Abstract
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•Cupressus sempervirens extracts activities varied with phenological stage.•Methanolic extract from vegetative stage showed the highest antioxidant activity.•Ethyl-acetate extract was the most active against bacteria and Leishmania strains.•Cupressuflavone and amentoflavone were the major compounds identified.
Cupressus sempervirens L. is largely used in traditional medicine as an antimicrobial agent. The present study investigated the antioxidant, antibacterial and antileishmanial activities of C. sempervirens organic extracts at different phenological stages. Antioxidant activity was determined by DPPH (2,2-diphenyl-1-picrylhydrazylradical) scavenging assay, ferric reducing power and total antioxidant capacity. The antibacterial activity was evaluated against five clinical strains by disk diffusion method and minimum inhibitory concentration (MIC). The antileishmanial activity was determined against promastigote and amastigote forms of Leishmania (L.) infantum and L. major. Results of antioxidant activity showed that methanolic extract from vegetative stage had the most important activity. The ethyl acetate extract of C. sempervirens from flowering stage was the most active against Bacillus cereus (ATCC 14579) and Staphylococcus aureus (ATCC 25923) with MIC of 100 μg/mL and 50 μg/mL, respectively. Interestingly, this extract exhibited high antileishmanial activity against promastigote form of L. infantum and L. major (IC50 = 1.47 and 2.8 μg/mL, respectively) and amastigote form (IC50 = 3.61 and 5.42 μg/mL, respectively). Furthermore, ethyl acetate extract showed low cytotoxicity on macrophage cells Raw264.7 with selectivity index of 34.15 and 17.93 for L. infantum and L. major, respectively. The identification by HPLC and HPLC-MSn of active extracts of C. sempervirens revealed that major compounds of methanolic extract from vegetative stage and ethyl acetate extract from flowering stage were cupressuflavone and amentoflavone. Based on these results, C. sempervirens extracts could be used as an alternative to chemical drugs for the treatment of oxidative stress and infectious diseases.