THE EFFECT MATERIAL (PRUNUS AMYGDALUS VAR AMARA) AND DIFFERENT OIL CONCENTRATIONS ON MORPHINE INDUCED CELL DEATH IN PC12

Background and Aim : High concentration of morphine induces cell death through increase the cell cytotoxicity, production of Nitric oxide, inflammatory cytokines, and caspase-3 in central nervous system (CNS). Purpose of this research, is study of inhibitory effects of Prunus amygdalus var amara essential oil on morphine-induced cell death in neuron-like cells, PC12.Methods : The Gas Chromatography Mass Spectroscopy GC-MS used to chemical analysis of Prunus amygdalus var amara essential oil. The cell viability, cell proliferation, and cell cytotoxicity were analyzed by Trypan blue, MTT and Lactate dehydrogenase (LDH) tests, respectively. DNA fragmentation detected by Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) test. NO production measured through Griess reaction. Inflammatory cytokines of IL-1β, IL-6, INFγ, and TNFα were analyzed by Rat V-Plex Kit. Mitochondrial membrane potential detected by rhodamine-123 and caspase-3 activity observed through Caspase-3 Colorimetric Assay Kit.Results : GC-MS indicated that Benzaldehyde and Benzoic acid are the most frequently found chemical constituents in Prunus amygdalus var amara essential oil. All Prunus amygdalus var essential oil treatments shown higher proliferation and viability and lower cytotoxicity and cell death index compared to morphine-treated cells. Production of NO, IL-1β, IL-6, INFγ, and TNFα release, and Caspase-3 were decreased in presence of Prunus amygdalus var amara essential oil while mitochondrial membrane potential decreased.Conclusion : We concluded that Prunus amygdalus var amara essential oil suppressed the cell death which induced by morphine in PC12 cells. It can inhibits the production of NO. Also, it can inhibit apoptosis through inhibition of caspase-3 activity, DNA fragmentation, and disruption of mitochondrial membrane

Keywords:

Morphine , cell death , apoptosis , Prunus amygdalus var amara essential oil