Antioxidant Effects of Pinus eldarica Extracts Against Cisplatin-induced Cytotoxicity in Human Hepatoma (HepG2) Cell Line

Abstract

Background: Cisplatin has cytotoxic effects through free radical generation. Reports indicate that various members of Pinaceae family may have antioxidant properties. In this study we investigated the cytoprotective effects of needle volatile oil and bark extract of Pinus eldarica in Hepatoma G2 (HepG2) cell line. Methods: Using the maceration technique, we obtained the ethanolic extract of Pinus eldarica’s bark (BHAEPE). Folin-ciocalteu reagent was used to determine the total phenolic content of BHAEPE. The Needle’s Volatile Oil of Pinus eldarica (NVOPE) was obtained by Clevenger hydrodistillation method. The main components of NVOPE were identified by Gas Chromatography and Mass Spectrophotometry (GC-MS). We used DPPH assay to investigate the antioxidant activity of both NVOPE and BHAEPE. Also, MTT assay was performed to test the protective effects of both BHAEPE and NVOPE against cisplatin. Results: Folin-ciocalteu test demonstrated that each gram of BHAEPE was equivalent to 371±6 mg of gallic acid. Also, GC-MS identified germacrene D as the main component of NVOPE. BHAEPE had more antioxidant capacity compared to NVOPE. When incubated solely with cells, neither BHAEPE nor NVOPE represented cytotoxicity on. Furthermore, BHAEPE demonstrated mitogenic effects on higher doses (50, 75 and 100 μg/ml). However, both NVOPE and lower doses of BHAEPE (10, 20 μg/ml) could not protect the cells against cisplatin’s cytotoxicity, but higher doses of BHAEPE provided cytoprotective effects. Conclusion: The antioxidant properties of both NVOPE and lower doses of BHAEPE did not adequately protect HepG2 cells against the cisplatin’s cytotoxicity. However, BHAEPE at high concentrations demonstrated cytoprotective effects.

Keywords

• pinus eldarica
• cisplatin
• dpph test
• mtt assay
• hepg2