RESUMEN
In the present study, we assessed the antioxidant and antileishmanial potential from fresh leaves of Ballota (B.) hirsuta essential oil (EO). The GC-MS analysis of B. hirsuta EO revealed that spathulenol and germacrene D were the main components accounting for 26.03% and 19.64% of the total EO, respectively. B. hirsuta EO possesses moderate antioxidant activity, both in neutralizing DPPH radicals and in inhibiting ß-carotene bleaching. In addition, it exhibits both high antileishmanial activity and selectivity towards the promastigote and amastigote forms. Specifically, B. hirsuta EO showed an IC50 value of 20.78 µg/mL and 23.62 µg/mL, against the promastigote and amastigote forms of L. infantum, respectively. It also demonstrated an IC50 value of 22.39 and 25.76 µg/mL, against the promastigote and amastigote forms of L. major, respectively. However, it exhibited moderate cytotoxicity, with a selectivity index below 10. The investigation of the molecular mechanism of action revealed that B. hirsuta EO inhibited only the sterol pathway, including CYP51 gene expression. Additionally, in silico analysis indicated that the main compounds of B. hirsuta EO, germacrene and spathulenol, exhibited excellent affinity energy against Leishmania enzymes trypanothione reductase (TryR) and trypanothione synthase (TryS). This denotes the potential of these compounds as promising agents to control leishmaniasis.
RESUMEN
The present study aimed to evaluate the leishmanicidal potential of the essential oil (EO) of Micromeria (M.) nervosa and to investigate its molecular mechanism of action by qPCR. Furthermore, in silicointeraction study of the major M. nervosa EO compounds with the enzyme cytochrome P450 sterol 14α-demethylase (CYP51) was also performed. M. nervosa EO was analyzed by gas chromatography-mass spectrometry (GC-MS). Results showed that α-pinene (26.44%), t-cadinol (26.27%), caryophyllene Oxide (7.73 ± 1.04%), and α-Cadinene (3.79 ± 0.12%) are the major compounds of M. nervosa EO. However, limited antioxidant activity was observed, as this EO was ineffective in neutralizing DPPH free radicals and in inhibiting ß-carotene bleaching. Interestingly, it displayed effective leishmanicidal potential against promastigote (IC50 of 6.79 and 5.25 µg/mL) and amastigote (IC50 of 8.04 and 7.32 µg/mL) forms of leishmania (L.) infantum and L. major, respectively. Molecular mechanism investigation showed that M. nervosa EO displayed potent inhibition on the thiol regulatory pathway. Furthermore, a docking study of the main components of the EO with cytochrome P450 sterol 14α-demethylase (CYP51) enzyme revealed that t-cadinol exhibited the best binding energy values (-7.5 kcal/mol), followed by α-cadinene (-7.3 kcal/mol) and caryophyllene oxide (-7 kcal/mol). These values were notably higher than that of the conventional drug fluconazole showing weaker binding energy (-6.9 kcal/mol). These results suggest that M. nervosa EO could serve as a potent and promising candidate for the development of alternative antileishmanial agent in the treatment of leishmaniasis.
Asunto(s)
Antiprotozoarios , Simulación del Acoplamiento Molecular , Aceites Volátiles , Aceites Volátiles/farmacología , Aceites Volátiles/química , Antiprotozoarios/farmacología , Antiprotozoarios/química , Antioxidantes/farmacología , Antioxidantes/química , Cromatografía de Gases y Espectrometría de Masas , Esterol 14-Desmetilasa/metabolismo , Esterol 14-Desmetilasa/química , Simulación por Computador , Leishmania/efectos de los fármacos , Leishmania/enzimología , Monoterpenos Bicíclicos/farmacología , Monoterpenos Bicíclicos/químicaRESUMEN
AIMS: This study aimed to determine the antibacterial and antileishmanial potential of Micromeria nervosa extracts. The identification of the antileishmanial compound and the study of its molecular mechanism of action have also been undertaken. METHODS AND RESULTS: Ethanol extract showed high polyphenol content and diethyl ether extract exhibited high DPPH scavenging and low beta-carotene bleaching activity (IC50 = 13.04 ± 0.99 and 200.18 ± 3.32 µg mL-1, respectively). However, diethyl ether extract displayed high antibacterial activity against Gram-positive strains including methicillin-resistant Staphylococcus aureus (MIC = 31.25 µg mL-1), Staph. aureus ATCC6538 (MIC = 62.5 µg mL-1), and Listeria monocytogenes ATCC 19115 (MIC = 125 µg mL-1), as well as high antileishmanial activity against the promastigote forms of L. infantum and L. major (IC50 = 11.45 and 14.53 µg mL-1, respectively). The active compound was purified using bioassay-guided fractionation and thin layer chromatography, and identified as ursolic acid using high-performance liquid chromatography coupled with a photodiode array and mass spectrometry. The purified compound was strongly inhibitory against the promastigote and amastigote forms of L. infantum and L. major (IC50 = 5.87 and 6.95 µg mL-1 versus 9.56 and 10. 68 µg mL-1, respectively) without overt cytotoxicity against Raw 264.7 macrophage cells (SI = 13.53 and 11.43, respectively). The commercial compound (ursolic acid) showed similar activity against amastigotes and promastigotes forms of L. infantum and L. major. Moreover, its molecular mode of action against leishmaniasis seems to involve the expression of the ODC and SPS genes involved in thiol pathway. CONCLUSION: Extracts of M. nervosa can be considered as a potential alternative to antimicrobial and antileishmanial drugs.
Asunto(s)
Antiinfecciosos , Antiprotozoarios , Lamiaceae , Staphylococcus aureus Resistente a Meticilina , Antioxidantes/farmacología , Antioxidantes/análisis , Éter , Extractos Vegetales/farmacología , Extractos Vegetales/química , Antiprotozoarios/farmacología , Antibacterianos/farmacología , Antiinfecciosos/farmacología , Staphylococcus aureus , Ácido UrsólicoRESUMEN
Echium arenarium Guss is a Mediterranean plant traditionally used in healing skin wound and it was reported exhibiting potent antioxidant, antibacterial, and antiparasitic activities. However, antitumoral activities of this plant have not yet been explored. Here we investigated for the first time, root (EARE) and aerial part (EAAPE) extracts of E. arenarium Guss to examine cytotoxicity and apoptosis activation pathway on U266 human multiple myeloma (MM) cell line. We demonstrated that EARE and EAAPE decreased U266 cell viability in a dose dependent manner. Based on 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, EARE was significantly two times more efficient (IC50 value 41 µg/ml) than EAAPE (IC50 value 82 µg/ml) considering 48 h of treatment. Furthermore, after 24 h of exposure to 100 µg/ml of EARE or EAAPE, cell cycle showed remarkable increase in sub-G1 population and a decrease of U266 cells proportion in G1 phase. In addition, EARE increased cell percentage in S phase. Moreover, analysis revealed that EAAPE or EARE induced apoptosis of U266 cells after 24 h of treatment. Interestingly, depolarization of mitochondrial membrane potential and activation of caspase 3/7 were demonstrated in treated U266 cells. Phytochemical analysis of E. arenarium extracts showed that EARE exhibited the highest content of total phenolic content. Interestingly, six phenolic compounds were identified. Myricitrin was the major compound in EARE, followed by luteolin 7-O-glucoside, resorcinol, polydatin, Trans-hydroxycinnamic acid, and hyperoside. These findings proved that an intrinsic mitochondria-mediated apoptosis pathway probably mediated the apoptotic effects of E. arenarium Guss extracts on U266 cells, and this will suggest several action plans to treat MM.
Asunto(s)
Echium , Mieloma Múltiple , Apoptosis , Línea Celular Tumoral , Proliferación Celular , Humanos , Mitocondrias , Mieloma Múltiple/tratamiento farmacológico , Componentes Aéreos de las Plantas , Extractos VegetalesRESUMEN
The present work was developed to evaluate the in vitro antioxidant, antibacterial, antileishmanial and cytotoxic activities of Echium arenarium (Guss) extracts, and to analyze their phytochemical composition. The highest content of total phenolic compounds was obtained in the ethyl acetate extract which showed the best DPPH scavenging activity and ß-carotene bleaching inhibition (IC50â¯=â¯1.1 and 9.94⯵g/mL respectively). It also exhibited the highest antibacterial activity against Gram-positive bacteria (L. monocytogenes; S. aureus; MRSA, E. faecalis and B. cereus) and antileishmanial activity against L. major (IC50â¯=â¯13.91⯱â¯0.43 µg/mL) and L. infantum (IC50â¯=â¯9.91⯱â¯0.15 µg/mL). Moreover, the active extract exhibited potent antiamastigote activity (IC50â¯=â¯22.48⯱â¯0.14⯵g/mL and 18.59⯱â¯0.09⯵g/mL against L. major and L. infantum respectively). Cytotoxicity studies revealed low toxicity against Raw 264.7 macrophage cell line (IC50â¯=â¯145.80⯱â¯0.84 µg/mL, SIâ¯<â¯10). Luteolin-7-O-glucoside was identified as the major flavonoid component by RP-HPLC analysis. In conclusion, Echium arenarium (Guss) extract was characterized by a wide range of biological activities and could be used as a potential natural anti-infectious drug.
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Antiinfecciosos/farmacología , Echium/química , Fitoquímicos/química , Fitoquímicos/farmacología , Extractos Vegetales/química , Extractos Vegetales/farmacología , Animales , Antibacterianos/farmacología , Antiinfecciosos/química , Antioxidantes/análisis , Antioxidantes/farmacología , Compuestos de Bifenilo , Cromatografía Líquida de Alta Presión , Flavonas/análisis , Flavonoides/análisis , Glucósidos/análisis , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Grampositivas/efectos de los fármacos , Leishmania infantum/efectos de los fármacos , Leishmania major/efectos de los fármacos , Ratones , Pruebas de Sensibilidad Microbiana , Fenoles/análisis , Fenoles/farmacología , Picratos , Células RAW 264.7/efectos de los fármacos , beta CarotenoRESUMEN
The effects of two extraction methods, used at three ripening stages on the total polyphenol contents and the antioxidant activities of Cumin (Cuminum cyminum L.) seed extracts were studied. The ripening stage effect on some physical properties of cumin seed was significant. The increase of dry matter (from 10.3 to 87.5%) during ripeness was correlated negatively with that of moisture content (from 89.7 to 12.5%). Besides results showed that the full ripe seeds were richer on polyphenols and condensed tannin than unripe ones, and consequently exhibited higher antioxidant activities. However, the unripe seeds had a higher total flavonoid content compared to those of half ripe and full ripe ones. The comparison of two extraction methods showed that soxhlet extracts contained the greatest amount of polyphenols and flavonoids, while maceration samples exhibited higher antiradical and bleaching power assay. Total phenolic contents and IC50 (concentration required to cause a 50% DPPH inhibition) values in cumin seed during their maturation allowed to conclude that antioxidant activity does not depend only on the high content of total phenolics but also on the phenolic composition. A total of 19 phenolic compounds were successfully identified by HPLC analysis during the ripening of cumin seeds. Rosmarinic acid was the major phenolic acid for the unripe seeds. Furthermore, half ripe and full ripe seeds were dominated by p-coumaric acid. These compounds might be considered as interesting bioactive natural substances that may be used in several fields, such as nutraceuticals, cosmetics and agro-food industry.
Asunto(s)
Antioxidantes/farmacología , Cuminum/química , Extractos Vegetales/farmacología , Polifenoles/farmacología , Semillas/química , Especias/análisis , Antioxidantes/análisis , Compuestos de Bifenilo/metabolismo , Cinamatos/análisis , Cinamatos/farmacología , Ácidos Cumáricos/análisis , Ácidos Cumáricos/farmacología , Depsidos/análisis , Depsidos/farmacología , Flavonoides/análisis , Flavonoides/farmacología , Humanos , Fenoles/análisis , Fenoles/farmacología , Picratos/metabolismo , Extractos Vegetales/análisis , Polifenoles/análisis , Propionatos , Taninos/análisis , Agua/análisis , Ácido RosmarínicoRESUMEN
Two Carthamus tinctorius varieties (Jawhara and 104) were studied in order to investigate their natural dyes contents and biological activities. Obtained results showed that quinochalcone contents and antioxidant activities varied considerably as function of flowering stages. So flowers at fructification stage contained the highest carthamin content with the strongest antioxidant capacity with all assays (FRAP, DPPH, and chelating power methods). In parallel, we showed a decrease in the content of precarthamin. The quantitative variation of these molecules could be due to colour change of C. tinctorius flowers. Correlation analysis indicated that the ABTS method showed the highest correlation coefficients with carthamin and precarthamin contents, that is, 0.886 and 0.973, respectively. Concerning the regional effect, the contents of precarthamin and carthamin varied significantly (P < 0.05) at studied regions with the optimum production given by samples of Beja (902.41 µg/g DW and 42.05 µg/g DW, respectively, at flowering stage). During flowering, the antimicrobial activity of these two natural dyes increased where the maximum inhibitory effect mentioned with carthamin mainly against E. coli (iz = 25.89 mm) at fructification stage. Therefore, the increased frequency of resistance to commonly used antibiotics leads to the search for new effective natural drugs at food and pharmaceutical industries.