EVALUATION OF ANTIAPOPTOTIC AND PROTECTIVE EFFECTS OF JUGLANS REGIA ESSENTIAL OIL ON THE TREATMENT OF TRAMADOL IN PC12 CELLS

Background and Aim : Tramadol is an opioid pain reliever that its abuse may lead to neurotoxicity in the brain cells. In high concentration, it can induces cell death through increasing the cell cytotoxicity. It increases the production of nitric oxide, activates the caspases, and decreases the mitochondrial membrane potential in central nervous system. In this study, we aimed to investigation of suppressing effects of Juglans regia oil on tramadol-induced cell death. Methods : In this study, we used the rat neuron-like cell line, PC12. Trypan blue test was performed for assessment of cell viability. Cell proliferation assessed by MTT test and cell cytotoxicity measured through LDH test. TUNEL test clarified the DNA fragmentation and apoptosis occurrence. Griess reaction was used to measurement of NO production and Rat V-Plex Kit was used to measurement of IL-1β, IL-6, INFγ, and TNFα. Rhodamine-123 and Caspase-3 Colorimetric Assay Kit were used to measurement of mitochondrial membrane potential and caspase-3 activity, respectively.Results : The viability and proliferation percent of all the Juglans regia oil treatments were higher than tramadol-treated cell by significant intracellular difference. The cell cytotoxicity and cell death index of all the Juglans regia oil treatments were lower than tramadol-treated cell. All treatment increased mitochondrial membrane potential and decreased NO production, IL-1 , IL-6, INF , and TNF release, and caspase-3 activity.Conclusion : we concluded that Juglans regia oil suppressed the tramadol-induced cell death in neuron-like, PC12 cells through inhibition of NO production, mitochondrial membraine disruption, DNA fragmentation, and caspase-3 activity

Keywords:

Juglans regia oil , tramadol , cell death , apoptosis , cytotoxicity