Martin, S

Milton, NG

Paterson, A

Werner, S

Leeds Beckett Repository

Proceedings of the British Pharmacological Society at http://www.pA2online.org/abstracts/Vol17Issue1abst020P.pdf  Effect of RVD-hemopressin on amyloid-β induced toxicity in human SH-SY5Y neuroblastoma cells  S. J. Werner, S. W. Martin, A. W. Paterson, N. G. Milton. School of Clinical & Applied Sciences, Leeds Beckett University, Leeds, United Kingdom.  Introduction: Previous in vitro and in vivo studies have demonstrated the protective properties of lipid endocannabinoids against amyloid-β (Aβ) induced neurotoxicity1,2. Lipid-derived endocannabinoid agonists such as 2-arachidonoylglycerol (2-AG) can exert their effects via both the extra- and intracellular cannabinoid receptor-1 (CB1)3. Pepcans are a group of haemoglobin derived peptide cannabinoids and are found throughout the CNS3. They are cell-impermeant and act on the extracellular CB1 receptor3,4 as agonists/antagonists. The pepcan RVD-hemopressin (RVD) is a CB1 receptor agonist3. The aim of this study was to determine whether RVD is protective against Aβ toxicity. Method: This study employed MTT cell viability assays to investigate the effects of the peptide CB1 agonist RVD and lipid CB1 agonist 2-AG plus the CB1 antagonist AM281 on Aβ 25-35 induced neurotoxicity in human neuroblastoma SH-SY5Y cells. Data was analyzed by one-way analysis of variance (ANOVA). Results: RVD (0.01-10µM) had no effect on 10µM Aβ 25-35 induced neurotoxicity in SH-SY5Y cells, whereas 2-AG (0.02-10µM; P<0.05 vs Aβ 25-35 alone) promoted a concentration dependent inhibition (Figure 1A). The CB1 antagonist AM281 (10µM) had no effect on RVD (10µM) plus 10µM Aβ 25-35, however it abolished the protective effects of 2-AG (10µM; P<0.05 vs Aβ 25-35 alone) on 10µM Aβ 25-35 induced neurotoxicity (Figure 1B).   Figure 1. (A) Dose-response curves for RVD plus 10µM Aβ 25-35 and 2-AG plus 10µM Aβ 25-35 on MTT reduction in SH-SY5Y cells. (B) SH-SY5Y cells were exposed to 10µM Aβ 25-35 alone, or plus 10µM RVD alone or 10µM RVD and 10µM AM281 or 10µM 2-AG alone or 10µM 2-AG and 10µM AM281 and cell viability determined by MTT reduction. Results are mean ± SEM (n=8 for each data point); * = P< 0.05 vs Aβ 25-35 alone; † = P<0.05 vs control; (one-way ANOVA). Conclusion: In conclusion, the peptide cannabinoid RVD is non-protective against Aβ 25-35 induced neurotoxicity in SH-SY5Y cells. Lipid based endocannabinoids, such as 2-AG, are protective against Aβ 25-35 induced neurotoxicity1. Our results support the suggestion that endocannabinoid neuroprotection against Aβ involves the intracellular CB1 receptor rather than the extracellular CB1 receptor 5. References: (1) Milton NGN (2002). Neurosci Letts 332: 127-130. (2) van der Stelt M et al. (2006). Cell Mol Life Sci 63: 1410-1424. (3) Gomes I et al. (2009). FASEB J. 23: 3020-3029. (4) Ma L et al. (2015) Sci Rep 5: 12440. (5) Noonan J et al. (2010). J Biol Chem 285: 38543-38554.  

Effects of RVD-hemopressin on amyloid-β induced toxicity in SH-SY5Y neuronal cell culture

http://eprints.leedsbeckett.ac.uk/4986/1/vol17issue1abst020p.pdf

Effects of RVD-hemopressin on amyloid-β induced toxicity in SH-SY5Y neuronal cell culture

Abstract

Similar works

Full text

Available Versions

Leeds Beckett Repository