Apigenin-7-Glikozit Antioksidan Enzim Sisteminin Aktivasyonu ve Kaspaz Genlerinin Ekspresyonunun İnhibisyonu Yoluyla SH-SY5Y Hücrelerinde Hidrojen Peroksit İndüklü Oksidatif Stresi ve Nöronal Ölümü Azaltır

Yıl 2023, Cilt: 33 Sayı: 2, 162 - 168, 30.04.2023

Ebru Güçlü , İlknur Çınar Ayan

https://doi.org/10.54005/geneltip.1219084

Öz

ÖZ
Amaç: Bu çalışmada antikanser ve anti-inflamatuar etkinliği bilinen bir doğal ürün olan Apigenin-7-Glikozitin (AP7Glu), SH-SY5Y hücrelerinde hidrojen proksit (H2O2)-indüklü oksidatif strese karşı olası nöroprotektif etkisinin araştırılması amaçlandı.
Yöntem: H2O2 ve AP7Glu’nun hücre canlılığı üzerine etkisi XTT analizi ile belirlendi. Antioksidan enzim aktivitelerinin belirlenmesine yönelik olarak SOD ve katalaz analizleri gerçekleştirildi. Antioksidan savunma sistemi ile ilişkili genler ile Kaspaz genlerinin ekspresyon seviyeleri qRT-PCR analizi ile değerlendirildi.
Bulgular: SH-SY5Y hücrelerine 24 saat süresince 300 µM H2O2 uygulaması sonrasında hücre canlılığı % 63.18 olarak belirlendi ve oksidatif stresin indüklenmesi için bu doz kullanıldı. Farklı konsantrasyonlarda AP7Glu varlığında H2O2 uygulaması sadece H2O2 ile muamele edilmiş hücreler ile karşılaştırıldığında hücre canlılığını artırdı. H2O2 SOD ve katalaz enzim aktivitelerini kontrol grubuna göre azaltttı. H2O2’nin AP7Glu varlığında uygulanması ise sadece H2O2 ile muamele edilmiş hücreler ile karşılaştırıldığında bu enzimlerin aktivitelerinde artış gözlendi. Ayrıca, H2O2 antioksidan savunma sistemi ile ilişkili genlerin ekspresyon seviyelerini azaltırken Kaspaz genlerinin ekspresyon seviyelerini artırdı. Sadece H2O2 ile muamele edilmiş hücreler ile karşılaştırıldığında ise AP7Glu varlığında H2O2 uygulaması antioksidan savunma sistemi ile ilişkili genlerin ekspresyon seviyelerini arttırdı ve CASP8 ekspresyonunu azalttı.
Sonuç: AP7Glu H2O2 ile indüklenmiş oksidatif stres ve nöronal hücre ölümüne karşı nöroprotektif bir etkiye sahiptir.

Anahtar Kelimeler

Hidrojen peroksit, Oksidatif stres, Apigenin-7-Glikozit, SH-SY5Y

Apigenin-7-Glucoside Attenuates Hydrogen Peroxide-Induced Oxidative Stress and Neuronal Death in SH-SY5Y Cells Via Activation of Antioxidant Enzymes System and Inhibition of Caspases Genes Expression

Öz

Aims: In this study, it was aimed to investigate the possible neuroprotective effect of Apigenin-7-Glycoside (AP7Glu), a natural product with known anticancer and anti-inflammatory activity, against hydrogen proxide (H2O2)-induced oxidative stress in SH-SY5Y cells.
Methods: The effect of H2O2 and AP7Glu on cell viability was determined by XTT analysis. SOD and catalase analyzes were performed to determine antioxidant enzyme activities. Expression levels of antioxidant defense system related genes and caspase genes were evaluated by qRT-PCR analysis.
Results: After treatment of 300 µM H2O2 to SH-SY5Y cells for 24 hours, cell viability was determined as 63.18% and this dose was used to induce oxidative stress. H2O2 treatment in the presence of different concentrations of AP7Glu increased cell viability compared to H2O2-treated cells. And, H2O2 decreased SOD and catalase enzyme activities compared to control group. With the treatment of H2O2 in the presence of AP7Glu, an increase was observed in these enzymes activities compared to H2O2-treated cells. In addition, H2O2 decreased the expression levels of genes associated with antioxidant defense system and increased expression levels of Caspase genes. And, H2O2 treatment in the presence of AP7Glu increased the expression levels of genes associated with antioxidant defense system and decreased CASP8 expression compared to H2O2-treated cells.
Conclusion: AP7Glu has a neuroprotective effect against H2O2-induced oxidative stress and neuronal cell death.

Anahtar Kelimeler

Apigenin-7-Glycoside, Hydrogen peroxide, Oxidative stress, SH-SY5Y, Apigenin-7-Glikozit, Hidrojen peroksit, Oksidatif stres

Kaynakça

• Jenner P. Oxidative stress in Parkinson's disease. Ann Neurol 2003;53 Suppl 3:S26-36
• Smith MA, Rottkamp CA, Nunomura A, Raina AK, Perry G. Oxidative stress in Alzheimer's disease. Biochim Biophys Acta 2000;1502(1):139-144
• Overmyer K, Brosché M, Kangasjärvi J. Reactive oxygen species and hormonal control of cell death. Trends Plant Sci 2003;8(7):335-342
• Crane FL, Low H. Reactive oxygen species generation at the plasma membrane for antibody control. Autoimmun Rev 2008;7(7):518-522
• Bhandary B, Marahatta A, Kim HR, Chae HJ. Involvement of oxidative stress in endoplasmic reticulum stress and its associated diseases. Int J Mol Sci 2012;14(1):434-456
• Pizzino G, Irrera N, Cucinotta M, Pallio G, Mannino F, Arcoraci V, Squadrito F, Altavilla D, Bitto A. Oxidative Stress: Harms and Benefits for Human Health. Oxid Med Cell Longev 2017;2017:8416763
• Yavarpour-Bali H, Ghasemi-Kasman M, Pirzadeh M. Curcumin-loaded nanoparticles: a novel therapeutic strategy in the treatment of central nervous system disorders. Int J Nanomedicine 2019;14:4449-4460
• Collin F. Chemical Basis of Reactive Oxygen Species Reactivity and Involvement in Neurodegenerative Diseases. Int J Mol Sci 2019;20(10):2407
• Annunziato L, Amoroso S, Pannaccione A, Cataldi M, Pignataro G, D'Alessio A, Sirabella R, Secondo A, Sibaud L, Di Renzo GF. Apoptosis induced in neuronal cells by oxidative stress: the role played by caspases and intracellular calcium ions. Toxicol Lett 2003;139(2-3):125-133
• Singh A, Kukreti R, Saso L, Kukreti S. Oxidative Stress: A Key Modulator in Neurodegenerative Diseases. Molecules 2019;24(8):1583
• Jantas D, Chwastek J, Grygier B, Lasoń W. Neuroprotective Effects of Necrostatin-1 Against Oxidative Stress-Induced Cell Damage: an Involvement of Cathepsin D Inhibition. Neurotox Res 2020;37(3):525-542
• Ruankham W, Suwanjang W, Wongchitrat P, Prachayasittikul V, Prachayasittikul S, Phopin K. Sesamin and sesamol attenuate H2O2-induced oxidative stress on human neuronal cells via the SIRT1-SIRT3-FOXO3a signaling pathway. Nutr Neurosci 2021;24(2):90-101
• Asaduzzaman Khan M, Tania M, Zhang DZ, Chen HC. Antioxidant enzymes and cancer. Chin J Cancer Res 2010;22:87-92
• Zhang H, Rong T. Dietary polyphenols, oxidative stress and antioxidant and anti-inflammatory effects. Current Opinion in Food Science 2016, 8: 33-42
• Uddin MS, Mamun AA, Rahman MM, Jeandet P, Alexiou A, Behl T, Sarwar MS, Sobarzo-Sánchez E, Ashraf GM, Sayed AA, Albadrani GM, Peluso I, Abdel-Daim MM. Natural Products for Neurodegeneration: Regulating Neurotrophic Signals. Oxid Med Cell Longev 2021;2021:8820406
• Perrott KM, Wiley CD, Desprez PY, Campisi J. Apigenin suppresses the senescence-associated secretory phenotype and paracrine effects on breast cancer cells. Geroscience 2017;39(2):161-173.
• Sánchez-Marzo N, Pérez-Sánchez A, Ruiz-Torres V, Martínez-Tébar A, Castillo J, Herranz-López M, Barrajón-Catalán E. Antioxidant and Photoprotective Activity of Apigenin and its Potassium Salt Derivative in Human Keratinocytes and Absorption in Caco-2 Cell Monolayers. Int J Mol Sci 2019;20(9):2148
• Nayaka HB, Londonkar RL, Umesh MK, Tukappa A. Antibacterial Attributes of Apigenin, Isolated from Portulaca oleracea L. Int J Bacteriol 2014;2014:175851
• Sadraei H, Asghari G, Khanabadi M, Minaiyan M. Anti-inflammatory effect of apigenin and hydroalcoholic extract of Dracocephalum kotschyi on acetic acid-induced colitis in rats. Res Pharm Sci 2017;12(4):322-329
• Li KC, Ho YL, Hsieh WT, Huang SS, Chang YS, Huang GJ. Apigenin-7-glycoside prevents LPS-induced acute lung injury via the downregulation of oxidative enzyme expression and protein activation through the inhibition of MAPK phosphorylation. Int J Mol Sci 2015;16(1):1736-1754
• Güçlü E, Çınar Ayan İ, Dursun HG, Vural H. Tomentosin induces apoptosis in pancreatic cancer cells through increasing reactive oxygen species and decreasing mitochondrial membrane potential. Toxicol In Vitro;84:105458.
• Wang J, Zhao YM, Zhang B, Guo CY. Protective effect of total phenolic compounds from Inula helenium on hydrogen peroxide-induced oxidative stress in SH-SY5Y cells. Indian journal of pharmaceutical sciences 2015;77(2):163.
• Tian W, Heo S, Kim DW, Kim IS, Ahn D, Tae HJ, Kim MK, Park BY. Ethanol Extract of Maclura tricuspidata Fruit Protects SH-SY5Y Neuroblastoma Cells against H2O2-Induced Oxidative Damage via Inhibiting MAPK and NF-κB Signaling. Int J Mol Sci 2021;22(13):6946
• Smiljkovic M, Stanisavljevic D, Stojkovic D, Petrovic I, Marjanovic Vicentic J, Popovic J, Golic Grdadolnik S, Markovic D, Sankovic-Babice S, Glamoclija J, Stevanovic M, Sokovic M. Apigenin-7-O-glucoside versus apigenin: Insight into the modes of anticandidal and cytotoxic actions. EXCLI J 2017;16:795-807
• Liu MM, Ma RH, Ni ZJ, Thakur K, Cespedes-Acuña CL, Jiang L, Wei ZJ. Apigenin 7-O-glucoside promotes cell apoptosis through the PTEN/PI3K/AKT pathway and inhibits cell migration in cervical cancer HeLa cells. Food Chem Toxicol 2020;146:111843
• Tian W, Zhao J, Lee JH, Akanda MR, Cho JH, Kim SK, Choi YJ, Park BY. Neuroprotective Effects of Cornus officinalis on Stress-Induced Hippocampal Deficits in Rats and H2O2-Induced Neurotoxicity in SH-SY5Y Neuroblastoma Cells. Antioxidants (Basel) 2019;9(1):27
• Younus H. Therapeutic potentials of superoxide dismutase. Int J Health Sci (Qassim) 2018;12(3):88-93
• Islam MN, Rauf A, Fahad FI, Emran TB, Mitra S, Olatunde A, Shariati MA, Rebezov M, Rengasamy KRR, Mubarak MS. Superoxide dismutase: an updated review on its health benefits and industrial applications. Crit Rev Food Sci Nutr 2022;62(26):7282-7300
• Ighodaro OM, Akinloye OA. First-line defense antioxidants-superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX): Their fundamental role in the entire antioxidant defense grid. Alexandria journal of medicine 2018;54(4): 287-293.
• Feng C, Luo T, Zhang S, Liu K, Zhang Y, Luo Y, Ge P. Lycopene protects human SH‑SY5Y neuroblastoma cells against hydrogen peroxide‑induced death via inhibition of oxidative stress and mitochondria‑associated apoptotic pathways. Mol Med Rep 2016;13(5):4205-4214
• Gao J, Xu Y, Zhang J, Shi J, Gong Q. Lithocarpus polystachyus Rehd. leaves aqueous extract protects against hydrogen peroxide‑induced SH-SY5Y cell injury through activation of the Sirt3 signaling pathway. Int J Mol Med 2018;42(6):3485-3494
• Ureña-Vacas I, González-Burgos E, Divakar PK, Gómez-Serranillos MP. Lichen Extracts from Cetrarioid Clade Provide Neuroprotection against Hydrogen Peroxide-Induced Oxidative Stress. Molecules 2022;27(19):6520
• Nojima Y, Ito K, Ono H, Nakazato T, Bono H, Yokoyama T, Sato R, Suetsugu Y, Nakamura Y, Yamamoto K, Satoh J, Tabunoki H, Fugo H. Superoxide dismutases, SOD1 and SOD2, play a distinct role in the fat body during pupation in silkworm Bombyx mori. PLoS One 2015;10(2):e0116007
• Pansarasa O, Bordoni M, Diamanti L, Sproviero D, Gagliardi S, Cereda C. SOD1 in Amyotrophic Lateral Sclerosis: “Ambivalent” Behavior Connected to the Disease. International Journal of Molecular Sciences 2018; 19(5):1345
• Alateyah N, Gupta I, Rusyniak RS, Ouhtit A. SOD2, a Potential Transcriptional Target Underpinning CD44-Promoted Breast Cancer Progression. Molecules 2022;27(3):811
• Erdman VV, Nasibullin TR, Tuktarova IA, Timasheva YR, Danilko KV, Viktorova, TV, Mustafina OE. The Study of Association of Polymorphic Markers of the SOD1, SOD2, and SOD3 Genes with Longevity. Russian Journal of Genetics 2020;56(12): 1504-1511
• Sah SK, Agrahari G, Kim TY. Insights into superoxide dismutase 3 in regulating biological and functional properties of mesenchymal stem cells. Cell Biosci 2020;10:22
• Nandi A, Yan LJ, Jana CK, Das N. Role of Catalase in Oxidative Stress- and Age-Associated Degenerative Diseases. Oxid Med Cell Longev 2019;2019:9613090
• Méndez-Armenta M, Nava-Ruíz C, Juárez-Rebollar D, Rodríguez-Martínez E, Gómez PY. Oxidative stress associated with neuronal apoptosis in experimental models of epilepsy. Oxid Med Cell Longev 2014;2014:293689
• Li J, Yuan J. Caspases in apoptosis and beyond. Oncogene 2008;27(48):6194-6206
• Fiandalo MV, Kyprianou N. Caspase control: protagonists of cancer cell apoptosis. Exp Oncol 2012;34(3):165-175