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Neuroprotective effects of cerebroprotein hydrolysate and its combination with antioxidants against oxidative stress-induced HT22 cell death

  • Eun‑Ju Yang (College of Pharmacy, Chung-Ang University) ;
  • Jae Cheon Kim (Department of Global Innovative Drugs, The Graduate School of Chung-Ang University) ;
  • Dong Hee Na (College of Pharmacy, Chung-Ang University)
  • Received : 2024.03.07
  • Accepted : 2024.05.15
  • Published : 2024.10.15

Abstract

This study aimed to investigate the neuroprotective effects of cerebroprotein hydrolysate (CPH) against oxidative stress-induced HT22 cell death. Additionally, the effect of antioxidants such as quercetin (QC) and N-acetyl-L-cysteine (NAC) on the neuroprotective activity of CPH was evaluated. The mouse-derived hippocampal neuronal cell line HT22 was pretreated with CPH or a mixture of CPH and QC or NAC. HT22 cell death was induced by either 10 mM glutamate, 2.5 μM amyloid-β (Aβ)25-35, and 300 μM cobalt chloride (CoCl2). As results, CPH effectively alleviated HT22 cell death induced by glutamate, Aβ25-35, and CoCl2. In addition, CPH combination with QC augmented cell viability in both glutamate- and Aβ25-35-stressed conditions but had no synergic effect on the CoCl2-stressed condition. The synergic effect of CPH and NAC combination was observed under all cell death conditions. The neuroprotective actions of CPH and its combinations with QC or NAC against various oxidative stress-induced HT22 cell deaths were demonstrated, providing a promising strategy for developing CPH preparations for the prevention and/or treatment of neurodegenerative diseases such as Alzheimer's disease.

Keywords

Acknowledgement

This study was supported by Daewoong Bio Inc.

References

  1. Fiani B, Covarrubias C, Wong A, Doan T, Reardon T, Nikolaidis D, Sarno E (2021) Cerebrolysin for stroke, neurodegeneration, and traumatic brain injury: review of the literature and outcomes. Neurol Sci 42:1345-1353. https://doi.org/10.1007/s10072-021-05089-2 
  2. Wu X, Liu Y, Zhu L, Wang Y, Ren Y, Cheng B, Ren L, Ge K, Li H (2021) Cerebroprotein hydrolysate-I inhibits hippocampal neuronal apoptosis by activating PI3K/Akt signaling pathway in vascular dementia mice. Neuropsychiatr Dis Treat 17:2359-2368. https://doi.org/10.2147/NDT.S311760 
  3. Liu Z, Wang W, Huang T, Wang C, Huang Y, Tang Y, Huang J (2019) CH(II), a cerebroprotein hydrolysate, exhibits potential neuro-protective effect on Alzheimer's disease. PLoS One 14:e0222757. https://doi.org/10.1371/journal.pone.0222757 
  4. Xing S, Zhang J, Dang C, Liu G, Zhang Y, Li J, Fan Y, Pei Z, Zeng J (2014) Cerebrolysin reduces amyloid-beta deposits, apoptosis and autophagy in the thalamus and improves functional recovery after cortical infarction. J Neurol Sci 337:104-111. https://doi.org/10.1016/j.jns.2013.11.028 
  5. Ma W, Wang S, Liu X, Tang F, Zhao P, Cheng K, Zheng Q, Zhuo Y, Zhao X, Li X, Feng W (2019) Protective effect of troxerutin and cerebroprotein hydrolysate injection on cerebral ischemia through inhibition of oxidative stress and promotion of angiogenesis in rats. Mol Med Rep 19:3148-3158. https://doi.org/10.3892/mmr.2019.9960 
  6. Yao PJ, Manor U, Petralia RS, Brose RD, Wu RT, Ott C, Wang YX, Charnof A, Lippincott-Schwartz J, Mattson MP (2017) Sonic hedgehog pathway activation increases mitochondrial abundance and activity in hippocampal neurons. Mol Biol Cell 28:387-395. https://doi.org/10.1091/mbc.E16-07-0553 
  7. Chandel NS, McClintock DS, Feliciano CE, Wood TM, Melendez JA, Rodriguez AM, Schumacker PT (2000) Reactive oxygen species generated at mitochondrial complex III stabilize hypoxia-inducible factor-1alpha during hypoxia: a mechanism of O2 sensing. J Biol Chem 275:25130-25138. https://doi.org/10.1074/jbc.M001914200 
  8. Nguyen TD, Dang LN, Jang JH, Park S (2023) Recent advances in Alzheimer's disease pathogenesis and therapeutics from an immune perspective. J Pharm Investig 53:667-684. https://doi.org/10.1007/s40005-023-00631-0 
  9. Esposito Z, Belli L, Toniolo S, Sancesario G, Bianconi C, Martorana A (2013) Amyloid beta, glutamate, excitotoxicity in Alzheimer's disease: are we on the right track? CNS Neurosci Ther 19:549-555. https://doi.org/10.1111/cns.12095 
  10. Neves D, Salazar IL, Almeida RD, Silva RM (2023) Molecular mechanisms of ischemia and glutamate excitotoxicity. Life Sci 328:121814. https://doi.org/10.1016/j.lfs.2023.121814 
  11. Zhang Y, Bhavnani BR (2006) Glutamate-induced apoptosis in neuronal cells is mediated via caspase-dependent and independent mechanisms involving calpain and caspase-3 proteases as well as apoptosis inducing factor (AIF) and this process is inhibited by equine estrogens. BMC Neurosci 7:49. https://doi.org/10.1186/1471-2202-7-49 
  12. Kim HB, Yoo JY, Yoo SY, Lee JH, Chang W, Kim HS, Baik TK, Woo RS (2020) Neuregulin-1 inhibits CoCl2-induced upregulation of excitatory amino acid carrier 1 expression and oxidative stress in SH-SY5Y cells and the hippocampus of mice. Mol Brain 13:153. https://doi.org/10.1186/s13041-020-00686-2 
  13. Munoz-Sanchez J, Chanez-Cardenas ME (2019) The use of cobalt chloride as a chemical hypoxia model. J Appl Tox 39:556-570. https://doi.org/10.1002/jat.3749 
  14. Peng C, Rao W, Zhang L, Wang K, Hui H, Wang L, Su N, Luo P, Hao YL, Tu Y, Zhang S, Fei Z (2015) Mitofusin 2 ameliorates hypoxia-induced apoptosis via mitochondrial function and signaling pathways. Int J Biochem Cell Biol 69:29-40. https://doi.org/10.1016/j.biocel.2015.09.011 
  15. Guan T, Zheng Y, Jin S, Wang S, Hu M, Liu X, Huang S, Liu Y (2022) Troxerutin alleviates kidney injury in rats via PI3K/AKT pathway by enhancing MAP4 expression. Food Nutr Res 66:8469. https://doi.org/10.29219/fnr.v66.8469 
  16. Zhao H, Liu Y, Zeng J, Li D, Zhang W, Huang Y (2018) Troxerutin and cerebroprotein hydrolysate injection protects neurovascular units from oxygen-glucose deprivation and reoxygenation-induced injury in vitro. Evid Based Complement Alternat Med 2018:9859672. https://doi.org/10.1155/2018/9859672 
  17. Zhao H, Liu Y, Zeng J, Li D, Huang Y (2018) Troxerutin cerebroprotein hydrolysate injection ameliorates neurovascular injury induced by traumatic brain injury-via endothelial nitric oxide synthase pathway regulation. Int J Neurosci 128:1118-1127. https://doi.org/10.1080/00207454.2018.1486828 
  18. Zhao H, Wang R, Zhang Y, Liu Y, Huang Y (2021) Neuroprotective effects of troxerutin and cerebroprotein hydrolysate injection on the neurovascular unit in a rat model of Middle cerebral artery occlusion. Int J Neurosci 131:264-278. https://doi.org/10.1080/00207454.2020.1738431 
  19. Liang KS, Yin CB, Peng LJ, Zhang JL, Guo X, Liang SY, Zhou XY, Yuan DC, Li GL, Hu FY (2017) Effect of troxerutin and cerebroprotein hydrolysate injection for the treatment of acute cerebral infarction: a multi-center randomized, single-blind and placebo-controlled study. Int J Clin Exp Med 10:10959-10964 
  20. Yang EJ, Kim GS, Kim JA, Song KS (2013) Protective effects of onion-derived quercetin on glutamate-mediated hippocampal neuronal cell death. Pharmacogn Mag 9:302-308. https://doi.org/10.4103/0973-1296.117824 
  21. Kim HT, Yoo M, Yang EJ, Song KS, Park EJ, Na DH (2023) The importance of pH for the formation of stable and active quercetin-polyamidoamine dendrimer complex. Bull Korean Chem Soc 44:363-369. https://doi.org/10.1002/bkcs.12669 
  22. Liu J, Wei AH, Liu TT, Ji XH, Zhang Y, Yan F, Chen MX, Hu JB, Zhou SY, Shi JS (2024) Icariin ameliorates glycolytic dysfunction in Alzheimer's disease models by activating the Wnt/β-catenin signaling pathway. FEBS J 291:2221-2241. https://doi.org/10.1111/febs.17099 
  23. Guimaraes CC, Oliveira DD, Valdevite M, Saltoratto AL, Pereira SI, Franca Sde C, Pereira AM, Pereira PS (2015) The glycosylated flavonoids vitexin, isovitexin, and quercetrin isolated from Serjania erecta Radlk (Sapindaceae) leaves protect PC12 cells against amyloid-beta25-35 peptide-induced toxicity. Food Chem Toxicol 86:88-94. https://doi.org/10.1016/j.fct.2015.09.002 
  24. Lan AP, Xiao LC, Yang ZL, Yang CT, Wang XY, Chen PX, Gu MF, Feng JQ (2012) Interaction between ROS and p38MAPK contributes to chemical hypoxia-induced injuries in PC12 cells. Mol Med Rep 5:250-255. https://doi.org/10.3892/mmr.2011.623 
  25. Park JS, Park JH, Kim KY (2019) Neuroprotective effects of myristargenol A against glutamate-induced apoptotic HT22 cell death. RSC Adv 9:31247-31254. https://doi.org/10.1039/c9ra05408a 
  26. Bianchi MG, Bardelli D, Chiu M, Bussolati O (2014) Changes in the expression of the glutamate transporter EAAT3/EAAC1 in health and disease. Cell Mol Life Sci 71:2001-2015. https://doi.org/10.1007/s00018-013-1484-0 
  27. Moussawi K, Riegel A, Nair S, Kalivas PW (2011) Extracellular glutamate: functional compartments operate in different concentration ranges. Front Syst Neurosci 5:94. https://doi.org/10.3389/fnsys.2011.00094 
  28. Al-Nasser MN, Mellor IR, Carter WG (2022) Is L-glutamate toxic to neurons and thereby contributes to nuronal loss and neurodegeneration? A systematic review. Brain Sci 12:577. https://doi.org/10.3390/brainsci12050577 
  29. Talevi A (2015) Multi-target pharmacology: possibilities and limitations of the "skeleton key approach" from a medicinal chemist perspective. Front Pharmacol 6:205. https://doi.org/10.3389/fphar.2015.00205 
  30. Kang DW, Cho SJ, Choi GW, Cho HY (2022) Strategies for developing Alzheimer's disease treatments: application of population pharmacokinetic and pharmacodynamic models. J Pharm Investig 52:519-538. https://doi.org/10.1007/s40005-022-00579-7 
  31. Makhoba XH, Viegas C Jr, Mosa RA, Viegas FPD, Pooe OJ (2020) Potential impact of the multi-target drug approach in the treatment of some complex diseases. Drug Des Devel Ther 14:3235-3249. https://doi.org/10.2147/DDDT.S257494 
  32. Loscher W, Klein P (2022) New approaches for developing multi-targeted drug combinations for disease modification of complex brain disorders. Does epilepsy prevention become a realistic goal? Pharmacol Ther 229:107934. https://doi.org/10.1016/j.pharmthera.2021.107934 
  33. Alvarez XA, Alvarez I, Iglesias O, Crespo I, Figueroa J, Aleixandre M, Linares C, Granizo E, Garcia-Fantini M, Marey J (2016) Synergistic increase of serum BDNF in Alzheimer patients treated with cerebrolysin and donepezil: association with cognitive improvement in ApoE4 cases. Int J Neuropsychopharmacol 19:pyw024. https://doi.org/10.1093/ijnp/pyw024 
  34. Teng H, Li C, Zhang Y, Lu M, Chopp M, Zhang ZG, MelcherMourgas M, Fleckenstein B (2021) Therapeutic effect of Cerebrolysin on reducing impaired cerebral endothelial cell permeability. NeuroReport 32:359-366. https://doi.org/10.1097/WNR.0000000000001598 
  35. Tran L, Alvarez XA, Le H-A, Nguyen D-A, Le T, Nguyen N, Nguyen T, Nguyen T, Vo T, Tran T (2022) Clinical efficacy of cerebrolysin and cerebrolysin plus nootropics in the treatment of patients with acute ischemic stroke in Vietnam. CNS Neurol Disord Drug Targets 21:621-630. https://doi.org/10.2174/1871527320666210820091655 
  36. Rice-evans CA, Miller NJ, Bolwell PG, Bramley PM, Pridham JB (1995) The relative antioxidant activities of plant-derived polyphenolic flavonoids. Free Radic Res 22:375-383. https://doi.org/10.3109/10715769509145649 
  37. Wrobel-Biedrawa D, Grabowska K, Galanty A, Sobolewska D, Podolak I (2022) A flavonoid on the brain: quercetin as a potential therapeutic agent in central nervous system disorders. Life 12:591. https://doi.org/10.3390/life12040591 
  38. Oddo S, Caccamo A, Shepherd JD, Murphy MP, Golde TE, Kayed R, Metherate R, Mattson MP, Akbari Y, LaFerla FM (2003) Triple-transgenic model of Alzheimer's disease with plaques and tangles: intracellular Abeta and synaptic dysfunction. Neuron 39:409-421. https://doi.org/10.1016/s0896-6273(03)00434-3 
  39. Taylor MA, Khathayer F, Ray SK (2019) Quercetin and sodium butyrate synergistically increase apoptosis in rat C6 and human T98G glioblastoma cells through inhibition of autophagy. Neurochem Res 44:1715-1725. https://doi.org/10.1007/s11064-019-02802-8 
  40. Singh S, Kumar P (2018) Piperine in combination with quercetin halt 6-OHDA induced neurodegeneration in experimental rats: biochemical and neurochemical evidences. Neurosci Res 133:38-47. https://doi.org/10.1016/j.neures.2017.10.006 
  41. Ersoz M, Erdemir A, Derman S, Arasoglu T, Mansuroglu B (2020) Quercetin-loaded nanoparticles enhance cytotoxicity and antioxidant activity on C6 glioma cells. Pharm Dev Technol 25:757-766. https://doi.org/10.1080/10837450.2020.1740933 
  42. Raghu G, Berk M, Campochiaro PA, Jaeschke H, Marenzi G, Richeldi L, Wen FQ, Nicoletti F, Calverley PMA (2021) The multifaceted therapeutic role of N-acetylcysteine (NAC) in disorders characterized by oxidative stress. Curr Neuropharmacol 19:1202-1224. https://doi.org/10.2174/1570159X19666201230144109 
  43. Tardiolo G, Bramanti P, Mazzon E (2018) Overview on the effects of N-acetylcysteine in neurodegenerative diseases. Molecules 23:3305. https://doi.org/10.3390/molecules23123305 
  44. Huang Q, Aluise CD, Joshi G, Sultana R, St Clair DK, Markesbery WR, Butterfield DA (2010) Potential in vivo amelioration by N-acetyl-L-cysteine of oxidative stress in brain in human double mutant APP/PS-1 knock-in mice: toward therapeutic modulation of mild cognitive impairment. J Neurosci Res 88:2618-2629. https://doi.org/10.1002/jnr.22422 
  45. Pauletti A, Terrone G, Shekh-Ahmad T, Salamone A, Ravizza T, Rizzi M, Pastore A, Pascente R, Liang LP, Villa BR, Balosso S, Abramov AY, van Vliet EA, Del Giudice E, Aronica E, Patel M, Walker MC, Vezzani A (2019) Targeting oxidative stress improves disease outcomes in a rat model of acquired epilepsy. Brain 142:e39. https://doi.org/10.1093/brain/awz130 
  46. Hara J, Shankle WR, Barrentine LW, Curole MV (2016) Novel therapy of hyperhomocysteinemia in mild cognitive impairment, Alzheimer's Disease, and other dementing disorders. J Nutr Health Aging 20:825-834. https://doi.org/10.1007/s12603-016-0688-z 
  47. Kim D (2016) Guideline for the drug master file (DMF) of oriental (herbal) medicine preparation (# C0-2016-3-001). Ministry of Food and Drug Safety, Republic of Korea. https://dl.nanet.go.kr/SearchDetailView.do?cn=MONO1201633791#none 
  48. EVER-Pharma (2022) How to use Cerebrolysin®. EVER Pharma. https://www.cerebrolysin.com/cerebrolysin/how-to-use-cerebrolysin. Accessed 11 August 2022 
  49. Campanella B, Bramanti E (2014) Detection of proteins by hyphenated techniques with endogenous metal tags and metal chemical labelling. Analyst 139:4124-4153. https://doi.org/10.1039/c4an00722k 
  50. Codorniu-Hernandez E, Mesa-Ibirico A, Hernandez-Santiesteban R, Montero-Cabrera LA, Martinez-Luzardo F, Santana-Romero JL, Borrmann T, Stohrer WD (2005) Essential amino acids interacting with flavonoids: a theoretical approach. Int J Quant Chem 103:82-104. https://doi.org/10.1002/qua.20391 
  51. Boese AD, Codorniu-Hernandez E (2012) Cross-talk between amino acid residues and flavonoid derivatives: insights into their chemical recognition. Phys Chem Chem Phys 14:15682-15692. https://doi.org/10.1039/C2CP42174G 
  52. Balsamo R, Lanata L, Egan CG (2010) Mucoactive drugs. Eur Respir Rev 19:127-133. https://doi.org/10.1183/09059180.00003510 
  53. Pedre B, Barayeu U, Ezerina D, Dick TP (2021) The mechanism of action of N-acetylcysteine (NAC): the emerging role of H2S and sulfane sulfur species. Pharmacol Ther 228:107916. https://doi.org/10.1016/j.pharmthera.2021.107916