RESUMEN
Clinopodium nepeta subsp. spruneri is an aromatic herb with a mint-oregano flavor, used in Mediterranean regions in traditional medicine. The aerial parts of the plant are rich in essential oil that has antioxidant, antimicrobial and anti-inflammatory properties as well as insecticidal activity. The aim of our work was to determine the yield and composition of the essential oil of the plant, in relation to the harvest season and cultivation method, i.e., outdoor, greenhouse and in vitro culture, using gas chromatography-mass spectrometry (GC-MS) as an analytical tool. Essential oil yield fluctuated similarly in outdoor and greenhouse plants during the year (0.9-2.6%), with higher percentages (2.1-2.6%) in the hottest periods June-October (flowering stage) and April (vegetative stage), and was similar to the yield in in vitro plants (1.7%). More compounds were identified in the oil of outdoor and greenhouse plants (35) compared to that of in vitro plants (21), while the main compounds were the same, i.e., pulegone (13.0-32.0%, highest in February-April, 15.0% in vitro), piperitenone oxide (3.8-31.8%, lowest in February, 34.2% in vitro), piperitone epoxide (4.6-16.4%, highest in February, 15.5% in vitro), D-limonene (2.1-8.8%, lowest in February, 10.0% in vitro), isomenthone (2.3-23.0%, highest in February, 4.6% in vitro), germacrene D (1.9-6.5% highest in December-April, 2.9% in vitro) and dicyclogermacrene (2.1-5.3%, highest in December-April, 5.2% in vitro). Therefore, greenhouse and in vitro cultures were equally efficient in yielding essential oil and its constituents as outdoor cultivation, while in outdoor and greenhouse cultivations, the harvest season, mainly due to the prevailing ambient temperatures, affected the essential oil yield and its percentage composition.
RESUMEN
INTRODUCTION: We have previously shown that estrogen administered in ovariectomized female rabbits significantly reduce myocardial infarct size. We now investigated whether the phytoestrogen genistein similarly protects ischemic myocardium and whether this is associated with its antioxidant properties. In addition, we examined whether genistein abolishes preconditioning, since at high doses, it inhibits tyrosine kinase. MATERIALS AND METHODS: We studied five groups of New Zealand white female rabbits. Group A (n = 12) were normal controls, group B (n = 14) were ovariectomized 4 weeks prior to the experiment, group C (n = 10) were ovariectomized and treated with genistein (0.2 mg kg(-1) day(-1) subcutaneously) for 4 weeks before the experiment, group D (n = 12) had intact gonads and were treated with genistein (0.2 mg kg(-1) day(-1) subcutaneously) for 4 weeks before the experiment and group E (n = 8) were ovariectomized 4 weeks prior to the experiment and treated with a single dose of genistein (0.2 mg kg(-1) day(-1) subcutaneously) just prior to the experiment. All animals underwent 30 min of heart ischemia and 120 min of reperfusion, with (subgroup I) or without (subgroup II) preconditioning. Malondialdehyde (MDA) concentration just before the experiment was determined. RESULTS: We found significant differences between the groups-p < 0.0001 in factorial ANOVA. The groups with preconditioning had significant smaller infarcts compared to those without-AI vs AII (10.66 +/- 1.42% vs 43.22 +/- 2.67%), BI vs BII (18.53 +/- 2.36% vs 43.05 +/- 8.37%), CI vs CII (10.17 +/- 2.07% vs 44.5 +/- 5.47%), DI vs DII (14.98 +/- 2.36% vs 37.79 +/- 3.92%) and EI vs EII (17.11 +/- 3.24% vs 42.08 +/- 3.42%), p < 0.0005. Ovariectomy was not associated with larger myocardial infarctions-AII vs BII, p = NS. Genistein, for 4 weeks, did not protect ischemic myocardium in either ovariectomized or non-ovariectomized animals-BII vs CII and AII vs DII, p = NS. There was no significant difference between the preconditioned animals, with intact gonads or ovariectomized (AI vs BI, p = NS), ovariectomized with or without genistein (BI vs CI, p = NS) and non-ovariectomized whether treated with genistein or not (AI vs DI, p = NS). A single dose of genistein did not offer any protection (EII vs BII, p = NS), nor did it modify the preconditioning effect (EI vs BI, p = NS). We found no significant difference in MDA plasma levels between the groups. CONCLUSION: Genistein, at this dose, does not reduce infarct size per se nor abolishes the protection induced by preconditioning, in both ovariectomized and non-ovariectomized animals. Preconditioning offers myocardial protection in animals with intact gonads as well as estrogen deprived; bilateral ovariectomy, at least during short-term, is not associated with larger myocardial infarcts compared to control animals. In addition estrogen deprivation, during short term, as well as genistein do not modify oxidative stress.
