Extracts of Artemisia ciniformis Protect Cytotoxicity Induced by Hydrogen Peroxide in H9c2 Cardiac Muscle Cells through the Inhibition of Reactive Oxygen Species
Objective. Artemisia ciniformis (Asteraceae) and A. biennis are two of 34 Artemisia species growing naturally in Iran. In this study we investigated whether different extracts of A. ciniformis and A. biennis have protective effect against hydrogen peroxide-induced cytotoxicity in rat cardiomyoblast cells (H9c2). Method. The dried and ground aerial parts of these two species were extracted successively using petroleum ether (40–60), dichloromethane, ethyl acetate (EA), ethanol (EtOH) and ethanol？:？water (1？:？1) by maceration method. To evaluate whether different extracts of A. ciniformis and A. biennis protect cardiomyoblast H9c2 cells from H2O2 cytotoxicity, we examined the direct cytotoxic effect of H2O2 on H9c2 cells in the presence and absence of different extracts. After then, cell viability was measured by MTT assay. Results. H2O2 induced cytotoxicity in a concentration dependent manner. The IC50 value was 62.5？μM for 24？h exposure. However, pretreatment of cells with various concentrations of EA, EtOH, and EtOH/wt extract of A. ciniformis protected cells from H2O2-induced cytotoxicity. Moreover, pretreatment with EA, EtOH and EtOH/wt extracts of A. ciniformis lead to a decrease in the reactive oxygen species (ROS) generation. Taken together our observation indicated that nontoxic concentration of different extracts of A. ciniformis has protective effect on H2O2-induced cytotoxicity in H9c2 cells. 1. Introduction Artemisia biennis Willd. and A. ciniformis Krasch. & Popov ex Poljakov. (Compositae) grow wildly in Iran [1]. Analysis of the essential oils from the aerial parts of A. biennis growing in Iran and western Canada revealed the presence of camphor and [E] beta-farnesene as the major constituents, respectively [2, 3]. Myrcene [4] and davanone [5] have been reported as the main constituent in the aerial parts oils of A. ciniformis Cytotoxicity of some fractions of A. biennis and A. ciniformis as well as significant effects of ethanolic extracts of the species on in vitro leishmanicidal activity have been proved [6–8]. Iranshahi et al. [9] reported the presence of high amounts of sesquiterpene lactonesin A. ciniformis. Another study showed that antioxidant activity and total phenolic content of hydroethanolic extract of A. biennis were higher than those of other extracts [10]. Oxidative stress corresponds to an imbalance between the rate of oxidant production and degradation. It causes numerous biological effects ranging from alternation in signal transduction and gene expression to mutagenesis and finally cell death. It is well known that
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