Catalpol Protects ARPE-19 Cells against Oxidative Stress

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Article Catalpol Protects ARPE-19 Cells against Oxidative Stress via Activation of the Keap1/Nrf2/ARE Pathway Longtai You, Hulinyue Peng, Jing Liu, Mengru Cai, Huimin Wu, Zhiqin Zhang, Jie Bai, Yu Yao, Xiaoxv Dong *, Xingbin Yin * and Jian Ni * Citation: You, L.; Peng, H.; Liu, J.; Cai, M.; Wu, H.; Zhang, Z.; Bai, J.; Yao, Y.; Dong, X.; Yin, X.; et al. Catalpol Protects ARPE-19 Cells against Oxidative Stress via Activation of the Keap1/Nrf2/ARE Pathway. Cells 2021, 10, 2635. https:// doi.org/10.3390/cells10102635 Academic Editor: Anna-Liisa Nieminen Received: 15 June 2021 Accepted: 30 September 2021 Published: 2 October 2021 Publisher’s Note: MDPI stays neu- tral with regard to jurisdictional claims in published maps and institu- tional affiliations. Copyright: © 2021 by the authors. Li- censee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and con- ditions of the Creative Commons At- tribution (CC BY) license (https://cre- ativecommons.org/licenses/by/4.0/). School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China; 20190941307@bucm.edu.cn (L.Y.); 20200935154@bucm.edu.cn (H.P.); 20210941441@bucm.edu.cn (J.L.); cmrtcm@bucm.edu.cn (M.C.); 20190935128@bucm.edu.cn (H.W.); 20190935126@bucm.edu.cn (Z.Z.); 20190935127@bucm.edu.cn (J.B.); 20190935142@bucm.edu.cn (Y.Y.) * Correspondence: 201801020@bucm.edu.cn (X.D.); yxbtcm@bucm.edu.cn (X.Y.); 602054@bucm.edu.cn (J.N.) Abstract: Oxidative damage to retinal pigment epithelial (RPE) has been identified as one of the major regulatory factors in the pathogenesis of age-related macular degeneration (AMD). Catalpol is an iridoid glucoside compound that has been found to possess potential antioxidant activity. In the present study, we aimed to investigate the protective effect of catalpol on RPE cells under oxi- dative stress and to elucidate the potential molecular mechanism involved. We found that catalpol significantly attenuated hydrogen peroxide (H2O2)-induced cytotoxicity, G0/G1 phase cell cycle ar- rest, and apoptosis in RPE cells. The overproduction of reactive oxygen species (ROS) and malondialdehyde (MDA) stimulated by oxidative stress and the corresponding reductions in anti- oxidant glutathione (GSH) and superoxide dismutase (SOD) levels were largely reversed by catal- pol pretreatment. Moreover, catalpol pretreatment markedly activated the expression of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and its downstream antioxidant enzymes, catalase (CAT), heme oxygenase-1 (HO-1), and NADPH dehydrogenase (NQO1). It also increased the expression levels of cyclin E Bcl-2, cyclin A, and cyclin-dependent kinase 2 (CDK2) and decreased the expres- sion levels of Bax, Fas, cleaved PARP, p-p53, and p21 cleaved caspase-3, 8, and 9. The oxidative stress-induced formation of the Keap1/Nrf2 complex in the cytoplasm was significantly blocked by catalpol pretreatment. These results indicate that catalpol protected RPE cells from oxidative stress through a mechanism involving the activation of the Keap1/Nrf2/ARE pathways and the inactiva- tion of oxidative stress-mediated pathways of apoptosis. Keywords: catalpol; oxidative stress; Nrf2; apoptosis; cell cycle arrest; age-related macular degeneration 1. Introduction Age-related macular degeneration (AMD) is an irreversible visual impairment asso- ciated disease prevalent in the elderly population around the world. The clinical symp- toms of AMD are the visual distortion of straight lines, the presence of central dark spots, and decreased central vision [1–5]. Oxidative stress-induced retinal pigment epithelial (RPE) cell dysfunction and apoptosis have been identified as crucial factors involved in the pathogenesis of early AMD [6]. RPE cells are of highly active monolayer cells on the Bruch’s membrane between the neurosensory retina and the vascular choroid, and they play a very important role in maintaining the functions of retina and photoreceptors [7]. Therefore, RPE cells are vulnerable to the oxidative stress damage caused by stimulation from external factors (such as smoke and ultraviolet radiation) or high levels of intracel- lular reactive oxygen species (ROS) [8]. Oxidative stress damage leads to mitochondrial dysfunction and the production of abnormal lipid molecules in RPE cells. These products accumulate inside the Bruch’s membrane, leading to the formation of drusen which is one Cells 2021, 10, 2635. https://doi.org/10.3390/cells10102635 www.mdpi.com/journal/cells

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