ANTIOXIDANT AND MODULATION OF APOPTOSIS-RELATED PATHWAYS OF MAKIANG FRUIT EXTRACT IN ARPE-19 CELLS EXPOSED TO GLUCOSE-INDUCED OXIDATIVE STRESS
DOI:
https://doi.org/10.69598/tbps.21.2.215-228%20Keywords:
Makiang Fruit, diabetic retinopathy, human retinal pigment epithelial ARPE-19 cells, oxidative stress, anthocyanin, antioxidant defenseAbstract
Diabetic retinopathy (DR) is a leading cause of visual loss resulting from microvascular complications of diabetes mellitus. In DR, retinal pigment epithelial (RPE) cells are exposed to elevated blood sugar levels for prolonged periods, leading to oxidative stress and apoptosis. Makiang fruit (Cleistocalyx nervosum var. paniala) is rich in anthocyanins (ACN), which possess antioxidant and anti-apoptotic properties. However, its protective effects on RPE cells against glucose-induced oxidative stress have not been studied. This study aimed to examine the effects of Makiang fruit extract on human retinal pigment epithelial (ARPE-19) cells exposed to glucose-induced oxidative stress. Makiang fruits were extracted with 95% ethanol (MKE95). The results showed that MKE95 exhibited antioxidant capacity as measured by the DPPH, ORAC, and FRAP assays. For the cell-based experiments, ARPE-19 cells were co-treated with MKE95 (50–200 μg/mL) and 0.5% DMSO in DMEM/F-12 (5.5 mM glucose), with D-glucose adjusted to 30 mM. At the highest concentration, MKE95 significantly (p < 0.05) improved cell viability by 15.4%, reduced intracellular ROS by 37.8%, and decreased caspase-9 and -3 activities (1.66- and 1.64-fold, respectively) compared to the glucose-treated group. It also enhanced antioxidant defenses, increasing SOD, CAT, GPx activities, and GSH levels by 22.4%, 26.3%, 24.7%, and 24.4%, respectively, in a dose-dependent manner. However, these findings are derived from an in vitro model. Although Makiang extract demonstrates protective effects against glucose-induced oxidative stress in RPE cells, further in vivo and clinical studies are necessary to confirm its potential role in preventing diabetic retinopathy.
References
Callan A, Heckman J, Tah G, Lopez S, Valdez L, Tsin A. VEGF in diabetic retinopathy and age-related macular degeneration. Int J Mol Sci. 2025;26(11):4992.
Hayati A, Abdol Homayuni MR, Sadeghi R, Asadigandomani H, Dashtkoohi M, Eslami S, et al. Advancing diabetic retinopathy screening: A systematic review of artificial intelligence and optical coherence tomography angiography innovations. Diagnostics (Basel). 2025;15(6):737.
Force UPST, Davidson KW, Barry MJ, Mangione CM, Cabana M, Caughey AB, et al. Screening for prediabetes and type 2 diabetes: US Preventive Services Task Force recommendation statement. JAMA. 2021;326(8):736-43.
Mohammed MR, Fathi SA. Global prevalence of diabetic retinopathy over the last decade (2015–2025): A scoping review. medRxiv [Preprint]. 2025:2025.07.24.25332019.
Benhamza M, Dahlui M, Said MA. Determining direct, indirect healthcare and social costs for diabetic retinopathy management: a systematic review. BMC Ophthalmol. 2024;24(1):424.
Kaštelan S, Konjevoda S, Sarić A, Urlić I, Lovrić I, Čanović S, et al. Resveratrol as a novel therapeutic approach for diabetic retinopathy: Molecular mechanisms, clinical potential, and future challenges. Molecules. 2025;30(15):3262.
Gong X, Draper C, Allison G, Marisiddaiah R, Rubin L. Effects of the macular carotenoid lutein in human retinal pigment epithelial cells. Antioxidants. 2017;6(4):100.
Kiamehr M, Klettner A, Richert E, Koskela A, Koistinen A, Skottman H, et al. Compromised barrier function in human induced pluripotent stem-cell-derived retinal pigment epithelial cells from type 2 diabetic patients. Int J Mol Sci. 2019;20(15):3773.
Lee D, Hong HS. Substance P alleviates retinal pigment epithelium dysfunction caused by high glucose-induced stress. Life. 2023;13(5):1070.
Sikder MM, Li X, Akumwami S, Labony SA. Reactive oxygen species: role in pathophysiology, and mechanism of endogenous and dietary antioxidants during oxidative stress. Chonnam Med J. 2025;61(1):32.
Fernandez-Robredo P, González-Zamora J, Recalde S, Bilbao-Malavé V, Bezunartea J, Hernandez M, et al. Vitamin D protects against oxidative stress and inflammation in human retinal cells. Antioxidants. 2020;9(9):838.
Markitantova Y, Simirskii V. Retinal pigment epithelium under oxidative stress: Chaperoning autophagy and beyond. Int J Mol Sci. 2023;24(14):11520.
Mustafa M, Ahmad R, Tantry IQ, Ahmad W, Siddiqui S, Alam M, et al. Apoptosis: A comprehensive overview of signaling pathways, morphological changes, and physiological significance and therapeutic implications. Cells. 2024;13(22):1838.
Gupta S, Kass GE, Szegezdi E, Joseph B. The mitochondrial death pathway: A promising therapeutic target in diseases. J Cell Mol Med. 2009;13(6):1004-33.
Dammak A, Huete-Toral F, Carpena-Torres C, Martin-Gil A, Pastrana C, Carracedo G. From oxidative stress to inflammation in the posterior ocular diseases: Diagnosis and treatment. Pharmaceutics. 2021;13(9):1376.
Liu Y, Shi Y, Han R, Liu C, Qin X, Li P, et al. Signaling pathways of oxidative stress response: The potential therapeutic targets in gastric cancer. Front Immunol. 2023;14:1139589.
You L, Peng H, Liu J, Cai M, Wu H, Zhang Z, et al. Catalpol protects ARPE-19 cells against oxidative stress via activation of the Keap1/Nrf2/ARE pathway. Cells. 2021;10(10):2635.
Prasansuklab A, Brimson JM, Tencomnao T. Potential Thai medicinal plants for neurodegenerative diseases: A review focusing on the anti-glutamate toxicity effect. J Tradit Complement Med. 2020;10(3):301-8.
Prasanth MI, Sivamaruthi BS, Sukprasansap M, Chuchawankul S, Tencomnao T, Chaiyasut C. Functional properties and bioactivities of Cleistocalyx nervosum var. paniala berry plant: A review. Food Sci Technol. 2020;40(4):779-86.
Brimson JM, Prasanth MI, Isidoro C, Sukprasansap M, Tencomnao T. Cleistocalyx nervosum var. paniala seed extracts exhibit sigma-1 antagonist sensitive neuroprotective effects in PC12 cells and protects C. elegans from stress via the SKN-1/NRF-2 pathway. Nutr Healthy Aging. 2021;6(2):131-46.
Promkum C, Mudor H, Sridonpai P, Muangnoi C, Tuntipopipat S, Sukprasansap M. Ripe Cleistocalyx nervosum var. paniala berry fruit extract prevents hydrogen peroxide-induced DNA damage in human monocyte U937 cells. Thai Bull Pharm Sci. 2025;20(2):243-58.
Panritdum P, Muangnoi C, Tuntipopipat S, Charoenkiatkul S, Sukprasansap M. Cleistocalyx nervosum var. paniala berry extract and cyanidin-3-glucoside inhibit hepatotoxicity and apoptosis. Food Sci Nutr. 2024;12(4):2947-62.
Mattioli R, Francioso A, Mosca L, Silva P. Anthocyanins: A comprehensive review of their chemical properties and health effects on cardiovascular and neurodegenerative diseases. Molecules. 2020;25(17):3809.
Gonçalves AC, Nunes AR, Falcão A, Alves G, Silva LR. Dietary effects of anthocyanins in human health: A comprehensive review. Pharmaceuticals. 2021;14(7):690.
Nomi Y, Iwasaki-Kurashige K, Matsumoto H. Therapeutic effects of anthocyanins for vision and eye health. Molecules. 2019;24(18):3311.
Duan H, Wang D, Zheng Y, Zhou Y, Yan W. The powerful antioxidant effects of plant fruits, flowers, and leaves help to improve retinal damage and support the relief of visual fatigue. Heliyon. 2024;10(14):e34153.
Maiuolo J, Bulotta RM, Oppedisano F, Bosco F, Scarano F, Nucera S, et al. Potential properties of natural nutraceuticals and antioxidants in age-related eye disorders. Life. 2023;13(1):77.
Gulcin İ, Alwasel SH. DPPH radical scavenging assay. Processes. 2023;11(8):2248.
Munsuk T, Sukboon P, Phumsuay R, Tuntipopipat S, Muangnoi C. Protective effects of extraction and bioaccessible fraction of gac fruit against H2O2-induced oxidative damage in human retinal pigment epithelial (ARPE-19) cells. Thai J Toxicol. 2024;39(1):22-36.
Asma U, Bertotti ML, Zamai S, Arnold M, Amorati R, Scampicchio M. A kinetic approach to oxygen radical absorbance capacity (ORAC): Restoring order to the antioxidant activity of hydroxycinnamic acids and fruit juices. Antioxidants. 2024;13(2):222.
Zhong Y, Shahidi F. Methods for the assessment of antioxidant activity in foods. In: Shahidi F, editor. Handbook of antioxidants for food preservation. Cambridge: Woodhead Publishing; 2015. p. 287-333.
Payne AC, Mazzer A, Clarkson GJ, Taylor G. Antioxidant assays - consistent findings from FRAP and ORAC reveal a negative impact of organic cultivation on antioxidant potential in spinach but not watercress or rocket leaves. Food Sci Nutr. 2013;1(6):439-44.
Tecce N, Cennamo G, Rinaldi M, Costagliola C, Colao A. Exploring the impact of glycemic control on diabetic retinopathy: Emerging models and prognostic implications. J Clin Med. 2024;13(3):831.
Wang L, Zhou X, Yin Y, Mai Y, Wang D, Zhang X. Hyperglycemia induces neutrophil extracellular traps formation through an NADPH oxidase-dependent pathway in diabetic retinopathy. Front Immunol. 2019;9:3076.
Xia T, Rizzolo LJ. Effects of diabetic retinopathy on the barrier functions of the retinal pigment epithelium. Vision Res. 2017;139:72-81.
Shinde PL, John S, Mishra R. Understanding the combined effects of high glucose induced hyper-osmotic stress and oxygen tension in the progression of tumourigenesis: From mechanism to anti-cancer therapeutics. Cells. 2023;12(6):825.
Stitt AW, Curtis TM, Chen M, Medina RJ, McKay GJ, Jenkins A, et al. The progress in understanding and treatment of diabetic retinopathy. Prog Retin Eye Res. 2016;51:156-86.
Wang S, Li W, Chen M, Cao Y, Lu W, Li X. The retinal pigment epithelium: Functions and roles in ocular diseases. Fundam Res. 2024;4(6):1710-8.
Carvalho F, Lahlou RA, Silva LR. Exploring bioactive compounds from fruit and vegetable by-products with potential for food and nutraceutical applications. Foods. 2025;14(22):3884.
Du B, Xu B. Natural bioactive compounds exerting health-promoting effects by ameliorating oxidative stress. Antioxidants (Basel). 2025:14(1):85.
Harju N. Regulation of oxidative stress and inflammatory responses in human retinal pigment epithelial cells [dissertation]. Kuopio: University of Eastern Finland; 2022.
Peng W, Wu Y, Peng Z, Qi W, Liu T, Yang B, et al. Cyanidin-3-glucoside improves the barrier function of retinal pigment epithelium cells by attenuating endoplasmic reticulum stress-induced apoptosis. Food Res Int. 2022;157:111313.
Tan J, Li Y, Hou DX, Wu S. The effects and mechanisms of cyanidin-3-glucoside and its phenolic metabolites in maintaining intestinal integrity. Antioxidants. 2019;8(10):479.
Frountzas M, Karanikki E, Toutouza O, Sotirakis D, Schizas D, Theofilis P, et al. Exploring the impact of cyanidin-3-glucoside on inflammatory bowel diseases: investigating new mechanisms for emerging interventions. Int J Mol Sci. 2023;24(11):9399.
Kim J, Lee YJ, Won JY. Molecular mechanisms of retinal pigment epithelium dysfunction in age-related macular degeneration. Int J Mol Sci. 2021;22(22):12298.
Gao J, Cui JZ, To E, Cao S, Matsubara JA. Evidence for the activation of pyroptotic and apoptotic pathways in RPE cells associated with NLRP3 inflammasome in the rodent eye. J Neuroinflammation. 2018;15(1):15.
Jomova K, Alomar SY, Alwasel SH, Nepovimova E, Kuca K, Valko M. Several lines of antioxidant defense against oxidative stress: Antioxidant enzymes, nanomaterials with multiple enzyme-mimicking activities, and low-molecular-weight antioxidants. Arch Toxicol. 2024;98(5):1323-67.
Xing Y, Liang S, Zhang L, Ni H, Zhang X, Wang J, et al. Combination of Lactobacillus fermentum NS9 and aronia anthocyanidin extract alleviates sodium iodate-induced retina degeneration. Sci Rep. 2023;13(1):8380.
Chaiwangyen W, Khantamat O, Pintha K, Kangwan N, Onsa-Ard A, Nuntaboon P, et al. Cleistocalyx nervosum var. paniala mitigates oxidative stress and inflammation induced by PM10 soluble extract in trophoblast cells via miR-146a-5p. Sci Rep. 2024;14(1):24265.
Nantacharoen W, Baek SJ, Plaingam W, Charoenkiatkul S, Tencomnao T, Sukprasansap M. Cleistocalyx nervosum var. paniala berry promotes antioxidant response and suppresses glutamate-induced cell death via SIRT1/Nrf2 survival pathway in hippocampal HT22 neuronal cells. Molecules. 2022;27(18):5813.
Sukprasansap M, Chanvorachote P, Tencomnao T. Cleistocalyx nervosum var. paniala berry fruit protects neurotoxicity against endoplasmic reticulum stress-induced apoptosis. Food Chem Toxicol. 2017;103:279-88.
Thuschana W, Thumvijit T, Chansakaow S, Ruamrungsri S, Wongpoomchai R. Chemical constituents and antioxidant activities of Cleistocalyx nervosum fruits in in vitro and in vivo models. Thai J Toxicol. 2012;27(2):194-207.
Kowalski R, Gustafson E, Carroll M, de Gonzalez M. Enhancement of biological properties of blackcurrants by lactic acid fermentation and incorporation into yogurt: A review. Antioxidants. 2020;9(12):1194.
Castilho ÁF, Aveleira CA, Leal EC, Simões NF, Fernandes CR, Meirinhos RI, et al. Heme oxygenase-1 protects retinal endothelial cells against high glucose- and oxidative/ nitrosative stress-induced toxicity. PLoS One. 2012;7(8): e42428.
Mecchia A, Palumbo C, De Luca A, Sbardella D, Boccaccini A, Rossi L, et al. High glucose induces an early and transient cytoprotective autophagy in retinal Müller cells. Endocrine. 2022;77(2):221-30.
Janani R, Anitha RE, Divya P, Chonche M, Baskaran V. Astaxanthin ameliorates hyperglycemia induced inflammation via PI3K/Akt–NF–κB signaling in ARPE-19 cells and diabetic rat retina. Eur J Pharmacol. 2022;926:174979.
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