ABSTRACT
BACKGROUND: In the past two decades, the impacts of Helium-Neon (He-Ne) laser on stress resistance and secondary metabolism in plants have been studied, but the signaling pathway which by laser regulates this process remains unclear. Therefore, the current study sought to explore the role of RBOH-dependent signaling in He-Ne laser-induced salt tolerance and elicitation of secondary metabolism in Salvia officinalis. Seeds were primed with He-Ne laser (6 J cm- 2) and peroxide hydrogen (H2O2, 5 mM) and 15-old-day plants were exposed to two salinity levels (0, 75 mM NaCl). RESULTS: Salt stress reduced growth parameters, chlorophyll content and relative water content (RWC) and increased malodialdehyde (MDA) and H2O2 contents in leaves of 45-old-day plants. After 48 h of salt exposure, higher transcription levels of RBOH (encoding NADPH oxidase), PAL (phenylalanine ammonia-lyase), and RAS (rosmarinic acid synthase) were recorded in leaves of plants grown from seeds primed with He-Ne laser and/or H2O2. Despite laser up-regulated RBOH gene in the early hours of exposing to salinity, H2O2 and MDA contents were lower in leaves of these plants after 30 days. Seed pretreatment with He-Ne laser and/or H2O2 augmented the accumulation of anthocyanins, total phenol, carnasol, and rosmarinic acid and increased total antioxidant capacity under non-saline and more extensively at saline conditions. Indeed, these treatments improved RWC, and K+/Na+ ratio, enhanced the activities of superoxide dismutase and ascorbate peroxidase and proline accumulation, and significantly decreased membrane injury and H2O2 content in leaves of 45-old-day plants under salt stress. However, applying diphenylene iodonium (DPI as an inhibitor of NADPH oxidase) and N, N-dimethyl thiourea (DMTU as a H2O2 scavenger) after laser priming reversed the aforementioned effects which in turn resulted in the loss of laser-induced salt tolerance and secondary metabolism. CONCLUSIONS: These findings for the first time deciphered that laser can induce a transient RBOH-dependent H2O2 burst, which might act as a downstream signal to promote secondary metabolism and salt stress alleviation in S. officinalis plants.
Subject(s)
Cinnamates , Depsides , Rosmarinic Acid , Salt Tolerance , Salvia officinalis , Signal Transduction , Salvia officinalis/metabolism , Salvia officinalis/physiology , Salvia officinalis/drug effects , Salvia officinalis/genetics , Depsides/metabolism , Cinnamates/metabolism , Abietanes/metabolism , Hydrogen Peroxide/metabolism , Lasers , Plant Proteins/metabolism , Plant Proteins/genetics , Plant Leaves/metabolism , Plant Leaves/drug effects , Gene Expression Regulation, PlantABSTRACT
In the study, effects of S. officinalis essential oil on growth performance, health and antioxidant activity in C. carpio were investigated. The fish (13 ± 0.21 g) were fed with diet containing 1 and 3 ml kg- 1 of sage oil for 60 days. At the end of study, growth performance was not affected in fish fed with sage essential oil (p > 0.05). Superoxide dismutase (SOD) activity in hepatopancreas increased with addition of 1ml kg- 1 sage oil to the diet. However, Catalase (CAT) activity and malondialdehyde (MDA) values were not significantly altered in common carp. Total protein, albumin, glucose and hepatopancreas enzymes (aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP)) in blood serum were not affected by sage essential oil supplementation. At the histological examinations, no pathological findings were observed in hepatopancreas and intestine of carp. Goblet cells number and villi length in intestine increased with sage supplementation (p < 0.001). In addition, fertility, granulation and number of follicles increased in common carp fed with sage essential oil. Mortality after challenged with A. hydrophila was not observed in carp fed with 1ml kg- 1 concentration of sage essential oil. As a result, use of sage oil can be recommended in carp farming to improve gut health, provide disease resistance against A. hydrophila infection, and increase of fertility.
Subject(s)
Carps , Oils, Volatile , Salvia officinalis , Animals , Antioxidants/pharmacology , Dietary Supplements , Salvia officinalis/metabolism , Oils, Volatile/pharmacology , Diet/veterinary , Animal Feed/analysisABSTRACT
The present study investigated the potential ability of Salvia officinalis, one of the oldest medicinal plants, to protect male rats against cadmium reproductive toxicity. Twenty-eight healthy male rats were randomly allocated into four groups (n = 7); control, Salvia-extract treated group, cadmium treated group and a group treated with both Cd and Salvia. Administration of cadmium reduced the relative testis to body weight and significantly affected sperm parameters by decreasing motility, viability, count and increasing morphological aberrations. Comet assay was used to detect DNA fragmentation in sperms of the rats exposed to Cd. Serum levels of testosterone T, follicle stimulating hormone FSH, and luteinizing hormone LH were significantly decreased. The biochemical analysis of testicular tissue showed a significant rise in Malondialdehyde MDA level coupled with a decrease in the activity of antioxidant enzymes (superoxide dismutase SOD, glutathione peroxidase GPx and catalase CAT). The histological examination of testis sections after Cd administration revealed severe degeneration of spermatogenic cells. Seminiferous tubules were filled with homogenous eosinophilic fluid associated with atrophy of other seminiferous tubules. Co-treatment with the Salvia officinalis extract restored the oxidative enzymes activities and decreased the formation of lipid peroxidation byproduct, which in turn ameliorated the effect of Cd on sperm parameters, DNA damage and testis histopathology. Taken together, it can be concluded that the synergistic antioxidant and radical savaging activities of Salvia officinalis prevented the effect of Cd on semen quality, sperm DNA damage, along with the oxidative stress and histological abnormalities in the testis tissues.
Subject(s)
Antioxidants , Salvia officinalis , Rats , Male , Animals , Antioxidants/pharmacology , Antioxidants/metabolism , Cadmium/metabolism , Salvia officinalis/metabolism , Semen Analysis , Testis/metabolism , Spermatozoa/metabolism , Oxidative Stress , Testosterone , Sperm Motility , Seeds/metabolismABSTRACT
The diversity of plant monoterpenes is largely based on the catalytic activity of monoterpene synthases. Additionally, copy number variation of monoterpene synthase genes may contribute to the quantity of transcripts and hence to the essential oil profile. This study used whole-genome sequencing and digital PCR for the measurement of copy number variation and quantification of gene expression in three closely related Salvia species, namely Salvia officinalis, Salvia pomifera and Salvia fruticosa. Twelve, 13 and 15 monoterpene synthase-encoding open-reading frames were predicted for Salvia officinalis, Salvia pomifera and Salvia fruticosa, respectively. In Salvia officinalis, one of the open reading frames was disrupted indicating a pseudogene. Monoterpene synthase genes were generally single copy per haploid genome, only a few were double or triple copy genes. Expression levels of monoterpene synthases in leaves corresponded generally well with essential oil composition. In some cases, a higher expression level of a certain monoterpene synthase could be explained by its duplication or triplication. The very high content of thujones in Salvia pomifera, for example, was accompanied by gene duplication and increased gene expression of (+)-sabinene synthase responsible for the thujone precursor sabinene. In Salvia officinalis, three individuals different in their essential oil profile showed significant differences in their monoterpene synthase expression levels corresponding roughly to the profile of the essential oils. Transcript expression of monoterpene synthase genes were measured in leaf, calyx and corolla. The corolla differed significantly from leaves, while calyces usually showed a profile intermediary between leaf and corolla.
Subject(s)
Oils, Volatile , Salvia officinalis , Salvia , Salvia officinalis/genetics , Salvia officinalis/metabolism , DNA Copy Number Variations , Monoterpenes/metabolism , Salvia/genetics , Salvia/metabolism , Oils, Volatile/metabolismABSTRACT
The current perspective is a pioneer to assess the efficacy of Salvia officinalis leave powder (SOLP) on growth, intestinal enzymes, physiological and antioxidant status, immunological response, and gene expression of Common carp (Cyprinus carpio). We also looked into fish resistance after being challenged with Aeromonas sobria, a pathogenic zoonotic bacteria. Fish (N = 120) were fed four different experimental diets in triplicate for 8 weeks. The control diet (SOLP0 - without SOLP); meanwhile, the other three diets included SOLP of 2, 4, and 8 g kg-1 concentrations (SOLP2, SOLP4, and SOLP8), respectively. Findings demonstrated that fish fed SOLP4 and SOLP8 diets had better growth performance and improved digestion by noticeable enhancing lipase and amylase enzymes activity than other groups. Additionally, the antioxidant (superoxide dismutase and glutathione peroxidase) and immune activities (immunoglobulin M, nitric oxide, and antiprotease) clarified a significant increase (p < 0.05) in SOLP4 and SOLP8 groups. Enriched diets with SOLP4 and SOLP8 exhibited better expression of splenic genes (IL-1ß, IL-6, IL-10, TLR-2, and SOD), intestinal genes (Slc26a6) and (PepT1 or Slc15a1), and muscular genes (IGF-1 and SOD), while MSTN was down-regulated. After 8 weeks of the experimental trial, C. carpio challenged by A. sobria exhibited the highest cumulative mortality (66.67%), while SOLP8-dietary intervention showed the best results in enhancing the fish resistance against A. sobria by lessening mortalities to 13.33% followed by SOLP4 diet (20%). The outcomes indicate that the expression of splenic, muscular, and intestinal genes confirm the efficacy of SOLP on enhancing growth, digestion, and immune-antioxidant status, and recommend the potential use of SOLP especially at 4 g kg-1 level as a valuable natural economic diet additive in C. carpio culture for sustaining aquaculture.
Subject(s)
Carps , Fish Diseases , Gram-Negative Bacterial Infections , Salvia officinalis , Aeromonas , Aeromonas hydrophila/physiology , Animal Feed/analysis , Animals , Antioxidants/metabolism , Carps/metabolism , Diet/veterinary , Dietary Supplements/analysis , Gram-Negative Bacterial Infections/veterinary , Salvia officinalis/metabolism , Superoxide DismutaseABSTRACT
Vanadium has been shown to catalyze the generation of reactive oxygen species. Since free radical production and lipid peroxidation are potentially important mediators in testicular physiology and pathophysiology, the present study was conducted to elucidate vanadium-induced oxidative damage in rat testis and the ameliorative role of Salvia officinalis essential oil (SEO) against the adverse effects of this heavy metal. Adult male Wistar rats were treated daily during 10 days either with ammonium metavanadate (5 mg/kg bw, intraperitoneally), SEO (15 mg/kg bw, orally) or their combination. A group of rats receiving daily a saline solution served as a negative control. Vanadium treatment induced a significant decrease in body and reproductive organ weights, serum testosterone level and sperm number and motility. An increase in lipid peroxidation and protein oxidation as well as a marked inhibition in the activities of antioxidant enzymes in the testes and seminal vesicles indicated the occurrence of oxidative stress after vanadium toxicity. Histopathological changes in testis and seminal vesicles were also observed following vanadium administration. However, co-administration of SEO to vanadium-treated rats resulted in an appreciable improvement of these parameters, emphasizing the therapeutic effects of SEO. It can be suggested that SEO mitigates vanadium-induced reproductive damage due to its antioxidant capacity. Thus, we can hypothesize that SEO supplementation could protect against vanadium poisoning.
Subject(s)
Oils, Volatile , Salvia officinalis , Animals , Antioxidants/pharmacology , DNA Damage , Lipid Peroxidation , Male , Oils, Volatile/pharmacology , Oxidative Stress , Rats , Rats, Wistar , Reactive Oxygen Species , Saline Solution/pharmacology , Salvia officinalis/metabolism , Seeds/metabolism , Testosterone/pharmacology , Vanadium/pharmacologyABSTRACT
BACKGROUND: Cognitive health is of great interest to society, with neuroinflammation and systemic inflammation age-related risk factors that are linked to declines in cognitive performance. Several botanical ingredients have been suggested to have benefits in this area including Salvia officinalis (sage), which has shown anti-inflammatory effects and exhibited promising cognitive improvements in multiple human studies. The current study demonstrates anti-inflammatory effects for S. officinalis across a broad set of in vitro models in human cells, and adds further evidence to support modulation of acetylcholine and monoamine neurostransmitter levels as mechanisms that contribute towards the benefits of the herb on cognitive health. METHODS: The effect of S. officinalis extract on release of multiple cytokines and chemokines was measured in human primary intestinal epithelial cells treated with or without LPS stimulation, and Blood Brain Barrier (BBB) cells in presence or absence of recombinant IL-17A and/or Human IL-17RA/IL-17R Antibody. Antioxidant effects were also assessed in BBB cells incubated with the extract and H2O2. The anti-inflammatory effects of S. officinalis extract were further assessed based on clinically-relevant biomarker readouts across 12 human primary cell-based disease models of the BioMAP Diversity PLUS panel. RESULTS: S. officinalis showed significant attenuation of the release of most cytokines/chemokines into apical media in LPS-stimulated intestinal cells, but small increases in the release of markers including IL-6, IL-8 in basolateral media; where TNF-α was the only marker to be significantly reduced. S. officinalis attenuated the release of CRP and VCAM-1 from BBB cells under IL-17A induced conditions, and also decreased H2O2 induced ROS overproduction in these cells. Phenotypic profiling with the BioMAP Diversity PLUS Panel identified additional anti-inflammatory mediators, and based on a similarity search analysis suggested potential mechanistic similarity to caffeic acid and drugs known to inhibit COMT and MAO activity to modulate monoamine metabolism. Subsequent in vitro assessment showed that S. officinalis was able to inhibit the activity of these same enzymes. CONCLUSIONS: S. officinalis extract showed anti-inflammatory effects across multiple human cell lines, which could potentially reduce peripheral inflammation and support cognitive health. S. officinalis extract also showed the ability to inhibit enzymes related to the metabolism of monoamine neurotransmitters, suggesting possible dopaminergic and serotonergic effects acting alongside proposed cholinergic effects to mediate acute cognitive performance benefits previously demonstrated for the extract.
Subject(s)
Salvia officinalis , Anti-Inflammatory Agents/pharmacology , Cytokines/metabolism , Humans , Hydrogen Peroxide , Inflammation/metabolism , Interleukin-17/therapeutic use , Lipopolysaccharides/adverse effects , Plant Extracts/pharmacology , Plant Extracts/therapeutic use , Salvia officinalis/metabolismABSTRACT
The individual impact of silicon (Si) and nitric oxide (NO) on secondary metabolism in several plant species has been reported, but their combined effect has never been evaluated yet. Therefore, in this study, single and combined impacts of NO and Si on the biosynthesis of rosmarinic acid (RA) and essential oil (EO) content in leaves of Salvia officinalis were investigated under both non-stress and Cu stress conditions. The results indicated that high Cu concentration decreased biomass and the content of polyphenols, but elevated electrolyte leakage, while lower Cu concentrations, especially 200 µM Cu, increased the content of polyphenols, EO, and antioxidant capacity in leaves of S. officinalis. The foliar application of sodium silicate (1 mM Si) and sodium nitroprusside (200 µM SNP as a NO donor) alone and particularly in combination improved shoot dry biomass, restored chlorophyll and carotenoids, increased EO content, the amounts of flavonoids, and phenolic compounds especially RA, and enhanced antioxidant capacity in the leaves of S. officinalis under both non-stress and Cu stress conditions. Copper treatment increased NO content, upregulated expression of PAL, TAT, and RAS genes, and enhanced phenylalanine ammonia-lyase activity in the leaves, which were responsible for improving the production of phenolic compounds, particularly rosmarinic acid. Foliar spraying with Si and SNP intensified these attributes. All responses were more pronounced when NO and Si were simultaneously applied under Cu stress. These findings suggest that NO and Si synergistically modulate secondary metabolism through upregulation of related gene expression and enzyme activities under both non-stress and Cu stress conditions.
Subject(s)
Oils, Volatile , Salvia officinalis , Antioxidants/metabolism , Cinnamates , Depsides , Nitric Oxide/metabolism , Oils, Volatile/metabolism , Oils, Volatile/pharmacology , Polyphenols/metabolism , Salvia officinalis/genetics , Salvia officinalis/metabolism , Silicon , Rosmarinic AcidABSTRACT
To study the effects of foliar application of putrescine (distilled water (0), 0.75, 1.5, and 2.25 mM) and water deficit stress (20%, 40%, 60%, and 80% available soil water depletion (ASWD)) on the physiological, biochemical, and molecular attributes of Salvia officinalis L., a factorial experiment was performed in a completely randomized design with three replications in the growth chamber. The results of Real-Time quantitative polymerase chain reaction (qRT-PCR) analysis showed that putrescine concentration, irrigation regime, and the two-way interaction between irrigation regime and putrescine concentration significantly influenced cineole synthase (CS), sabinene synthase (SS), and bornyl diphosphate synthase (BPPS) relative expression. The highest concentration of 1,8-cineole, camphor, α-thujone, ß-thujone, CS, SS, and BPPS were obtained in the irrigation regime of 80% ASWD with the application of 0.75 mM putrescine. There was high correlation between expression levels of the main monoterpenes synthase and the concentration of main monoterpenes. The observed correlation between the two enzyme activities of ascorbate peroxidase (APX) and catalase (CAT) strongly suggests they have coordinated action. On the other hand, the highest peroxidase (PO) and superoxide dismutase (SOD) concentrations were obtained with the application of 0.75 mM putrescine under the irrigation regime of 40% ASWD. Putrescine showed a significant increase in LAI and RWC under water deficit stress. There was an increasing trend in endogenous putrescine when putrescine concentration was increased in all irrigation regimes. Overall, the results suggest that putrescine may act directly as a stress-protecting compound and reduced H2O2 to moderate the capacity of the antioxidative system, maintain the membrane stability, and increase secondary metabolites under water deficit stress.
Subject(s)
Putrescine/metabolism , Salvia officinalis/physiology , Stress, Physiological , Water , Gene Expression Regulation, Plant , Salvia officinalis/genetics , Salvia officinalis/metabolism , Terpenes/metabolismABSTRACT
<b>Background and Objective:</b> Osteoporosis is a progressive metabolic disorder characterized by an impaired bone formation that leads to increased morbidity and mortality.<i> Salvia officinalis </i>is a source of phytoestrogens that could help mitigate the risk of osteoporotic rat fracture by exerting sex hormones. Therefore, the present study was designed to investigate the curative effect of <i>Salvia officinalis </i>Extract<i> </i>(SOE) and<i> </i>17ß-estradiol (E<sub>2</sub>) and their combination<i> </i>on bone loss in female rats with ovariectomy-induced estrogen deficiency <b>Materials and Methods:</b> Forty adult female albino rats were divided into five groups, which included Sham control (Sham), ovariectomy (OVX), OVX+SOE, OVX+E<sub>2</sub> and OVX +SOE+E<sub>2</sub>.<i> </i>SOE (10 mL kg<sup></sup><sup>1</sup>) and E<sub>2</sub> (30 µg kg<sup></sup><sup>1</sup>) had been daily gavaged in the OVX+SOE, OVX+E<sub>2</sub> and OVX+SOE+E<sub>2</sub>, respectively for 6-weeks. <b>Results:</b> The model of ovariectomy resulted in osteoporosis as demonstrated by the decreased serum Ca, P, vitamin D, E<sub>2</sub> level associated with a significant increase in PTH levels in comparison to the sham control group. Besides, OVX to rats caused up-regulation in the levels of CTX-1, P1NP, BALP, OC and RANKL comparable to the sham control group. Moreover, SOE and E<sub>2</sub> significantly modulated the calciotropic parameters and improved all bone turnover markers as well as RANKL as compared to the OVX group. However, Histopathological and immunohistochemical results showed defective mineralization with the destruction of the bone matrix and increased TNF-α expression from the OVX group relative to the treated groups. <b>Conclusion:</b> These results suggest that both SOE and E<sub>2</sub> or their combined administration are efficient inhibitors against ovariectomy-induced bone loss in female rats.
Subject(s)
Estradiol/pharmacology , Osteoporosis/drug therapy , Ovariectomy/adverse effects , Plant Extracts/therapeutic use , Salvia officinalis/metabolism , Animals , Bone Density/drug effects , Disease Models, Animal , Estradiol/metabolism , Estradiol/therapeutic use , Osteoporosis/etiology , Ovariectomy/methods , Plant Extracts/metabolism , Plant Extracts/pharmacology , RatsABSTRACT
As part of our studies on antiprotozoal activity of approved herbal medicinal products, we previously found that a commercial tincture from Salvia officinalis L. (common Sage, Lamiaceae) possesses high activity against Trypanosoma brucei rhodesiense (Tbr), causative agent of East African Human Trypanosomiasis. We have now investigated in detail the antitrypanosomal constituents of this preparation. A variety of fractions were tested for antitrypanosomal activity and analyzed by UHPLC/+ESI QqTOF MS. The resulting data were used to generate a partial least squares (PLS) regression model that highlighted eight particular constituents that were likely to account for the major part of the bioactivity. These compounds were then purified and identified and their activity against the pathogen tested. All identified compounds (one flavonoid and eight diterpenes) displayed significant activity against Tbr, in some cases higher than that of the total tincture. From the overall results, it can be concluded that the antitrypanosomal activity of S. officinalis L. is, for the major part, caused by abietane-type diterpenes of the rosmanol/rosmaquinone group.
Subject(s)
Antiprotozoal Agents/pharmacology , Salvia officinalis/metabolism , Trypanosoma brucei rhodesiense/drug effects , Abietanes/chemistry , Animals , Calibration , Chromatography, High Pressure Liquid , Diterpenes/chemistry , Flavonoids/chemistry , Inhibitory Concentration 50 , Least-Squares Analysis , Magnetic Resonance Spectroscopy , Muscle, Skeletal/metabolism , Myoblasts/metabolism , Parasitic Sensitivity Tests , Plant Extracts , Plant Preparations/pharmacology , Plants, Medicinal , Rats , Spectrometry, Mass, Electrospray Ionization , Trypanosomiasis/parasitologyABSTRACT
Being aware of the remarkable antimicrobial potential of S. officinalis L., we aimed to evaluate the antimicrobial activity of the S. officinalis dichloromethane crude extract (SOD), dichloromethane-soluble fractions (SODH and SODD), SODD subfractions (SODD1 and SODD2), and pure substances (manool, salvigenin, and viridiflorol) against periodontopathogens. This bioassay-guided study comprises five antimicrobial tests-determination of the Minimum Inhibitory Concentration (MIC), determination of the Minimum Bactericidal Concentration (MBC), determination of the antibiofilm activity, construction of the Time-kill curve (determination of Bactericidal Kinetics), and determination of the Fractional Inhibitory Concentration Index-on six clinical bacterial isolates and three standard bacterial strains involved in periodontal disease. SOD has moderate activity against most of the tested bacteria, whereas SODD1, SODH1, SODH3, and manool afford the lowest results. The Porphyromonas gingivalis (ATTC and clinical isolate) biofilm is considerably resistant to all the samples. In association with chlorhexidine gluconate, only SODH1 exerts additive action against P. gingivalis (clinical isolate). Therefore, SODH1 and manool are promising antibacterial agents and may provide therapeutic solutions for periodontal infections.
Subject(s)
Aggressive Periodontitis , Anti-Bacterial Agents/pharmacology , Plant Extracts/pharmacology , Salvia officinalis/metabolism , Aggressive Periodontitis/drug therapy , Aggressive Periodontitis/microbiology , Bacteria/drug effects , Biofilms/drug effects , Diterpenes/pharmacology , Humans , Microbial Sensitivity Tests , Mouth/microbiology , Porphyromonas gingivalis/drug effectsABSTRACT
We aimed in the present study to investigate the chemical composition, the antioxidant capacities as well as the inâ vitro fermentation properties of Salvia officinalis leaves aqueous extract (SOLAE) grown in four regions of northwestern Tunisia. Our data firstly indicated a spatial variation (P<0.05) in condensed tannins, total lipids, polyphenols and flavonoids contents. The HPLC-PDA-ESI-MS/MS-LC/HR-ESI-MS technique allowed to the identification of 13 phenolic compounds and showed that protocatechuic acid is the major constituent of the plant leaves grown in Tabarka, Ain Draham and Testour. The SOLAE of the plant grown in Tabarka presents the most potent scavenging activity against DPPH radical and had the highest percentage of inhibition. More importantly, we found in the present study that the digestibility of dry matter and inâ vitro fermentation showed a significant variation between the regions and the animal species. Also, we showed a very positive correlation between antioxidant properties and phenolic compounds contents. In conclusion, we suggest that SOLAE had potential beneficial effects owing in part to its antioxidant and ROS scavenging activities. Therefore, S. officinalis can be proposed as an additive food for animals' nutrition and health.
Subject(s)
Antioxidants/pharmacology , Biphenyl Compounds/antagonists & inhibitors , Fermentation , Phytochemicals/pharmacology , Picrates/antagonists & inhibitors , Salvia officinalis/metabolism , Animals , Antioxidants/analysis , Antioxidants/metabolism , Biphenyl Compounds/metabolism , Chromatography, High Pressure Liquid , Drug Evaluation, Preclinical , Goats , Hydroxybenzoates/analysis , Hydroxybenzoates/metabolism , Hydroxybenzoates/pharmacology , Mass Spectrometry , Phenol/analysis , Phenol/metabolism , Phenol/pharmacology , Phytochemicals/analysis , Phytochemicals/metabolism , Picrates/metabolism , Plant Leaves/chemistry , Plant Leaves/metabolism , Salvia officinalis/chemistry , Salvia officinalis/growth & development , Sheep , TunisiaABSTRACT
In the present study, aqueous extracts of Salvia africana, Salvia officinalis 'Icterina' and Savia mexicana origin were screened for their phenolic composition and for antibacterial, antioxidant, anti-inflammatory and cytotoxic properties. The three aqueous extracts contained distinct phenolic compounds, with S. africana presenting the highest total levels (231.6 ± 7.5 µg/mg). Rosmarinic acid was the dominant phenolic compound in all extracts, yet that of S. africana origin was characterized by the present of yunnaneic acid isomers, which overall accounted for about 40% of total phenolics. In turn, S. officinalis 'Icterina' extract presented glycosidic forms of apigenin, luteolin and scuttelarein, and the one obtained from S. mexicana contained several simple caffeic acid derivatives. S. africana aqueous extract exhibited high antioxidant potential in four methods, namely the DPPH⢠(2,2-diphenyl-1-picrylhydrazyl) scavenging ability, iron-reducing power, inhibition of ß-carotene bleaching and of thiobarbituric acid reactive substances (TBARS), for which EC50 values were equal or only 1.3-3.1 higher than those of the standard compounds. Moreover, this extract was able to lower the levels of nitric oxide (NO) production in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages (EC50 = 47.8 ± 2.1 µg/mL). In addition, the three sage aqueous extracts showed promising cytotoxic effect towards hepatocellular HepG2, cervical HeLa, and breast carcinoma cells MCF-7. Overall this study highlights the potential of three little-exploited Salvia species, with commercial value for applications in food or pharmaceutical industries.
Subject(s)
Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/pharmacology , Phenols/analysis , Salvia officinalis/chemistry , Anti-Bacterial Agents/pharmacology , Anti-Inflammatory Agents/pharmacology , Antioxidants/chemistry , Biphenyl Compounds/chemistry , Biphenyl Compounds/pharmacology , Camphanes , Cinnamates/chemistry , Cytotoxins/pharmacology , Depsides/chemistry , Free Radical Scavengers/pharmacology , HeLa Cells , Hep G2 Cells , Humans , MCF-7 Cells , Panax notoginseng , Phenols/chemistry , Phytochemicals , Picrates/chemistry , Picrates/pharmacology , Salvia miltiorrhiza , Salvia officinalis/metabolism , Rosmarinic AcidABSTRACT
Rosemary and sage species from Lamiaceae contain high amounts of structurally related but diverse abietane diterpenes. A number of substances from this compound family have potential pharmacological activities and are used in the food and cosmetic industry. This has raised interest in their biosynthesis. Investigations in Rosmarinus officinalis and some sage species have uncovered two main groups of cytochrome P450 oxygenases that are involved in the oxidation of the precursor abietatriene. CYP76AHs produce ferruginol and 11-hydroxyferruginol, while CYP76AKs catalyze oxidations at the C20 position. Using a modular Golden-Gate-compatible assembly system for yeast expression, these enzymes were systematically tested either alone or in combination. A total of 14 abietane diterpenes could be detected, 8 of which have not been reported thus far. We demonstrate here that yeast is a valid system for engineering and reconstituting the abietane diterpene network, allowing for the discovery of novel compounds with potential bioactivity.
Subject(s)
Abietanes/metabolism , Cytochrome P-450 Enzyme System/metabolism , Rosmarinus/metabolism , Saccharomyces cerevisiae/metabolism , Salvia officinalis/chemistry , Salvia officinalis/metabolism , Abietanes/chemistry , Cytochrome P-450 Enzyme System/genetics , Molecular Structure , Oxidation-Reduction , Rosmarinus/chemistry , Saccharomyces cerevisiae/genetics , Salvia officinalis/geneticsABSTRACT
Taxadiene synthase (TXS) catalyzes the formation of natural product taxa-4(5),11(12)-diene (henceforth taxadiene). Taxadiene is the precursor in the formation of Taxol, which is an important natural anticancer agent. In the current study, we present a detailed mechanistic view of the biosynthesis of taxadiene by TXS using a hybrid quantum mechanics-molecular mechanics potential in conjunction with free energy simulation methods. The obtained free-energy landscape displays initial endergonic steps followed by a stepwise downhill profile, which is an emerging free-energy fingerprint for type I terpene synthases. We identify an active-site Trp residue (W753) as a key feature of the TXS active-site architecture and propose that this residue stabilized intermediate cations via π-cation interactions. To validate our proposed active TXS model, we examine a previously reported W753H mutation, which leads to the exclusive formation of side product cembrene A. The simulations of the W753H mutant show that, in the mutant structure, the His side chain is in the perfect position to deprotonate the cembrenyl cation en route to cembrene formation and that this abortive deprotonation is an energetically facile process. On the basis of the current model, we propose that an analogous mutation of Y841 to His could possibly lead to verticillane. The current simulations stress the importance of the precise positioning of key active-site residues in stabilizing intermediate carbocations. In view of the great pharmaceutical importance of taxadiene, a detailed understanding of the TXS mechanism can provide important clues toward a synthetic strategy for Taxol manufacturing.
Subject(s)
Alkenes/metabolism , Diterpenes/metabolism , Isomerases/metabolism , Plant Proteins/metabolism , Salvia officinalis/metabolism , Biosynthetic Pathways , Catalytic Domain , Isomerases/chemistry , Isomerases/genetics , Models, Molecular , Plant Proteins/chemistry , Plant Proteins/genetics , Point Mutation , Protein Conformation , Salvia officinalis/chemistry , Salvia officinalis/enzymology , Salvia officinalis/genetics , ThermodynamicsABSTRACT
A large number of terpenoid compounds have been extracted from different tissues of S. officinalis. However, the molecular genetic basis of terpene biosynthesis pathways is virtually unknown. In this study, approximately 6.6 Gb of raw data were generated from the transcriptome of S. officinalis leaves using Illumina HiSeq 2000 sequencing. After filtering and removing the adapter sequences from the raw data, the number of reads reached 21 million, comprising 98 million of high-quality nucleotide bases. 48,671 unigenes were assembled de novo and annotated for establishing a valid database for studying terpenoid biosynthesis. We identified 135 unigenes that are putatively involved in terpenoid metabolism, including 70 mevalonate and methyl-erythritol phosphate pathways, terpenoid backbone biosynthesis genes, and 65 terpene synthase genes. Moreover, five terpene synthase genes were studied for their functions in terpenoid biosynthesis by using transgenic tobacco; most transgenic tobacco plants expressing these terpene synthetic genes produced increased amounts of terpenoids compared with wild-type control. The combined data analyses from the transcriptome and metabolome provide new insights into our understanding of the complex metabolic genes in terpenoid-rich sage, and our study paves the way for the future metabolic engineering of the biosynthesis of useful terpene compounds in S. officinalis.
Subject(s)
Biosynthetic Pathways/genetics , Genes, Plant , Metabolomics , Salvia officinalis/genetics , Salvia officinalis/metabolism , Terpenes/metabolism , Transcriptome/genetics , Alkyl and Aryl Transferases/genetics , Alkyl and Aryl Transferases/metabolism , Gene Expression Regulation, Plant , Microsatellite Repeats/genetics , Molecular Sequence Annotation , Oils, Volatile/analysis , Phylogeny , Plant Leaves/genetics , Plants, Genetically Modified , RNA, Messenger/genetics , RNA, Messenger/metabolism , Reproducibility of Results , Nicotiana/genetics , VolatilizationABSTRACT
Composition of tocopherols, tocotrienols, carotenoids, fatty acids, as well as hydrophilic and lipophilic antioxidant activities, were determined in seeds of two Salvia species and oils obtained from them. Both seeds contained a large amount of oil (around 20%) rich in polyunsaturated fatty acids. While Salvia officinalis seed oil can be classified as oleic-linoleic oil, the predominant fatty acid in Salvia sclarea was α-linolenic acid (around 54%). Among tocols, the main isomers in both seeds and oils were γ-tocopherol, followed by α-tocopherol. Concerning carotenoids, their concentration was around 0.75 mg/100 g of seeds and 0.16 mg/100 g of oils, with a predominance of lutein. Oil and seeds of S. officinalis exhibited higher antioxidant potential compared to S. sclarea investigated samples which could be attributed to higher content of total vitamin E and carotenoids. This study provides results that enables use of two Salvia species as new alternative sources of vegetable oils.
Subject(s)
Antioxidants/chemistry , Oils, Volatile/chemistry , Salvia/metabolism , Antioxidants/analysis , Carotenoids/analysis , Chromatography, Gas , Chromatography, High Pressure Liquid , Gas Chromatography-Mass Spectrometry , Oils, Volatile/analysis , Salvia/chemistry , Salvia officinalis/chemistry , Salvia officinalis/metabolism , Seeds/chemistry , Seeds/metabolism , alpha-Tocopherol/analysisABSTRACT
Four essential oils (EOs) from Salvia officinalis L. cultivated in Spain (Murcia Province) were analyzed by gas chromatography coupled with mass spectrometry (GC/MS) to determine their relative and absolute compositions. The main components were α-thujone (22.8 - 41.7%), camphor (10.7 - 19.8%), 1,8-cineole (4.7 - 15.6%), and ß-thujone (6.1 - 15.6%). Enantioselective gas chromatography identified (-)-α-thujone and (+)-camphor as the main enantiomers in all the analyzed EOs. Furthermore, when the EOs were tested to determine their antioxidant activity against free radicals and as ferric reducing and ferrous chelating agents, all were seen to have moderate activity due to the compounds they contained, such as linalool or terpinene. Because of their known relation with inflammatory illnesses and Alzheimer's disease, respectively, the inhibition of lipoxygenase and acetylcholinesterase was studied using the EOs. Some individual compounds also inhibited these enzymes. In addition, the studied EOs were able to inhibit the growth of Escherichia coli, Staphylococcus aureus, and Candida albicans. The characterization carried out increases our awareness of the possible uses of S. officinalis EO as natural additives in food, cosmetics, and pharmaceuticals.
Subject(s)
Oils, Volatile/chemistry , Salvia officinalis/chemistry , Acetylcholinesterase/chemistry , Acetylcholinesterase/metabolism , Acyclic Monoterpenes , Anti-Infective Agents/chemistry , Anti-Infective Agents/isolation & purification , Anti-Infective Agents/pharmacology , Antioxidants/chemistry , Bicyclic Monoterpenes , Candida albicans/drug effects , Cluster Analysis , Cyclohexanols/analysis , Escherichia coli/drug effects , Eucalyptol , Gas Chromatography-Mass Spectrometry , Inhibitory Concentration 50 , Lipoxygenase/chemistry , Lipoxygenase/metabolism , Monoterpenes/analysis , Oils, Volatile/analysis , Oils, Volatile/metabolism , Principal Component Analysis , Protein Binding , Salvia officinalis/metabolism , Spain , Staphylococcus aureus/drug effects , StereoisomerismABSTRACT
MAIN CONCLUSION: As a result of this work, we were able to characterize seven indigenous to Greece Salvia officinalis populations using genetic and metabolomic tools. These tools can be used to select the most promising genotypes, capable to design future breeding programs for high valuable varieties. An initial investigation was carried out to compare the genetic and metabolic diversity in S. officinalis grown in Greece and to discern the relationship between the two sets of data. Analysis of inter-simple sequence repeats (ISSR) revealed significant genetic differences among seven sage populations, which were grouped into three main clusters according to an UPGMA ISSR data-based dendrogram and Principle Coordinate Analysis. 80 loci were scored of which up to 90% were polymorphic at species level. According to the composition of their essential oil, the populations were classified into two chemotypes: 1.8 cineole/α-thujone and α-thujone/1.8 cineole. Additionally, a targeted ultra performance liquid chromatography (UPLC-MS/MS) method was used to qualify and quantify phenolic compounds in methanolic extracts of the seven sage genotypes according to which they were districted in six clusters among the sage populations. The main compounds characterizing the seven genotypes were rosmarinic acid and carnosol, followed by apigenin-7-O-glucoside (Ap7glc), and luteolin-7-O-glucoside (Lu7glc). The correlation between matrices obtained from ISSR data and metabolic profiles was non-significant. However, based on the differences in metabolic fingerprint, we aimed to define populations using as main selection criteria the high polyphenol content and desired essential oil composition, using state to the art analytical tools for the identification of parent lines for breeding programs.