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Protective effects of aqueous and ethanol extracts of rosemary on H2O2-induced oxidative DNA damage in human lymphocytes by comet assay

Abstract Background: Rosemary (Rosmarinus officinalis) possesses various pharmacological properties such as antioxidant, anti-tumorigenesis and anti-mutagenesis activities. In this study, we investigated the possible protective effects of ethanol and aqueous extracts of rosemary on human lymphocyte DNA damage induced by H2O2. The extent of DNA lesions was measured using comet assay. Methods: Blood samples were taken from healthy volunteers and lymphocytes were isolated. The lymphocytes were then incubated in aqueous and ethanol extract of rosemary (0.05, 0.1, 0.5, 1 and 2.5 mg/mL) and H2O2 (50, 100 and 200 mM). Lymphocytes were also incubated with a combination of H2O2 (100 mM) with either 1 or 2.5 mg/mL of both extracts for 30 min at 4°C. Results: Our findings showed that H2O2 treatment led to a significant concentrate-dependent DNA damage in human lymphocyte when compared to respective controls (p<0.001). The DNA damage which was initially occurred as the result of 100 µM H2O2 (Percentage tail DNA 55.1%) was inhibited due to the ethanol extract of rosemary at the doses tested (percentage tail DNA 4.7% and 4.03%). However, the aqueous extract has no effects on H2O2 genotoxicity. Conclusions: We suggest that antioxidant constituents in ethanol extract of rosemary can prevent human lymphocytes oxidative DNA damage which is due to its free radical scavenging activity.