Satureja subspicata and S. horvatii Extracts Induce Overexpression of the BCl-2 Family of Anti-apoptotic Genes and Reduce Micronuclei Frequency in Mice

Abstract

Satureja subspicata and S. horvatii are endemic species of the Balkan Peninsula and often used in traditional medicine in Bosnia and Herzegovina to treat different health conditions. We aimed to analyze the unevaluated apoptotic, genotoxic and cytotoxic effects of two Satureja species, as well as their content of phenolics that are mainly responsible for the plant's biological activity. Apoptotic and geno/cytotoxic activities of S. subspicata and S. horvatii were investigated in vitro in human lymphocyte culture and in vivo in mice. The content of the main phenolics in plant extracts was determined by ultra-high pressure liquid chromatography-MS-MS (UHPLC–MS/MS). Genotoxic and cytotoxic activities of Satureja extracts were evaluated in vitro by applying a cytokinesis-block micronucleus cytome assay in human lymphocyte culture and in vivo applying a mice reticulocytes micronucleus assay. SALSA RT-MLPA R011-C1 apoptosis assay was used for measuring the relative expression of 44 genes associated with the regulation of the apoptotic pathways in human lymphocyte cultures treated with different concentrations of two Satureja extracts. The first analysis of phenolic compounds in S. horvatii and S. subspicata determined by an UHPLC-MS/MS method revealed high levels of rosmarinic and caffeic acids. Minor genotoxic potential was determined in relation to the tested concentrations while no cytostatic and cytotoxic effects were revealed in vitro. However, when applied in concentrations of 200 mg/kg per os, aqueous extracts of two Satureja species significantly decreased frequency of reticulocytes micronuclei in treated mice against controls. Extracts of S. subspicata and S. horvatii in concentrations of 0.2 mg/mL, regardless of solvent used, downregulated pro-apoptotic and upregulated anti-apoptotic genes, showing anti-apoptotic activity. Our results indicate that the registered anti-genotoxic and anti-apoptotic activity is most likely related to the high level of phenolic acids (particularly rosmarinic and caffeic) in the tested extracts.

Bioactivity and phenolics profile of aqueous and ethyl acetate extracts of Satureja kitaibelii Wierzb. ex Heuff

“…increased Bcl-2/Bax ratio, was observed for methanol extract of S. hortensis, which was characterized by high amount of rosmarinic acis 66 . These data, together with previously con rmed low genotoxic potential of aqueous extracts of Satureja plants and rosmarinic acid as their dominant compound 31 , t well with safe usage of S. kitaibelii aqueous extracts, but this certainly needs to be better justi ed in future.…”

“…In accordance with our results on the cytotoxicity of ethyl acetate extracts, in a previous study, methanol extract of S. kitaibellii exhibited moderate activity against estrogen-dependant (MDA-MB-361) and estrogen-nondependant (MDA-MB-453) breast cancer, colon cancer (LS174), and HeLa cell lines, as well as strong cytotoxic activity against human malignant melanoma cells (Fem-x), which was partly correlated with tested activity of rosmarinic acid 3 . On the other hand, low cytotoxic potential of aqueous extracts of related species, S. subspicata and S. horvatii, was noted previously in human lymphocytes in vitro 31 , as well as of S. montana aqueous and aqueous ethanol extracts with IC 50 values above 600 μg/ml against Caco-2, TR146 and HeLa cell lines 36 . Moreover, aqueous and DMSO extracts of S. subspicata and S. horvatii downregulated pro-apoptotic and upregulated anti-apoptotic genes of the Bcl-2, showing anti-apoptotic activity, which authors linked with previously shown similar effects of rosmarinic acid.…”

“…Derivatives of HCA are common for plants belonging to the family Lamiaceae. Rosmarinic acid, is often one of the major polyphenol constituents in aerial parts polar extracts of these plants, including plants of the genus Satureja [28][29][30][31][32] . Besides rosmarinic acid, Lamiaceae species produce higher HCA oligomers which contain up to 8 monomer units 33,27 .…”

Bioactivity and phenolics profile of aqueous and ethyl acetate extracts of Satureja kitaibelii Wierzb. ex Heuff. obtained by ultrasound-assisted extraction

“…Derivatives of HCA are common for plants belonging to the family Lamiaceae. Rosmarinic acid is often one of the major polyphenol constituents in aerial parts of polar extracts of these plants, including plants of the genus Satureja [34][35][36][37][38] . Besides rosmarinic acid, Lamiaceae species produce higher HCA oligomers which contain up to 8 monomer units 33,39 .…”

Biological and Chemical Comparison of Natural and Cultivated Samples of Satureja macrantha C.A.Mey.

“…There is no study on the cytotoxicity of this species in the literature. However, many studies showed that other Satureja species had cytotoxic potential [41][42][43].…”