Investigating Quantitative analysis of the gene expression of calcium/calmodulin-dependent protein kinase IV by the effect of Olibanum alcoholic extract in PC12 cell line

Abstract

Background & Objective: Long-term memory depends on protein synthesis. The product of Camkiv gene promotes memory via activating its proteins. The treatment of laboratory animals by Olibanum leads to memory improvement and the recovery of Alzheimer. Therefore, the aim of this study is the evaluation of Olibanum ethanolic extract on the Camkiv expression in PC12 cells. Materials & Methods: Olibanum toxicity on the cell viability was investigated by MTT test. Cells were treated with concentrations 10,25,40,55,70 and 85 μg/ml of extract in time intervals 12,24,48 and 72 hours and their absorption rate was measured. Then, cells were treated by concentrations 2 and 20 μg/ml of extract in mentioned times. Extracted RNA was converted into cDNA and real-time PCR performed. Results: Cell death was raised by increasing time and concentration of extract treatment. IC50 values were obtained as 71.01, 52.95, 21.05 and 13.85 μg/ml in 12, 24, 48 and 72 hours of treatment, respectively. Besides, concentrations 2 and 20 μg/ml significantly increased Camkiv expression following 24 hour treatment. The maximum expression of Camkiv was observed in 48 hour treatment. The effect of Olibanum on gene upregulation was stable until 72 hours. Conclusion: The Olibanum ethanolic extract can remarkably upregulate Camkiv expression for a long time. These results are consistent with the previous studies indicating the effect of Olibanum on upregulation of Bdnf, Camkiv‌-downstream gene. However, regarding the existence of the two-direction pathway in the expression regulation of Bdnf and Camkiv‌, comprehensive studies are required to determine exact mechanism of Olibanum function in the brain