Inhibition Of Nitric Oxide Release Research Articles

Study on the Chemical Components and Anti-neuroinflammatory Activity of the Roots of Clausena excavate Burm. f.

The ethanol extract of the roots of Clausena excavata gave two previously undescribed coumarins, clauexcatin A (1) and clauexcatin B (2), as well as a pair of new isomers, trans/cis-clauexcatin C (3a, 3b), along with thirty known compounds. Among these, compound 33 was isolated from this genus for the first time. The structures of these compounds were elucidated based on their physicochemical properties and spectroscopic data. The anti-neuroinflammatory activities were assessed using LPS-activated BV-2 microglial cells. Compounds 6, 8, 17, 24, 29, and 30 exhibited significant inhibition of nitric oxide release in a dose-dependent manner, with their inhibitory effects being 1.2 to 10.9 times greater than that of the positive control (minocycline).

Chemical composition and the anti-inflammatory effect of volatile compounds from Anaxagorea luzonensis A. Gray.

Anaxagorea luzonensis A. Gray, a member of the Annonaceae family, has been used to treat a variety of illnesses for a long time. For the first time, A. luzonensis volatile compounds (ALVCs) were extracted from the leaves,and the components were identified using gas chromatography-mass spectrometry (GC-MS). Further, the main compositions of ALVCs were also assessed for their ability to bind with anti-inflammatory proteins using a docking model. In addition, invitro tests e.g. inhibition of protein degradation and the inhibition of nitric oxide release using RAW264.7 macrophage cells were utilized for evaluating the anti-inflammatory activity. The results showed that the principal compounds of ALVCs were bulnesol (34.1 %), cubitene (17.8 %), β-eudesmol (10.4 %), epi-longipinanol (5.9 %), and (Z)-nerolidyl acetate (5.5 %). Three compounds viz. bulnesol, cubitene, and β-eudesmol bound firmly to cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX), as shown by the in silico analysis, similar to the positive control diclofenac. ALVCs effectively inhibited protein degradation with the IC50 of 31±2.3 μg/mL and inhibited nitric oxide production with the IC50 of 43.30±3.37 μg/mL. These findings showed that ALVCs might have a promising anti-inflammatory effect by blocking several inflammatory proteins.

Exploring anti-inflammatory and antioxidant-related quality markers of Artemisia absinthium L. based on spectrum-effect relationship.

Artemisia absinthium L. is a well-known medicinal, aromatic, and edible plant with important medicinal and economic properties and a long history of use in treating liver inflammation and other diseases; however, there has been insufficient progress in quality control. This study aimed to investigate the quality markers for the anti-inflammatory and antioxidant activities of A.absinthium based on spectrum-effect relationship analysis. Eighteen batches of A.absinthium from different origins were used. Chemical fingerprints were obtained by ultra-performance liquid chromatography (UPLC). The chemical compositions were identified by quadrupole-Orbitrap high-resolution mass spectrometry. Anti-inflammatory activity was assessed by inhibition of cyclooxygenase-2 and 15-lipoxygenase in vitro and inhibition of nitric oxide release in lipopolysaccharide-induced BV-2 cells. Antioxidant activity was assessed by DPPH and ABTS radical scavenging assays. The relationship between bioactivity and chemical fingerprints was then analyzed using chemometrics including gray relational analysis, bivariate correlation analysis, and orthogonal partial least squares analysis. Different batches of A.absinthium extracts possessed significant anti-inflammatory and antioxidant activities to varying degrees. Eighty compounds were identified from A.absinthium, and 12 main common peaks were obtained from the UPLC fingerprints. P3 (chlorogenic acid), P5 (isochlorogenic acid A), and P6 (isochlorogenic acid C) were screened as the most promising active compounds by correlation analysis and further validated for their remarkable anti-inflammatory effects. This is the first study to screen the quality markers of A.absinthium by establishing the spectrum-effect relationship, which can provide a reference for the development of quality standards and further research on A.absinthium.

Anti-inflammatory activity of seven plant species with potential use as livestock feed additives

Inflammatory conditions and associated oxidative stress are common in livestock. The present study aimed to investigate in depth the anti-inflammatory activity of seven medicinal plants (Dichrostachys cinerea, Ehretia rigida, Salix babylonica, Vachellia erioloba, V. gerrardii, V. sieberiana and V. tortilis) traditionally used to treat inflammatory disorders in South Africa. These plants are being investigated for possible inclusion in animal feed as additives to support health and promote growth. The anti-inflammatory potential of plant extracts was evaluated in terms of inhibition of the 15-lipoxygenase (15-LOX) enzyme and nitric oxide (NO) release in LPS-induced RAW 264.7 macrophages, together with cytotoxicity studies. The effect of plant extracts in modulating the expression of pro-inflammatory mediators including cyclooxygenase (COX), inducible nitric oxide synthase (iNOS), and the cytokine interleukin-6 (IL-6) was evaluated using quantitative reverse transcription polymerase chain reaction (qRT-PCR). In vitro antioxidant activity was investigated using chemical assays. ULPC-MS and ICP-MS were used to detect potentially bioactive phytochemicals and concentrations of essential elements respectively. Methanol and acetone extracts of D. cinerea had the best activity against 15-LOX (IC50 values of 0.80 and 0.64 mg/ml, respectively). The methanol extracts of E. rigida, V. tortilis and V. sieberiana inhibited NO with IC50 of 90.11, 101.52 and 94.11 µg/ml respectively, higher than that of the positive control (IC50 = 30.00 µg/ml). Furthermore, these plants had low cytotoxicity (LC50 ≥ 70 µg/ml) against RAW 264.7 macrophages. Interestingly, E. rigida, V. sieberiana and V. tortilis extracts decreased the expression of iNOS, COX and IL-6. There was a positive correlation between the inhibition of NO release from macrophages by plant extracts and the downregulation of iNOS mRNA levels. There was no significant difference in the antioxidant activity (ABTS) of D. cinerea methanolic extract with IC50 = 5.37 µg/ml compared to the positive controls, Trolox (IC50 = 4.42 µg/ml) and ascorbic acid (IC50 = 4.43 µg/ml). Flavonoids, fatty acids and essential trace elements such as Fe, Zn and Mn were detected in E. rigida, and these are likely to be responsible for the good anti-inflammatory effect of the extracts. This study provides the first evidence of the regulation of pro-inflammatory mediator and cytokine genes by the selected plant extracts. These results support the use of the plants, in particular E. rigida, as potential feed additives to reduce inflammation associated with many diseases.

Enzymatic upcycling of wild-simulated ginseng leaves for enhancing biological activities and compound K

Compound K (CK), a ginsenoside with high bioavailability, is present at low levels in wild-simulated ginseng leaves (WSGL). WSGL contains the CK precursors, Rd and F2, in amounts up to 26.4 ± 0.4 and 24.1 ± 1.9 mg/g extract, respectively. In this study, CK production in WGSL reached 25.9 ± 1.0 mg/g extract following treatment with Viscozyme, Celluclast 1.5 L, Pectinex Ultra SP-L, and their combination. The antioxidant activities indicated by oxygen radical absorbance capacity, ferric reducing antioxidant power, and ABTS- and DPPH radical scavenging activity of enzyme-treated WSGL were enhanced 1.69-, 2.51-, 2.88-, and 1.80-fold, respectively, compared to non-treated WSGL. Furthermore, the CK-enriched WSGL demonstrated a 1.94-fold decrease in SA-β-galactosidase expression in human dermal fibroblasts and a 3.8-fold enhancement of inhibition of nitric oxide release in lipopolysaccharide-induced RAW 264.7 cells relative to non-treated WSGL. Consequently, WSGL subjected to enzymatic upcycling has potential as a functional material in the food and pharmaceutical industries.

The Effect of Ozone Dosage of Sunflower and Olive Oils on Their Biological Activities and Chemical Properties

ABSTRACT The ozonation process of vegetable oils is commonly monitored by peroxide value (PV). However, PV methodology has been adapted from vegetable oils, with protocol divergences. In addition, few studies are based on the minimum level of ozonation to ensure biological activity. This study aimed to optimize the PV methodology for sunflower oil with different ozone dosage aiming to discriminate these samples. Also, to establish the relationship between its physicochemical properties with the biological activity. The optimized PV method was also applied to ozonated olive oil, as well as the other analytical techniques of iodine value (IV), acidity value (AV), density, viscosity, gas chromatography, infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR), differential scanning calorimetry and thermogravimetry (DSC and TG). The PV method discriminated the ozonation level of the samples. Both ozonated sunflower and olive oils need to reach an intermediary ozone dosage to perform the in vitro bactericidal and fungicidal activity (MIC ≤2.5%), and anti-inflammatory activity (inhibition of nitric oxide release in Raw cells). These results point to the importance of standardizing the ozone dosage for each oil and the analytical methodologies used in its characterization.

Up-regulated Expression of a TOR2A Gene Product-Salusin-β in the Paraventricular Nucleus Enhances Sympathetic Activity and Cardiac Sympathetic Afferent Reflex in Rats with Chronic Heart Failure Induced by Coronary Artery Ligation.

Enhanced cardiac sympathetic afferent reflex (CSAR) promotes sympathetic hyperactivation in chronic heart failure (CHF). Salusin-β is a torsin family 2 member A gene product and a cardiovascular active peptide closely associated with cardiovascular diseases. We aimed to determine the roles of salusin-β in the paraventricular nucleus (PVN) in modulating enhanced CSAR and sympathetic hyperactivation in rats with CHF induced by coronary artery ligation and elucidate the underlying molecular mechanisms. CSAR was evaluated based on the responses of mean arterial pressure (MAP) and renal sympathetic nerve activity (RSNA) to the epicardial administration of capsaicin in rats under anaesthesia. Salusin-β protein expression was up-regulated in the PVN of the CHF compared with sham-operated rats. Salusin-β microinjection into the PVN dose-dependently increased MAP and RSNA and enhanced CSAR, while anti-salusin-β IgG exerted opposite effects. The effect of salusin-β was inhibited by reactive oxygen species (ROS) scavenger or NAD(P)H oxidase inhibitor but promoted by superoxide dismutase inhibitor. The effect of anti-salusin-β IgG was interdicted by nitric oxide (NO) synthase inhibitor. Furthermore, chronic salusin-β gene knockdown in PVN attenuated CSAR, reduced sympathetic output, improved myocardial remodelling and cardiac function, decreased NAD(P)H oxidase activity and ROS levels, and increased NO levels in the CHF rats. Increased salusin-β activity in the PVN contributes to sympathetic hyperactivation and CSAR in CHF by inhibiting NO release and stimulating NAD(P)H oxidase-ROS production. Reducing endogenous central salusin-β expression might be a novel strategy for preventing and treating CHF in the future.

Chemical constituents from leaves of Jatropha curcas

ObjectiveTo investigate the chemical constituents from the leaves of Jatropha curcas and evaluate their inhibition on lipopolysaccharide (LPS)-activated BV-2 microglia cells. MethodsThe n-BuOH extract of the leaves of J. curcas was isolated by macroporous adsorption resin, silica gel, ODS, column chromatography and semi-preparative HPLC. The structures of the compounds were identified by MS, NMR, ECD, and other spectroscopic methods. In addition, anti-neuroinflammatory effects of isolated compounds were evaluated by measuring the production of nitric oxide (NO) in over-activated BV-2 cells. ResultsSeventeen compounds, including (7R,8S)-crataegifin A-4-O-β-D-glucopyranoside (1), (8R,8′R)-arctigenin (2), arctigenin-4′-O-β-D-glucopyranoside (3), (-)-syringaresinol (4), syringaresinol-4′-O-β-D-glucopyranoside (5), (-)-pinoresinol (6), pinoresinol-4′-O-β-D-glucopyranoside (7), buddlenol D (8), (2R,3R)-dihydroquercetin (9), (2S,3S)-epicatechin (10), (2R,3S)-catechin (11), isovitexin (12), naringenin-7-O-β-D-glucopyranoside (13), chamaejasmin (14), neochamaejasmin B (15), isoneochamaejasmin A (16), and tomentin-5-O-β-D-glucopyranoside (17) were isolated and identified. Compounds 2, 4 and 8 significantly inhibited the release of NO in BV-2 microglia activated by LPS, with IC50 values of 18.34, 29.33 and 26.30 μmol/L, respectively. ConclusionCompound 1 is a novel compound, and compounds 2, 3, 8, 14–17 are isolated from Jatropha genus for the first time. In addition, the lignans significantly inhibited NO release and the inhibitory activity was decreased after glycosylation.

Role of Dietary Polyphenols in the Activity and Expression of Nitric Oxide Synthases: A Review.

Nitric oxide (NO) plays several key roles in the functionality of an organism, and it is usually released in numerous organs and tissues. There are mainly three isoforms of the enzyme that produce NO starting from the metabolism of arginine, namely endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS), and neuronal nitric oxide synthase (nNOS). The expression and activity of these isoforms depends on the activation/deactivation of different signaling pathways at an intracellular level following different physiological and pathological stimuli. Compounds of natural origin such as polyphenols, which are obtainable through diet, have been widely studied in recent years in in vivo and in vitro investigations for their ability to induce or inhibit NO release, depending on the tissue. In this review, we aim to disclose the scientific evidence relating to the activity of the main dietary polyphenols in the modulation of the intracellular pathways involved in the expression and/or functionality of the NOS isoforms.

Biological Activity of Ganoderma Species (Agaricomycetes) from Sonoran Desert, Mexico.

Ganoderma species have been used in folk medicine against different illnesses and are characterized by producing a diversity of bioactive metabolites (triterpenoids, polysaccharides, flavonoids, and phenols) with numerous medicinal effects (anti-proliferative, antioxidant, anti-inflammatory, and antibacterial). This work aims to evaluate ethanolic extracts of fruiting bodies of Ganoderma oerstedii, G. weberianum, and G. subincrustatum strains from the Sonoran Desert in the anti-proliferative activity by the MTT assay on cancer cell lines; anti-inflammatory effect by quantifying nitric oxide (NO) production; antioxidant activity by DPPH, ABTS, and FRAP assays; total phenolic and flavonoid content by Folin-Ciocalteu and AlCl3 method, respectively; antibacterial activity by the broth microdilution method against Escherichia coli and Staphylococcus aureus. Extracts showed anti-proliferative activity with IC50 < 100 µg/mL on the cancer cell lines MDA-MB-231, A549, and HeLa, except G. subincrustatum extract with an IC50 > 100 µg/mL; anti-proliferative activity was not selective, being affected non-cancerous cell line ARPE-19. Extracts showed significant inhibition of NO release in cells stimulated by LPS, up to 60% with G. subincrustatum and G. oerstedii, and 47% with G. weberianum. All tested assays showed moderate antioxidant potential; the most active was G. lucium(control strain) with IC50 of 69 and 30 µg/mL by DPPH and ABTS respectively; and 271 µg Trolox equivalents/g by FRAP. Total phenols and flavonoids ranged from 38 to 56 mg GAE/g and 0.53 to 0.93 mg QE/g, respectively. A significant correlation was found between the antioxidant activities revealed by DPPH, ABTS, and FRAP with total phenol and flavonoid contents. Antibacterial activity was weak against S. aureus (MIC50 > 10 mg/mL). These results demonstrate that tested Ganoderma mushrooms have medicinal potential such as anti-inflammatory and anti-proliferative.

Elderberry (Sambucus nigra L.) extracts promote anti-inflammatory and cellular antioxidant activity.

Despite the high value of Portuguese elderberries, recognized for decades by European markets, only a few studies address their beneficial effects at cellular level. Aiming to explore the anti-inflammatory and the cellular antioxidant potential characterized extracts from the three main Portuguese elderberry cultivars (Sabugueiro, Sabugueira, Bastardeira) were used. Lipopolysaccharide-stimulated RAW 264.7 cells pre-exposed to elderberry extracts exhibited dose-dependent inhibition of nitric oxide release, evidencing anti-inflammatory activity. Concerning cellular antioxidant protection, HepG2 and Caco-2 cells pre-exposure to elderberry extracts (50µg/mL) prevented up-to 90% of tert-butyl hydroperoxide (t-BOOH)-induced toxicity. In Caco-2 cells, elderberry extracts prevented glutathione depletion, reactive oxygen species production, abnormal morphological changes and DNA fragmentation, in response to t-BOOH oxidative insult. Results demonstrated that elderberries have high potential in reducing cellular oxidative stress as well as in preventing inflammatory processes. Thus, elderberries have high potential as health promoters, acting as functional foods or as sources of nutraceuticals.

Design and synthesis of novel aza-ursolic acid derivatives: in vitro cytotoxicity and nitric oxide release inhibitory activity.

Aim: Inducible nitric oxide synthase (iNOS) is a validatedtargetfor anti-inflammatory treatment. Based on the authors' previous work,novel aza-ursolic acid derivatives were designedand synthesizedandtheir inhibitory activities against lipopolysaccharide (LPS)-induced nitric oxide (NO) release from RAW264.7 cells wasevaluated. Materials & results: 16 novel derivatives were screened for their in vitro inhibitory activity against NO release using Griess assaysand the cytotoxicity was evaluated using MTT assays. The presence of furoxan joined to theA-ring of ursolic acid and N-methylpiperazine groups in the lead compound was identified for anti-inflammatory activity, and compound21b showed 94.96% inhibition of NO release at 100μM with an IC50 value of 8.58μM. Conclusion: Compound 21b has potential anti-inflammatory activity with low cytotoxicity that warrants further preclinical study and evaluation.

Nitric oxide regulates adhesiveness, invasiveness, and migration of anoikis-resistant endothelial cells.

Anoikis is a type of apoptosis that occurs in response to the loss of adhesion to the extracellular matrix (ECM). Anoikis resistance is a critical mechanism in cancer and contributes to tumor metastasis. Nitric oxide (NO) is frequently upregulated in the tumor area and is considered an important player in cancer metastasis. The aim of this study was to evaluate the effect of NO on adhesiveness, invasiveness, and migration of anoikis-resistant endothelial cells. Here, we report that anoikis-resistant endothelial cells overexpress endothelial nitric oxide synthase. The inhibition of NO release in anoikis-resistant endothelial cells was able to decrease adhesiveness to fibronectin, laminin, and collagen IV. This was accompanied by an increase in cell invasiveness and migration. Furthermore, anoikis-resistant cell lines displayed a decrease in fibronectin and collagen IV protein expression after L-NAME treatment. These alterations in adhesiveness and invasiveness were the consequence of MMP-2 up-regulation observed after NO release inhibition. The decrease in NO levels was able to down-regulate the activating transcription factor 3 (ATF3) protein expression. ATF3 represses MMP-2 gene expression by antagonizing p53-dependent trans-activation of the MMP-2 promoter. We speculate that the increased release of NO by anoikis-resistant endothelial cells acted as a response to restrict the MMP-2 action, interfering in MMP-2 gene expression via ATF3 regulation. The up-regulation of nitric oxide by anoikis-resistant endothelial cells is an important response to restrict tumorigenic behavior. Without this mechanism, invasiveness and migration potential would be even higher, as shown after L-NAME treatment.

Anticancer profile and anti-inflammatory effect of new N-(2-((4-(1,3-diphenyl-1H-pyrazol-4-yl)pyridine sulfonamide derivatives

A new series of N-(2-((4-(1,3-diphenyl-1H-pyrazol-4-yl)pyridine sulfonamide derivatives 11a-o were designed and synthesized based on our previous works. The new series was tested for its anticancer and anti-inflammatory effects. The anticancer profile of final target compounds was obtained by testing them over 60 cell lines belong to nine types of cancers. Compound 11c showed the highest percent inhibition, so its potency was measured over the most sensitive cell line to determine its IC50 over each cell. In addition, compound 11c was tested over kinase panel to get its biological target(s). Compound 11c had strong activity over JNK1, JNK2, p38a and V600EBRAF. All final target compounds were tested against the four kinases to build a structure activity relationship. Compound 11c was subjected to cell cycle analysis to check at which phase is affected by 11c. The anti-inflammatory effect of final target compounds was screened by testing their ability to inhibit both nitric oxide release and prostaglandin E2 production on raw 264.7 macrophages in addition to test their cytotoxic effect on the same cells. Compound 11n showed the highest ability to inhibit prostaglandin E2 and all compound showed moderate to low activity regarding inhibition of nitric oxide release. Compound 11n was investigated for its ability to reduce Interleukin 6 and TNF-alpha. In addition, compound 11n was tested for its effect on induced Nitric oxide synthase (iNOS), and COX-2 mRNA expression level and its effect on nitric oxide synthase (iNOS), COX-1 and COX-2 protein levels where it showed selectivity for COX-2 compared to COX-1 and iNOS.

NNC 26-9100 increases Aβ1-42 phagocytosis, inhibits nitric oxide production and decreases calcium in BV2 microglia cells.

Microglia are the resident immune cell of the brain involved in the development and progression of Alzheimer's disease (AD). Modulation of microglia activity represents a potential mechanism for treating AD. Herein, the compound NNC 26-9100 (NNC) was evaluated in toxicity, nitric oxide release, Aβ1-42 uptake and cytosolic calcium assays during lipopolysaccharide (LPS)-activated conditions using mouse BV2 microglia cells. After 24 hours, LPS increased cell toxicity in the alamar blue and lactate dehydrogenase assays, increased nitrite release, and increase cytoplasmic calcium. Addition of NNC decreased the LPS-induce lactate dehydrogenase release, had no effect in the alamar blue assay, decreased nitrite release and decreased cytosolic calcium. In the absence of LPS, NNC increased uptake of FITC-tagged Aβ1-42. These data demonstrate that NNC treatment decreases nitrosative stress and microglia cell damage during LPS-induced activation and enhances phagocytosis of Aβ1-42 during non-inflammatory conditions. Thus, NNC 26-9100 may have beneficial effects in AD and in inflammatory diseases of the brain through enhancement of microglial Aβ clearance, and cell protective effects through prevention of elevated cytosolic calcium and inhibition of nitric oxide release.

Leeches alleviates diabetic macroangiopathy by increasing nitric oxide production in rat aorta and restoring endothelium-dependent vasodilatation

IntroductionDiabetes mellitus (DM) is a chronic metabolic disorder characterized by elevated blood glucose level over a prolonged period, leading to severe damage in tissues including the heart, blood vessels, eyes and the kidneys. Danzhi Jiangtang Capsule (DJC) is an effective drug for diabetes, but the mechanism responsible for its efficacy remains unknown. This study aimed to explore the effective ingredient of DJC that ameliorated diabetes and the possible mechanisms.Material and methodsWe orally treated streptozotocin (STZ)-induced diabetic rats with 540 mg/kg DIC or the same dose of its four active components, namely Leeches, Pseudostellaria Polysaccharides (PP), Paeonia Suffruticosa Andr (PSA) and Rehmannia Glutinosa Libosch (RGL), respectively for 8 weeks.ResultsAlthough all of these components could reduce blood glucose levels in diabetic rats, the extent of alleviation of DJC was more pronounced than all of its four ingredients. Unlike the other three components, Leeches is the only effective ingredient of DJC that decreased tetrahydrobiopterin (BH4) oxidation to activate endothelial nitric oxide synthase (eNOS), and increased nitric oxide (NO) production, leading to improved endothelium-dependent relaxation both in diabetic rats and in immortalized human mesangial cells under the stimulation of high glucose.ConclusionsLeeches could alleviate diabetic macroangiopathy by inhibiting NO release in endothelial cells under high-glucose condition.

Potent immunosuppressive and anti-inflammatory bisindole alkaloids from Melodinus fusiformis

Phytochemical investigation of Melodinus fusiformis led to a new aspidosperma-aspidosperma bisindole alkaloid (BIA), bis-19β-hydroxyvenalstonidine (1), together with three known BIAs (2–4). The structures were established by extensive analysis of their HRESIMS, NMR data, and comparing with the reported data. BIA 1 is an almost symmetrical structure, linked by C3–C14’ bond, while BIAs 2–4 are reported for the first time from the plant. The cytotoxic, immunosuppressive and anti-inflammatory activities of BIAs 1–4 were evaluated in vitro. BIAs 1, 3 and 4 showed good toxicity against MOLT-4 cell lines with IC50 values in the range of 1.5–17.5 –M. BIA 2 exhibited the strongest inhibitory effect against MCF-7 cell lines with an IC50 value of 7.1 μM. BIA 1 significantly inhibited Con A-stimulated mice splenocytes proliferation equal to that of the positive control (DXM) in a concentration-dependent manner. BIAs 1 and 2 were able to decrease the NO production in LPS-induced RAW 264.7 cells at 30 μM concentration. BIA 2 showed similar inhibition of nitric oxide release, compared to that of DXM. Furthermore, BIA 2 remarkably inhibited the levels of IL-6 and TNF-α compared to the LPS induced group. Interestingly, BIA 2 displayed an inhibitory effect on TNF-α production similar to that of dexamethasone at a concentration of 20 μM.

Identification of Chemopreventive Components from Halophytes Belonging to Aizoaceae and Cactaceae Through LC/MS-Bioassay Guided Approach.

Six halophytes, namely, Aptenia cordifolia var. variegata, Glottiphyllum linguiforme, Carpobrotus edulis, Ferocactus glaucescens, F. pottsii and F. herrerae were investigated for chemopreventive effect. Prioritization of most promising plant for further investigation was carried out through an integrated liquid chromatography-high resolution electrospray ionization mass spectrometry profiling-bioassay guided approach. NAD(P)H: quinone oxidoreductase-1 (NQO-1) induction in cultured murine hepatoma cells (Hepa-1c1c7) and inhibition of nitric oxide (NO) production in lipopolysaccharide-activated macrophages (RAW 264.7) were carried out to investigate chemopreventive effect. Bioassay data revealed that F. herrerae, A. cordifolia, C. edulis and F. glaucescens were the most active with 2-, 1.7-, 1.6- and 1.5-folds induction of NQO-1 activity. Only F. glaucescens exhibited >50% inhibition of NO release. LCMS profiling of the F. glaucescens revealed its high content of flavonoids, a known micheal acceptor with possible NQO-1 induction, as proved by quantitative high-performance liquid chromatography analysis. Thus, the extract of F. glaucescens was subjected to chromatographic fractionation leading to the isolation of four compounds including (i) 2S-naringenin, (ii) trans-dihydrokaempferol (aromadendrin), (iii) 2S-naringenin-7-O-β-d-glucopyranoside and (iv) kaempferol-7-O-β-d-glucopyranoside (populnin). The current study through an LCMS dereplication along with bio guided approach reported the activity of populnin as NO inhibitor and NQO-1 inducer with promising chemopreventive potential.

Anti-inflammatory canthin-6-one alkaloids from the roots of Thailand Eurycoma longifolia Jack.

Two new canthin-6-one alkaloids, 4,9-dimethoxy-5-hydroxycanthin-6-one (1) and 9-methoxy-(R/S)-5-(1-hydroxyethyl)-canthin-6-one (2), together with fifteen known ones were isolated from the roots of Thailand Eurycoma longifolia Jack. Among the known canthin-6-one alkaloids, compounds 9 and 16 were isolated from the Eurycoma genus for the first time. Meanwhile, the nitric oxide (NO) inhibitory activities of all isolates were examined in lipopolysaccharide (LPS)-stimulated RAW264.7 cells at 50µM. Moreover, a dose-dependent experiment was conducted for active compounds 1, 2, 4, 6, 7, 10, 12-17 at the concentration of 10, 25, and 50µM, respectively. Consequently, compounds 1, 4, 6, 7, 12, 14, 15, as well as 17 were found to inhibit NO release from RAW264.7 cells in a dose-dependent manner. Two new canthin-6-one alkaloids, 4,9-dimethoxy-5-hydroxycanthin-6-one (1) and 9-methoxy-(R/S)-5-(1-hydroxyethyl)-canthin-6-one (2), together with fifteen known ones were isolated from the roots of Thailand Eurycoma longifolia Jack. Among them, 1, 4, 6, 7, 12, 14, 15, as well as 17 were found to inhibit NO release from RAW264.7 cells in a dose-dependent manner at the concentration of 10, 25, and 50 µM.

Thymus zygis subsp. zygis an Endemic Portuguese Plant: Phytochemical Profiling, Antioxidant, Anti-Proliferative and Anti-Inflammatory Activities.

Thymus zygis subsp. zygis is an endemic Portuguese plant belonging to the Thymus zygis species. Although T. zygis is commonly used as a condiment and as a medicinal herb, a detailed description of the polyphenol composition of hydroethanolic (HE) and aqueous decoction (AD) extracts is not available. In this work, we describe for the first time a detailed phenolic composition of Thymus zygis subsp. zygis HE and AD extracts, together with their antioxidant, anti-proliferative and anti-inflammatory activities. Unlike other Thymus species, T. zygis subsp. zygis extracts contain higher amounts of luteolin-(?)-O-hexoside. However, the major phenolic compound is rosmarinic acid, and high amounts of salvianolic acids K and I were also detected. T. zygis subsp. zygis extracts exhibited significant scavenging activity of ABTS+, hydroxyl (•OH), and nitric oxide (NO) radicals. Regarding the anti-proliferative/cytotoxic effect, tested against Caco-2 and HepG2 cells, the AD extract only slightly reduced cell viability at higher concentrations (IC50 > 600 µg/mL, 48 h exposure), denoting very low toxicity, while the HE extract showed a high anti-proliferative effect, especially at 48 h exposure (IC50 of 85.01 ± 15.10 μg/mL and 82.19 ± 2.46 μg/mL, for Caco-2 and HepG2, respectively). At non-cytotoxic concentrations, both extracts reduced the nitric oxide (NO) release by lipopolysaccharide (LPS)-stimulated RAW 264.7 cells (at 50 μg/mL, HE and AD extracts inhibited NO release in ~89% and 48%, respectively). In conclusion, the results highlight the non-toxic effect of aqueous extracts, both resembling the consumption of antioxidants in foodstuff or in functional food. Furthermore, the HE extract of T. zygis subsp. zygis is a source of promising molecules with antioxidant, anti-inflammatory and anticancer activities, highlighting its potential as a source of bioactive ingredients for nutraceutical and pharmaceutical industries.