LPS-activated RAW264 Research Articles

Xylomolins O-X: New limonoids from the Thai mangrove Xylocarpus moluccensis

Ten new limonoids, named xylomolins O-X, were isolated from seeds of the mangrove Xylocarpus moluccensis, collected in the mangrove swamp of Trang Province, Thailand. Their structures were elucidated on the basis of comprehensive spectroscopic data analysis. The absolute configurations of five compounds (1, 3, 8–10) were unequivocally determined by single-crystal X-ray diffraction analyses, conducted with Cu Kα radiation. Xylomolins OU (1–7) are structurally intriguing mexicanolides, and xylomolin V (8) is a derivative of azadirone. Xylomolin W (9) is the first phragmalin 1,8,9-orthoester with report on X-ray crystallography from the genus Xylocarpus. In addition, xylomolin X (10) is the fifth member of the khayalactone class of limonoids with a hexahydro-2H-2,5-propanocyclopenta[b]furan motif. Compounds 1–10 inhibited NO production in LPS-activated RAW 264.7 macrophages in the range of 10.45–95.47% at the concentration of 100.0 μM. Xylomolin X (10) and xylomolin V (8), exhibited the most potent activity with IC50 values of 9.90 ± 1.84 μM and 14.66 ± 2.33 μM, respectively.

Amenyunnaosides A-C, Three New Neolignans Isolated from Amentotaxus yunnanensis and Their Anti-inflammatory Activities.

Phytochemical study on the leaves of Amentotaxus yunnanensis led to the isolation of seventeen phenolic compounds including sixteen neolignans and lignans, and one flavone glycoside. Three among the isolates were previously unreported neolignans and named as amenyunnaosides A-C, respectively. Their structures were elucidated by extensive analyses of HR-ESI-MS, 1D and 2D NMR, and ECD spectra. The isolated neolignans potentially inhibited NO production in LPS-activated RAW264.7 cells with their IC50 values ranging from 11.05 to 44.07 μM, compared to that of the positive control compound, dexamethasone, IC50 value of 16.93 μM. Additionally, amenyunnaoside A dose-dependently reduced production of IL-6 and COX-2 but did not effect to that of TNF-α at concentrations of 0.8, 4, and 20 μM.

Biopharmaceutical Assessment of Mesh Aerosolised Plasminogen, a Step towards ARDS Treatment.

Acute respiratory distress syndrome (ARDS) is a severe complication of lung injuries, commonly associated with bacterial, fungal and viral infections, including SARS-CoV-2 viral infections. ARDS is strongly correlated with patient mortality and its clinical management is very complex, with no effective treatment presently available. ARDS involves severe respiratory failure, fibrin deposition in both airways and lung parenchyma, with the development of an obstructing hyaline membrane drastically limiting gas exchange. Moreover, hypercoagulation is related to deep lung inflammation, and a pharmacological action toward both aspects is expected to be beneficial. Plasminogen (PLG) is a main component of the fibrinolytic system playing key roles in various inflammation regulatory processes. The inhalation of PLG has been proposed in the form of the off-label administration of an eyedrop solution, namely, a plasminogen-based orphan medicinal product (PLG-OMP), by means of jet nebulisation. Being a protein, PLG is susceptible to partial inactivation under jet nebulisation. The aim of the present work is to demonstrate the efficacy of the mesh nebulisation of PLG-OMP in an in vitro simulation of clinical off-label administration, considering both the enzymatic and immunomodulating activities of PLG. Biopharmaceutical aspects are also investigated to corroborate the feasibility of PLG-OMP administration by inhalation. The nebulisation of the solution was performed using an Aerogen® SoloTM vibrating-mesh nebuliser. Aerosolised PLG showed an optimal in vitro deposition profile, with 90% of the active ingredient impacting the lower portions of a glass impinger. The nebulised PLG remained in its monomeric form, with no alteration of glycoform composition and 94% of enzymatic activity maintenance. Activity loss was observed only when PLG-OMP nebulisation was performed under simulated clinical oxygen administration. In vitro investigations evidenced good penetration of aerosolised PLG through artificial airway mucus, as well as poor permeation across an Air-Liquid Interface model of pulmonary epithelium. The results suggest a good safety profile of inhalable PLG, excluding high systemic absorption but with good mucus diffusion. Most importantly, the aerosolised PLG was capable of reversing the effects of an LPS-activated macrophage RAW 264.7 cell line, demonstrating the immunomodulating activity of PLG in an already induced inflammatory state. All physical, biochemical and biopharmaceutical assessments of mesh aerosolised PLG-OMP provided evidence for its potential off-label administration as a treatment for ARDS patients.

Lignan glycosides from Fritillaria verticillata Willd. and their anti-inflammatory activities

Three undescribed lignan glycosides, echiunines E-G (1–3), as well as eight known compounds (4–11) were isolated from Fritillaria verticillata Willd. Among them, compounds 1–3 were a series of lignan glycosides reported for the first time from genus Fritillaria. Their structures were elucidated by analyses of extensive spectroscopic data and comparison of the NMR data with those reported previously, the absolute configuration of compounds were further confirmed by calculated ECD method. The NO release inhibitory effects of compounds were evaluated in LPS-activated RAW264.7 macrophages. Compounds 7–8 showed inhibitory acitivities in a dose-dependent manner.

Diversity-oriented synthesis of diterpenoid alkaloids yields a potent anti-inflammatory agent

BackgroundThe diterpenoid alkaloids belong to a highly esteemed group of natural compounds, which display significant biological activities. It is a productive strategy to expand the chemical space of these intriguing natural compounds for drug discovery. MethodsWe prepared a series of new derivatives bearing diverse skeletons and functionalities from the diterpenoid alkaloids deltaline and talatisamine based on a diversity-oriented synthesis strategy. The anti-inflammatory activity of these derivatives was initially screened and evaluated by the release of nitric oxide (NO), tumor necrosis factor (TNF-α), and interleukin-6 (IL-6) in lipopolysaccharide (LPS)-activated RAW264.7 cells. Futhermore, the anti-inflammatory activity of the representative derivative 31a was validated in various inflammatory animal models, including phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mice ear edema, LPS-stimulated acute kidney injury, and collagen-induced arthritis (CIA). ResultsIt was found that several derivatives were able to suppress the secretion of NO, TNF-α, and IL-6 in LPS-activated RAW264.7 cells. Compound 31a, one of the representative derivatives named as deltanaline, demonstrated the strongest anti-inflammatory effects in LPS-activated macrophages and three different animal models of inflammatory diseases by inhibiting nuclear factor kappa-B (NF-κB)/mitogen-activated protein kinase (MAPK) signaling and inducing autophagy. ConclusionDeltanaline is a new structural compound derived from natural diterpenoid alkaloids, which may serve as a new lead compound for the treatment of inflammatory diseases.

Dimethylbutyrylated Dibenzocyclooctadiene Lignans from the Fruits of Schisandra cauliflora Inhibit NO Production in LPS-Activated RAW264.7 Cells.

From the fruits of Schisandra cauliflora, five new dimethylbutyrylated dibenzocyclooctadiene lignans, named schisandracaurins A-E, were isolated using separation and chromatographic techniques. Their structures were determined by extensive analyses of HR-ESI-MS, NMR, and ECD spectra. The schisandracaurins A-E potentially inhibited NO production in LPS-activated RAW264.7 cells with their IC50 values from 21.4 to 30.3 μM.

Synthesis of new multitarget-directed ligands containing thienopyrimidine nucleus for inhibition of 15-lipoxygenase, cyclooxygenases, and pro-inflammatory cytokines

A new series of thieno[2,3-d]pyrimidine derivatives 4, 5, 6a-o, and 11 was designed and synthesized starting from cyclohexanone under Gewald condition with the aim to develop multitarget-directed ligands (MTDLs) having anti-inflammatory properties against both 15-LOX and COX-2 enzymes. Moreover, the potential of the compounds against the proinflammatory mediators NO, ROS, TNF-α, and IL-6 were tested in LPS-activated RAW 264.7 macrophages. Compound 6o showed the greatest 15-LOX inhibitory effect (IC50 = 1.17 μM) which was superior to that of the reference nordihydroguaiaretic acid (NDGA, IC50 = 1.28 μM); meanwhile, compounds 6h, 6g, 11, and 4 exhibited potent activities (IC50 = 1.29–1.77 μM). The ester 4 (SI = 137.37) and the phenyl-substituted acetohydrazide 11 (SI = 132.26) showed the highest COX-2 selectivity, which was about 28 times more selective than the reference drug diclofenac (SI = 4.73), however, it was lower than that of celecoxib (SI = 219.25). Interestingly, compound 6o, which showed the highest 15-LOX inhibitory activity and 5 times higher COX-2 selectivity than diclofenac, showed a greater poteny in reducing NO (IC50 = 7.77 μM) than both celecoxib (IC50 = 22.89 μM) and diclofenac (IC50 = 25.34), but comparable activity in inhibiting TNF-α (IC50 = 11.27) to diclofenac (IC50 = 10.45 μM). Similarly, compounds 11 and 6h were more potent in reducing TNF-α and IL6 levels than diclofenac, meanwhile, compound 4 reduced ROS, NO, IL6, and TNF-α levels with comparable potency to the reference drugs celecoxib and diclofenac. Furthermore, docking studies for our compounds within 15-LOX and COX-2 active sites revealed good agreement with the biological evaluations. The proposed compounds could be promising multi-targeted anti-inflammatory candidates to treat resistant inflammatory conditions.

Anti-inflammatory and antioxidative effects of gallic acid on experimental dry eye: in vitro and in vivo studies

BackgroundTo investigate the anti-inflammatory and antioxidative effects of gallic acid (GA) on human corneal epithelial cells (HCECs) and RAW264.7 macrophages as well as its therapeutic effects in an experimental dry eye (EDE) mouse model.MethodsA cell counting kit-8 (CCK-8) assay was used to test the cytotoxicity of GA. The effect of GA on cell migration was evaluated using a scratch wound healing assay. The anti-inflammatory and antioxidative effects of GA in vitro were tested using a hypertonic model (HCECs) and an inflammatory model (RAW264.7 cells). The in vivo biocompatibility of GA was detected by irritation tests in rabbits, whereas the preventive and therapeutic effect of GA in vivo was evaluated using a mouse model of EDE.ResultsIn the range of 0–100 μM, GA showed no cytotoxicity in RAW264.7 cells or HCECs and did not delay the HCECs monolayer wound healing within 24 h. Ocular tolerance to GA in the in vivo irritation test was good after seven days. In terms of antioxidative activity, GA significantly reduced the intracellular reactive oxygen species (ROS) in lipopolysaccharide (LPS) activated RAW264.7 macrophages and HCECs exposed to hyperosmotic stress. Furthermore, after pre-treatment with GA, the expression levels of nuclear factor E2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and NADPH quinone oxidoreductase-1 (NQO-1) were significantly upregulated in RAW264.7 macrophages. GA also exhibits excellent anti-inflammatory properties. This is mainly demonstrated by the ability of GA to effectively downregulate the nuclear transcription factor-κB (NF-κB) pathway in LPS-activated RAW264.7 macrophages and to reduce inflammatory factors, such as nitric oxide (NO), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α). In vivo efficacy testing results in a mouse model of EDE showed that GA can effectively prevent and inhibit the apoptosis of corneal epithelial cells (CECs), reduce inflammatory factors in the cornea and conjunctiva as well as protect goblet cells.ConclusionIn vitro and in vivo results indicate that GA possesses potent anti-inflammatory and antioxidative properties with no apparent cytotoxicity within the range of 0–100 μM. It is a promising eye drop formulation for the effective prevention and treatment of dry eye disease (DED).

DJ-X-013 ameliorates inflammation by reducing TNF-α/NF-κB expression and experimental colitis by inducing myeloid-derived suppressor cells

Abstract Inflammation is a body’s defense response for a short duration however if persists for long periods, it causes tissue damage and pathological consequences. Moreover, it paves the way for several diseases including inflammatory bowel disease (IBD). Therefore, there is an urgent need for a natural and safe anti-inflammatory therapeutic compound. To address this issue, we aimed to determine the anti-inflammatory effects of DJ-X-013, using lipopolysaccharide (LPS) activated RAW 264.7 macrophage in vitro and dextran sodium sulfate (DSS) induced experimental model of colitis in vivo. DJ-X-013 drastically reduced tumor necrosis factor-alpha and inducible nitric oxide synthase expression in LPS-activated RAW 264.7 macrophages. Further, DJ-X-013 also impeded the expression of NF-κB and other inflammatory markers and pathways like IL-6, IL-1β, STAT3, ERK, and JNK as compared to LPS-activated cells. Additionally, DJ-X-013 also modulates the migratory activity of RAW 264.7 macrophages during wound healing. Next, using DSS induced colitis model, we observed that the reduction of body weight and colon length were slightly improved in the DJ-X-013 group compared to the control. Intriguingly, myeloid-derived suppressor cells, neutrophils, and CD8 +T cells increased, while Th17 cells frequency decreased in the spleen, mesenteric lymph nodes, and colon lamina propria in DJ-X-013 treated group as compared to DSS. However, natural killer and natural killer T cells were marginally enhanced in the DJ-X-013 group as compared to the DSS alone. Hence, the DJ-X-013 exhibits an immunomodulatory effect and might be a promising drug for IBD and other inflammatory diseases, however, further investigation is required for prudent conclusions. This research is supported by NIH grant R01 AI140405

An integrated approach to the structural characterization, long-term toxicological and anti-inflammatory evaluation of Pterospermum rubiginosum bark extract.

Pterospermum rubiginosum is an evergreen plant in Western Ghats, India, used by traditional tribal healers due to its excellent biological potential for treating inflammation and pain relief procedures. The bark extract is also consumed to relieve the inflammatory changes at the bone fractured site. The traditional medicinal plant in India have to be characterized for its diverse phytochemical moieties, its interactive multiple target sites, and to reveal the hidden molecular mechanism behind the biological potency. The study focussed on plant material characterization, computational analysis (prediction study), toxicological screening (In vivo), and anti-inflammatory evaluation of P. rubiginosum methanolic bark extracts (PRME) in LPS-induced RAW 264.7cells. The pure compound isolation of PRME and their biological interactions were used to predict the bioactive components, molecular targets, and molecular pathways of PRME in inhibiting inflammatory mediators. The anti-inflammatory effects of PRME extract were evaluated in the lipopolysaccharide (LPS)-induced RAW264.7 macrophage cell model. The toxicity evaluation of PRME was performed in healthy 30 Sprague-Dawley experimental rats, were randomly divided into five groups for toxicological evaluation for 90 days. The tissue levels of oxidative stress and organ toxicity markers were measured using the ELISA method. Nuclear magnetic resonance spectroscopy (NMR) was performed to characterize the bioactive molecules. Structural characterization revealed the presence of vanillic acid, 4-O-methyl gallic acid, E-resveratrol, gallocatechin, 4'-O-methyl gallocatechin, and catechin. Molecular docking of NF-kB exhibited significant interactions with vanillic acid and 4-O-methyl gallic acid with binding energy -351.159 Kcal/Mol and -326.5505 Kcal/Mol, respectively. The PRME-treated animals showed an increase in total GPx and antioxidant levels (SOD and catalase). Histopathological examination revealed no variation in the liver, renal and splenic tissue's cellular pattern. PRME inhibited the pro-inflammatory parameters (IL-1β, IL-6, and TNF-α) in LPS-induced RAW 264.7cells. The protein level of TNF-α and NF-kB protein expression study brought out a notable reduction and exhibited a good correlation with the gene expression study. The current study establishes the therapeutic potential of PRME as an effective inhibitory agent against LPS-activated RAW 264.7cells induced inflammatory mediators. Long-term toxicity evaluation on SD rats confirmed the non-toxic nature of PRME up to 250mg/body weight for 3 months.

Network Pharmacology Study of Bioactive Components and Molecular Mechanisms of the Glycoside Fraction from Picrorhiza scrophulariiflora Against Experimental Colitis.

To explore the potential mechanism of glycosidic fraction of Picrorhiza scrophulariiflora Pennell (GPS) extract for the treatment of colitis using UPLC-QTOF-MS analysis, network pharmacology and experimental research. The active components of GPS extract were identified by UPLC-QTOF-MS analysis and extracted their targets from the databases, which was used for network pharmacology analysis. Kyoto Encyclopedia of genes and genomes (KEGG) pathway analysis was performed to discover potential therapeutic mechanisms, and the network pharmacology results were then validated by in vivo and in vitro experiments. The results showed that GPS extract significantly alleviated the clinical signs of colitis, including body weight, disease activity index, colon shortening, and colon tissue damage, and inhibited the transcription and production of colonic IL-1β and IL-6 in DSS-induced colitis mice. In vitro, GPS extract also significantly suppressed nitric oxide (NO) production, iNOS expression, IL-1β and IL-6 transcription of LPS-activated RAW 264.7 cells. Network pharmacology integrated with experimental validation identified that GPS extract significantly suppressed Akt, p38, ERK, and JNK phosphorylation in vivo and in vitro, and luteolin, apocynin, caffeic acid, caffeic acid methyl ester, luteoloside, picroside II, aucubin, cinnamic acid, vanillic acid, and sweroside were the main components responsible for the anti-inflammatory effect of GPS. These findings demonstrate that the potential anti-inflammatory effect of GPS extract against colitis is achieved through suppressing PI3K/Akt and MAPK pathways, and that the abovementioned active components mainly exerted its anti-inflammatory effect. The therapeutic effect of GPS extract on colitis is related to PI3K/Akt and MAPK pathways, which is a promising remedy for colitis therapy.

Anti-inflammatory lanostane triterpenoids with rearranged spirobi[indene] scaffold and their biogenetically related analogues from Euphorbia maculata

Phytochemical investigations on the ethanol extract of the whole plant of Euphorbia maculata Linn. Resulted in the identification of 16 lanostane-related triterpenoids, including 11 undescribed ones, namely spiromaculatols A–C (1–3) and euphomaculatoids A–H (4–11). The structural determinations of the previously undescribed ones (1–11) were elucidated based on the interpretation of comprehensive spectroscopic data, quantum chemical calculation, as well as X-ray crystallographic experiments. Spiromaculatols A–C (1–3) possess a rare spirobi [indane] skeleton, which was biosynthetically derived from the 7 (8 → 9)-abeo bond migration of lanostane precursors. The biological activity of compounds 1–3, 5, 7, and 12–13 displayed inhibitory effect on the release of NO in an LPS-activated RAW264.7 cells model. Molecular mechanism study indicated that the most potent spiromaculatol C (3) can reduce the nuclear translocation of NF-κB p65 and decrease the transcriptional expressions of its downstream pro-inflammatory mediators.

Natriuretic-like Peptide Lebetin 2 Mediates M2 Macrophage Polarization in LPS-Activated RAW264.7 Cells in an IL-10-Dependent Manner.

Snake natriuretic peptide (NP) Lebetin 2 (L2) has been shown to improve cardiac function and reduce fibrosis as well as inflammation by promoting M2-type macrophages in a reperfused myocardial infarction (MI) model. However, the inflammatory mechanism of L2 remains unclear. Therefore, we investigated the effect of L2 on macrophage polarization in lipopolysaccharide (LPS)-activated RAW264.7 cells in vitro and explored the associated underlying mechanisms. TNF-α, IL-6 and IL-10 levels were assessed using an ELISA assay, and M2 macrophage polarization was determined by flow cytometry. L2 was used at non-cytotoxic concentrations determined by a preliminary MTT cell viability assay, and compared to B-type natriuretic peptide (BNP). In LPS-activated cells, both peptides reduced TNF-α and IL-6 release compared to controls. However, only L2 increased IL-10 release in a sustained manner and promoted downstream M2 macrophage polarization. Pretreatment of LPS-activated RAW264.7 cells with the selective NP receptor (NPR) antagonist isatin abolished both IL-10 and M2-like macrophage potentiation provided by L2. In addition, cell pretreatment with the IL-10 inhibitor suppressed L2-induced M2 macrophage polarization. We conclude that L2 exerts an anti-inflammatory response to LPS by regulating the release of inflammatory cytokines via stimulating of NP receptors and promoting M2 macrophage polarization through activation of IL-10 signaling.

Chromatographic Determination of the Absolute Configuration in Sanjoinine A That Increases Nitric Oxide Production.

A chiral derivatization strategy with phenylglycine methyl ester (PGME) was employed to develop a straightforward method to determine the absolute configurations of N,N-dimethyl amino acids. The PGME derivatives were analyzed using liquid chromatography-mass spectrometry to identify the absolute configurations of various N,N-dimethyl amino acids based on their elution time and order. The established method was applied to assign the absolute configuration of the N,N-dimethyl phenylalanine in sanjoinine A (4), a cyclopeptide alkaloid isolated from Zizyphi Spinosi Semen widely used as herbal medicine for insomnia. Sanjoinine A displayed production of nitric oxide (NO) in LPS-activated RAW 264.7 cells.

Investigation of Cajuput and Lemongrass Essential Oils Supplemented in Alcohol-Free Mouthwash: Anti-Inflammation on Human Gingival Fibroblast Cells In Vitro

Alcohol-free mouthwash containing essential oils is a helpful adjunct to self-performed oral hygiene of patients with periodontal disease due to its antimicrobial and anti-inflammatory properties. This study aimed to investigate the anti-inflammatory activity of cajuput and lemongrass essential oils supplemented in alcohol-free mouthwash on human gingival fibroblast cells and RAW 264.7 macrophages, which were incubated in a complete medium. In the current in vitro study, 3 types of mouthwash, including 12 % (v/v) alcohol-free mouthwash, 0.8 % (v/v) cajuput essential oil and 0.4 % (v/v) lemongrass essential oil supplemented in alcohol-free mouthwash, and 0.12 % chlorhexidine mouthwash, were evaluated. All 3 types were determined for cytotoxicity by MTT assay. The sub-IC50 concentration of each mouthwash was calculated to investigate anti-inflammatory activities via the inhibition of lipopolysaccharide-activated nitric oxide production, cyclooxygenases-2, interleukin-1β and interleukin-6 gene expression. The wound healing rate was determined by scratch assay. The results demonstrated that the sub-IC50 concentrations of essential oils in 0.8 % (v/v) cajuput essential oil and 0.4 % (v/v) lemongrass essential oil supplemented in alcohol-free mouthwash significantly inhibited nitric oxide production in lipopolysaccharide-activated RAW 264.7 macrophages and reduced the gene expression of cyclooxygenases-2, interleukin-1β and interleukin-6 in lipopolysaccharide-activated human gingival fibroblast cells, while stimulating the migration of human gingival fibroblast cells, similar to the sub-IC50 concentration of 0.12 % chlorhexidine mouthwash. These findings suggested that essential oils contained in this mouthwash formula exhibited an anti-inflammatory effect and promoted oral wound healing, which might be used as an alternative agent for patients with periodontal disease. HIGHLIGHTS Cajuput and lemongrass essential oils supplemented in alcohol-free mouthwash expressed anti-inflammatory activity by inhibiting nitric oxide production in LPS-activated RAW 264.7 macrophages and reducing COX-2, IL-6 and IL-1β expression in LPS-activated HGFs, as well as by promoting oral wound healing, similar to chlorhexidine mouthwash, which is a common, well-known agent in dental treatment Our findings suggested that this novel mouthwash may be an alternative agent in preventing oral inflammatory diseases, especially periodontal disease GRAPHICAL ABSTRACT

Withanolides from Physalin angulata and their inhibitory effects on nitric oxide production.

Six new withanolides, angulasteroidins A-F (1-6), along with twelve known analogues (7-18) were isolated from the whole plants ofPhysalisangulata. Their structures were elucidated by analysis of 1D and 2D NMR, ECD spectra and HRESIMS data.Compounds 2-4, 7-9, and 15 exhibited significantly inhibitory activity on the production of nitric oxide in the LPS-activated RAW 264.7 mouse macrophage cell line with IC50values ranging from 0.23 to 9.06 μM.Six new withanolides, angulasteroidins A-F (1-6), along with twelve known analogues (7-18) were isolated from the whole plants of Physalis angulata . Their structures were elucidated by analysis of 1D and 2D NMR, ECD and IR spectra, HRESIMS data, and ECD calculation.Compounds 1 and 6 were rare 1-10 seco withanolides. Compounds 2-4, 7-9, and 15 exhibited significant inhibitory activity on the production of nitric oxide in the LPS-activated RAW 264.7 mouse macrophage cell line with IC50values ranging from 0.23 to 9.06 μM.

In Vitro Anti-inflammatory Activity of Three Inula Species Essential Oils in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages

In the face of the undesirable effects induced by anti-inflammatory drugs, there has been a return, nowadays, to the search for active ingredients based on plants. Herein, for the first time we study the anti-inflammatory activity of essential oils of three species of the genus Inula: Inula viscosa, Inula graveolens and Inula crithmoides in lipopolysaccharide (LPS)-activated macrophages. Essential oils have shown excellent preventive anti-inflammatory potential by causing inhibition of nitric oxide (NO) production in LPSactivated RAW264.7 macrophages with IC50s ranging between 15 and 35 µg mL− 1. On the other hand, the major acidic compounds, more precisely α- and β-costic acids, have been isolated from Inula viscosa and Inula graveolens essential oils and evaluated for their anti-inflammatory effect. These compounds appear to have a moderate preventive inhibitory effect on NO production relative to the significant effect generated by the neutral minority components present in the oils such as borneol, bornyl acetate, (E)-nerolidol, caryophyllene oxide, T-cadinol and eugenol. Therefore, we can deduce that the studied essential oils could be used as anti-inflammatory agents for the treatment of various inflammatory pathologies.

Chemical Constituents of Thesium chinense Turcz and Their In Vitro Antioxidant, Anti-Inflammatory and Cytotoxic Activities.

Three novel compounds (1-3) along with twenty-six known compounds, two known steroids (4-5) and twenty-four known phenylpropanoids (6-29) were isolated from the whole plant of Thesium chinense Turcz. The structures of the three new compounds were elucidated on the basis of ESI-MS, HR-ESIMS, 1D and 2D NMR, IR, UV spectroscopic data. The absolute stereochemistry of compound 1 was determined by the Gauge-Including Atomic Orbitals (GIAO) method. The in vitro antioxidant, anti-inflammatory and cytotoxic activities of the isolated compounds were evaluated by DPPH radical-scavenging assay, LPS-activated RAW 264.7 cells model and CCK-8 kit, respectively. Compound 11 showed high antioxidant activity with an SC50 value of 16.2 ± 1.6 μM. Compound 21 showed considerable anti-inflammatory activity with an IC50 value of 28.6 ± 3.0 μM. Compounds 4 and 5 displayed potent cytotoxic activity against human NCI-H292, SiHa, A549, and MKN45 cell lines, with the compound 4 having IC50 values of 17.4 ± 2.4, 22.2 ± 1.1, 9.7 ± 0.9, 9.5 ±0.7 μM, and the compound 5 having all IC50 values less than 0.1 μM in vitro.

Polyene Carboxylic Acids from a Streptomyces sp. Isolated from Tibet Soil.

Six new polyene carboxylic acids named serpentemycins E-J (1-6), together with three known analogs (7-9), were isolated from the fermentation medium of Streptomyces sp. TB060207, which was isolated from arid soil collected from Tibet, China. The structures of the new compounds were elucidated mainly on the basis of HR-ESI-MS and NMR spectroscopic analyses. The inhibitory activities of compounds 1-9 against NO production in LPS-activated RAW264.7 cells were evaluated. Compound 9 has an inhibition rate of 87.09% to 60.53% at concentrations ranging from 5.0 to 40.0 µM.

Cyclic pentapeptides with anti-inflammatory, cytotoxic or α-glucosidase inhibitory activities from Basidiobolus meristosporus

Basidiobolus meristosporus is an opportunistic pathogen of mammals with unique habitats, but its metabolites have not been extensively studied. Through semi-preparative HPLC, nine undescribed cyclic pentapeptides were isolated from mycelia of B. meristosporus RCEF4516. The structure of the compounds 1–9 were identified with MS/MS and NMR data and designated as basidiosin D-L respectively. The absolute configurations were determined according to the advanced Marfey's method after compound hydrolysis. Bioactivity testing showed that compounds 1, 2, 3, 4 and 8 decreased NO production in LPS-activated RAW264.7 cells in a concentration-dependent manner. The nine compounds showed cytotoxicity against RAW264.7, 293 T and HepG2 cells. All the compounds except compound 7 showed stronger inhibitory effects on α-glucosidase than acarbose.