Correspondence to Ajmal Ahmad.

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Ahmad, A., Nawaz, M.I., Siddiquei, M.M. et al. Apocynin ameliorates NADPH oxidase 4 (NOX4) induced oxidative damage in the hypoxic human retinal Müller cells and diabetic rat retina. Mol Cell Biochem 476, 2099–2109 (2021). https://doi.org/10.1007/s11010-021-04071-y

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