020P London, UK
8th European Workshop on Cannabinoid Research



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.


(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 J23: 3020-3029.

(4) Ma L et al. (2015) Sci Rep 5: 12440.

(5) Noonan J et al. (2010). J Biol Chem 285: 38543-38554.