Retinoprotective effect of donepezil in diabetic mice involves mitigation of excitotoxicity and activation of PI3K/mTOR/BCl2 pathway

• Published in: Life sciences (1973) 2020-12, Vol.262, p.118467-14, Article 118467
• Main Authors: Zaitone, Sawsan A., Alshaman, Reem, Alattar, Abdullah, Elsherbiny, Nehal M., Abogresha, Noha M., El-Kherbetawy, Mohammed K., Elaskary, Abdelhakeem A., Hashish, Abdullah A., Rashed, Laila A., Ahmed, Eman
• Format: Article
• Language: English
• Subjects: 1-Phosphatidylinositol 3-kinase; Acetylcholine; Apoptosis; B-cell lymphoma; Beta cells; Caspase-3; Cytokines; Diabetes; Diabetes mellitus; Diabetic retinopathy; Donepezil; Excitotoxicity; Glutamate; Glutamic acid receptors; Histopathology; Immunohistochemistry; Inflammation; Kinases; Lymphoma; Mitigation; Mouse; N-Methyl-D-aspartic acid receptors; Neuroprotection; Oxidation; Phosphorylation; PI3K/mTOR/BCl2; Proteins; Retina; Retinopathy; Thickness; TOR protein
• ISSN: 0024-3205; 1879-0631
• DOI: 10.1016/j.lfs.2020.118467

Donepezil (DNPZ) has shown neuroprotective effect in many disorders. The current study tested the putative retinoprotection provided by donepezil in mouse diabetic retinopathy. Swiss albino mice were allocated to, 1] saline control, 2] diabetic, 3&4] diabetic+DNPZ (1 or 4 mg/kg). After induction of diabetes, mice were maintained for 8 weeks then DNPZ therapy was launched for 28 days. Retinas were isolated and used for histopathology and immunohistochemistry for caspase 3 and the anti-apoptotic protein, B-cell lymphoma 2 (BCl2). Retinas were examined for glutamate, acetylcholine and oxidation markers. Western blot analysis measured inflammatory cytokines, N-methyl-d-aspartate receptors (NMDARs), phosphorylated and total phosphatidylinositol-3 kinase and mTOR, BCl2 and cleaved caspase 3. Significant histopathological changes and decreased thickness were found in diabetic retinas (125.52 ± 2.85 vs. 157.15 ± 7.55 in the saline group). In addition, retinal glutamate (2.39-fold), inflammatory cytokines and NMDARs proteins (4.9-fold) were higher in the diabetic retinas. Western blot analysis revealed low ratio of phosphorylated/total PI3K (0.21 ± 0.043 vs. 1 ± 0.005) and mTOR (0.18 ± 0.04 vs. 1 ± 0.005), low BCl2 (0.28 ± 0.06 vs. 1 ± 0.005) and upregulated cleaved caspase 3 (5.18 ± 1.27 vs. 1 ± 0.05 in the saline group) versus the saline control. DNPZ ameliorated the histopathologic manifestations and to prevent the decrease in retinal thickness. DNPZ (4 mg/kg) improved phosphorylation of PI3K (0.76 ± 0.12 vs. 0.21 ± 0.04) and mTOR (0.59 ± 0.09 vs. 0.18 ± 0.04) and increased BCl2 (0.75 ± 0.08 vs. 0.28 ± 0.06) versus the diabetic control group. This study explained the retinoprotective effect of DNPZ in mouse diabetic retinopathy and highlighted that mitigation of excitotoxicity, improving phosphorylation of PI3K/mTOR and increasing BCl2 contribute to this effect.