The SH-SY5Y Human Cell Line: Hawthorne Berry (Crataegus spp.) Protects against 6-OHDA Induced Neurotoxicity In Vitro Model of Parkinson's Disease

Abstract

Aim: We purposed to study the neuroprotective effects of Hawthorn berry (crataegus spp.) extract, which is familiar to have antioxidant and anti-inflammatory features, opposite the neurotoxicity led to by 6-OHDA in SH-SY5Y cells. Method: SH-SY5Y cells were treated with Hawthorn berry (25-50-75 and 100 μg/mL) for two hours ago 6-OHDA administration. Cells were exposed to 200 µM 6-OHDA for 24 hours to mimic the in vitro Parkinson's disease model. After one day, cell viability was measured by lactate dehydrogenase and 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide analysis. Oxidative stress was evaluated with tumor necrosis factor-α, interleukin-1β, superoxide dismutase, catalase, glutathione, glutathione peroxidase, myeloperoxidase, and malondialdehyde assays. Results: It was found that the viability rate of Hawthorn berry increased depending on the concentration and the cell viability was 94% at the highest concentration (p<0.001). Also, 6-OHDA raised lactate dehydrogenase leakage in SH-SY5Y cells (p<0.001). While 6-OHDA exacerbated oxidative stress by enhancing tumor necrosis factor-α, interleukin-1β, myeloperoxidase, and malondialdehyde (p<0.001), pretreatment with Hawthorn berry alleviated these toxic effects of 6-OHDA through antioxidant capacity by increasing glutathione peroxidase, superoxide dismutase, catalase and glutathione (p<0.05), (p<0.001). In line with all findings, Hawthorn berry attenuated neuronal cell demise in a dose-dependent manner. Conclusion: Considering its neuroprotective role as well as its effects on oxidative stress, Hawthorn berry could be a potential natural bio-medicine to prevent the development of Parkinson's disease.

Keywords

• Antioxidant
• Hawthorn berry
• 6-OHDA
• Parkinson’s disease
• SH-SY5Y cell line

SH-SY5Y İnsan Hücre Hattı: Hawthorne Berry (Crataegus spp.) Parkinson Hastalığının İn Vitro Modelinde Oluşturulan 6-OHDA Kaynaklı Nörotoksisiteye Karşı Korur

Abstract

Amaç: Çalışmada antioksidan ve antiinflamatuar özelliklere sahip olduğu bilinen Hawthorn berry (crataegus spp.) ekstraktın, SH-SY5Y hücrelerinde 6-OHDA ile meydana gelen nörotoksisiteye karşı nöroprotektif etkilerini araştırmayı amaçlanmıştır. Yöntem: SH-SY5Y hücreleri, 6-OHDA uygulamasından önce iki saat boyunca Hawthorn berry (25, 50, 75 ve 100 μg/mL) ile muamele edildi. Hücreler, in vitro Parkinson hastalığı modelini taklit etmek için 24 saat boyunca 200 uM 6-OHDA’ya maruz bırakıldı. Bir gün sonra, 3-(4,5 Dimetiltiazol-2-il)-2,5-difeniltetrazolyum bromür ve laktat dehidrogenaz tahlilleri ile hücre canlılığı belirlendi. Oksidatif stres tümör nekroz faktör-α, interlökin-1β, süperoksit dismutaz, katalaz, glutatyon, glutatyon peroksidaz, miyeloperoksidaz ve malondialdehit analizleri ile değerlendirildi. Bulgular: Canlılık oranında Hawthorn berry’nin konsantrasyona bağlı olarak bir artış gösterdiği ve en yüksek konsantrasyonda hücre canlılığı % 94 oranında bulundu (p<0,001). Ayrıca, 6-OHDA SH-SY5Y hücrelerinde laktat dehidrogenaz sızıntısını arttırdı (p<0,001). 6-OHDA tümör nekroz faktör-α, interlökin-1β, miyeloperoksidaz ve malondialdehit artırarak oksidatif stresi şiddetlendirirken (p<0,001), Hawthorn berry ile ön tedavi süperoksit dismutaz, katalaz, glutatyon ve glutatyon peroksidazı artırarak antioksidan kapasite yoluyla 6-OHDA'nın bu toksik etkilerini hafifletti (p<0,05), (p<0,001). Tüm bulgular doğrultusunda Hawthorn berry nöronal hücre ölümünü hafifleterek doza bağlı bir şekilde önledi. Sonuç: Oksidatif stres üzerindeki etkilerinin yanı sıra nöroprotektif rolü göz önüne alındığında Hawthorn berry, Parkinson hastalığının gelişimini önlemek için potansiyel doğal biyo-ilaç olabilir.

Keywords

• Antioksidan
• Hawthorn berry
• 6-OHDA
• Parkinson hastalığı
• SH-SY5Y hücre hattı

References

1. 1. Lee YY, Li MH, Tai CH, Luh JJ. Corticomotor excitability changes associated with freezing of gait in people with parkinson disease. Front Hum Neurosci. 2020;14:190.
2. 2. Ascherio A, Schwarzschild MA. The epidemiology of Parkinson’s disease: Risk factors and prevention. The Lancet. Neurology. 2016;15:1257–1272.
3. 3. Jayaram S, Krishnamurthy PT. Role of microgliosis, oxidative stress and associated neuroinflammation in the pathogenesis of Parkinson’s disease: The therapeutic role of Nrf2 activators. Neurochem Int. 2021;1(145):105014.
4. 4. Bo RX, Li YY, Zhou TT, Chen NH, Yuan YH. The neuroinflammatory role of glucocerebrosidase in Parkinson’s disease. Neuropharmacology. 2022;19:108964.
5. 5. Piancone F, Saresella M, La Rosa F, et al. Inflammatory responses to monomeric and aggregated α-synuclein in peripheral blood of Parkinson disease patients. Front Neurosci. 2021;15:639646.
6. 6. Dionísio PA, Amaral JD, Rodrigues CM. Oxidative stress and regulated cell death in Parkinson’s disease. Ageing Res Rev. 2021;1(67):101263.
7. 7. Sodhi RK, Bansal Y, Singh R, et al. IDO-1 inhibition protects against neuroinflammation, oxidative stress and mitochondrial dysfunction in 6-OHDA induced murine model of Parkinson’s disease. Neurotoxicology. 2021;1(84):184–197.
8. 8. Hernandez-Baltazar D, Zavala-Flores LM, Villanueva-Olivo A. The 6-hydroxydopamine model and parkinsonian pathophysiology: Novel findings in an older model. Neurologia. 2017;32:533–539.

9. 9. Alrashidi H, Eaton S, Heales S. Biochemical characterization of proliferative and differentiated SH-SY5Y cell line as a model for Parkinson’s disease. Neurochem Int. 2021;1(145):105009.
10. 10. Huang XX, Ren Q, Song XY, et al. Seven new sesquineolignans isolated from the seeds of hawthorn and their neuroprotective activities. Fitoterapia. 2018;125:6-12.
11. 11. Li C, Wang MH. Anti-inflammatory effect of the water fraction from hawthorn fruit on LPS-stimulated RAW 264.7 cells. Nutrition Research and Practice. 2011;5(2):101–106.
12. 12. Niu ZZ, Yan MX, Zhao XL, Jin H, Gong YL. Effect of hawthorn seed extract on the gastrointestinal function of rats with diabetic gastroparesis. S Afr J Bot. 2020;130:448–455.
13. 13. Dolatkhani P, Jameie R. Antioxidant poperties and medicinal uses of some crataegus spp. (Hawthorn) including C. Meyeri and C. Pontica Curr. Nutr. Food Sci. 2015;11(2):116-123.
14. 14. Ruan WL, Shen SH, Xu Y, Ran N, Zhang H. Mechanistic insights into procyanidins as therapies for Alzheimer’s disease: A review. J Funct Foods. 2021;86-104683.
15. 15. Tan JW, Kim MK. Neuroprotective effects of bochanin A against β-amyloid-induced neurotoxicity in PC12 cells via a mitochondrial-dependent apoptosis pathway. Molecules. 2016;21:548.
16. 16. Olatunji OJ, Feng Y, Olatunji OO, Tang J, Ouyang Z, Su Z. Cordycepin protects PC12 cells against 6-hydroxydopamine induced neurotoxicity via its antioxidant properties. Biomed Pharmacother. 2016;81:7-14.
17. 17. Xu Z, Yang D, Huang X, Huang H. Astragaloside IV protects 6-hydroxydopamine induced SH-SY5Y cell model of Parkinson's disease via activating the JAK2/STAT3 pathway. Front Neurosci. 2021;15:631501.
18. 18. Angelova PR, Abramov AY. Role of mitochondrial ROS in the brain: From physiology to neurodegeneration. FEBS Lett. 2018;592:692–702.
19. 19. Redza-Dutordoir M, Averill-Bates DA. Activation of apoptosis signalling pathways by reactive oxygen species. Biochim Biophys Acta. 2016;1863:2977–2992.
20. 20. Ahiskali I, Ferah Okkay I, Mammadov R, et al. Effect of taxifolin on cisplatin-associated oxidative optic nerve damage in rats. Cutan Ocul Toxicol. 2021;40:1–6.
21. 21. Wang HL, Zhang J, Li YP, Dong L, Chen YZ. Potential use of glutathione as a treatment for Parkinson’s disease. Exp Ther Med. 2021;21:1–1.
22. 22. Ko YH, Kim SK, Kwon SH, et al. 7, 8, 4′-Trihydroxyisoflavone, a metabolized product of daidzein, attenuates 6-hydroxydopamine-induced neurotoxicity in SH-SY5Y cells. Biomol Ther. 2019;27:363.
23. 23. Antunes MS, Goes AT, Boeira SP, Prigol M, Jesse CR. Protective effect of hesperidin in a model of Parkinson’s disease induced by 6-hydroxydopamine in aged mice. Nutrition. 2014;30:1415–1422.
24. 24. Porres-Martínez M, González-Burgos E, Carretero ME, Gómez-Serranillos MP. Protective properties of Salvia lavandulifolia Vahl. essential oil against oxidative stress-induced neuronal injury. Food Chem Toxicol. 2015;80:154–162.
25. 25. Romuk E, Szczurek W, Nowak P, et al. Effects of propofol on oxidative stress parameters in selected parts of the brain in a rat model of Parkinson disease. Postepy Hig Med Dosw. 2016;70(0):1441-1450.
26. 26. Chen G, Liu J, Jiang L, et al. Galangin reduces the loss of dopaminergic neurons in an LPS-evoked model of Parkinson’s disease in rats. Int. J. Mol. Sci. 2018;19(1):12.
27. 27. Cheng J, Duan Y, Zhang F, et al. The role of lncRNA TUG1 in the Parkinson disease and its effect on microglial inflammatory response. Neuromolecular Med. 2021;23 (2):327–334.
28. 28. Zorov DB, Juhaszova M, Sollott SJ. Mitochondrial reactive oxygen species (ROS) and ROS-induced ROS release. Physiol Rev. 2014;94(3):909-950.
29. 29. Cirmi S, Maugeri A, Lombardo GE, et al. A flavonoidrich extract of mandarin juice counteracts 6-OHDAinduced oxidative stress in SH-SY5Y cells and modulates Parkinson-related genes. Antioxidants (Basel). 2021;10(4):539.