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Mtb (Mycobacterium tuberculosis) is a pathogenic bacterium that causes tuberculosis infection (TB). Mtb -secreted proteins have recently been investigated as virulence factors, as well as therapeutic and vaccine possibilities. The early-secreted antigen target MTB48 is one of these proteins that has been explored as a cocktail antigen in the serodiagnosis of active tuberculosis. However, there exists no information about the function or control of MTB48's inflammatory activity in macrophages at the site of inflammation. As a result, the goal of this research was to figure out what processes are involved in MTB48's function. MTB48 stimulated inflammation in LPS induced macrophages at both the protein and mRNA levels, which was interesting. MTB48 aided LPS induced IB phosphorylation and NF-κB translocation. MTB48 also led to the phosphorylation of MAPK signaling protein. These findings imply that MTB48 can enhance inflammatory activity via NF-κB and MAPK signaling by upregulating COX-2, iNOS, NO and PGE 2. Many tuberculosis antigens have been tested for the development of rapid serological diagnosis. The results of this study suggest that MTB48 is a very high conservative antigen and is a major factor causing inflammatory reactions, suggesting that it can help control and diagnose tuberculosis. [Display omitted] • The function and control of the Mtb-secreted protein MTB48's inflammatory activity in macrophages at the site of inflammation. • MTB48 enhanced inflammatory activity via NF-κB and MAPK signaling by upregulating COX-2, iNOS, NO, and PGE2. • MTB48 could be a potential target for therapeutic and vaccine possibilities, as well as a diagnostic tool for active tuberculosis. • MTB48 is a highly conservative antigen and a major factor in causing inflammatory reactions. [ABSTRACT FROM AUTHOR]

[Display omitted] The potentiation of pharmacological effects can be achieved through several strategies, such as the association of substances and delivery in nanostructured systems. In practice, potentiation can be measured by the law of mass action and joint evaluation of the combination index (CI) and dose–response curves. In this context, this study aimed to evaluate the anti-inflammatory effect of the association of β-caryophyllene and indomethacin in the free form and delivered in nanoemulsions using the in vitro model of LPS-stimulated murine macrophage. The results indicated potentiation of the anti-inflammatory effect of nanoemulsified substances compared to free substances, as well as synergistic action between the sesquiterpene and the selected NSAID. In comparison, the association of β-caryophyllene and indomethacin in the free form inhibited the production of nitric oxide by 50% at 48.60 µg/mL (CI = 0.21), while the nanoemulsified association of these substances resulted in an IC 50 of 1.45 µg/mL (CI = 0.14). In parallel, cytotoxicity assays on HaCaT and MRC-5 cell lines demonstrated the safety of IC 50 -equivalent concentrations of the anti-inflammatory action, and no irritating effects on the chorioallantoic membrane of embryonated eggs were observed (HET-CAM assay). The results suggest that β-caryophyllene may be an alternative to replace an inert oily core in nanoemulsion systems when anti-inflammatory effects are desirable. [ABSTRACT FROM AUTHOR]

Conventionally T-cell receptors (TCRs) have so far been considered as a T-lymphocyte privilege. However, recent findings also place TCR expression in non-lymphoid cells, namely neutrophils, eosinophils and macrophages. In order to examine the ectopic expression of TCR, this study focused on RAW 264.7 cells, which have been broadly used for their macrophage properties. Immunofluorescence staining detected 70% and 40% of the cells to express TCRαβ and TCRγδ respectively, which was also verified by RT-PCR experiments and confocal microscopy analysis. Interestingly, except from the predicted 292 and 288 bp gene products for the α- and γ-chain, additional products at 220 and 550 bp could be detected, respectively. RAW 264.7 cells also expressed the co-stimulatory CD4 and CD8 markers at a percentage of 61% and 14% respectively, which supported the expression of TCRs. However, only low numbers of cells expressed CD3ε and CD3ζ (9% and 7% respectively). Such observations contradicted the existing knowledge, and indicated that TCRs would be supported by other molecules for reaching the membrane and transducing their signal. Such candidate molecules could be the Fcγ receptors (FcγRs). Indeed, the FcγRII/III receptor was found to be expressed in 75% of the cells, which also expressed at a percentage of 25% major histocompatibility complex (MHC) class II molecules. Engagement of the FcγRII/III receptor by a recombinant IgG2aCH2 fragment, except from stimulating the macrophage-dependent properties of the cells, was shown to reduce expression of TCRαβ and γδ indicating that FcγRII/III was indeed used by TCRs for their transport to the cell membrane. In order to examine the ability of RAW 264.7 cells to simultaneously display antigen presenting- and T-cell properties, functional experiments as to antigen-specific antibody and IL-2 production were performed. In in vitro immunization assays in the presence of naïve B cells, RAW264.7 failed to promote antibody production. However, RAW 264.7 cells could compete with antigen-stimulated macrophages but not T cells when applied to a system of in vivo antigen-sensitized cells followed by an in vitro immunization protocol. Interestingly, simultaneous addition of antigen and the IgG2aCH2 fragment to RAW 264.7 cells could promote IL-2 production from the cells, indicating that FcγRII/III activation could also support TCR stimulation. Extrapolating these findings to cells of the myeloid origin, the above results dictate novel regulatory mechanisms towards the alteration of the immune response. [Display omitted] • RAW 264.7 macrophages express TCRαβ/TCRγδ and CD4/CD8 but insufficient CD3 levels. • Recombinant IgG2aCH2 decreases TCRαβ/TCRγδ indicating that FcγRII/III is involved in TCR membrane transport. • RAW 264.7 cells may compete macrophages but not T cells in an antigen specific antibody production. • Stimulation of RAW 264.7 cells with rIgG2aCH2 and antigen induces IL-2 production. • Novel regulatory pathways by TCR+ myeloid cells towards stimulation of immunity. [ABSTRACT FROM AUTHOR]

It has been reported that 4,5-dihydropyrazole and thiazole derivatives have many biological functions, especially in the aspect of anti-inflammation. According to the strategy of pharmacophore combination, we introduced thiazolinone and dihydropyrazole moiety into steroid skeleton to design and synthesize a novel series of D-ring substituted steroidal 4,5-dihydropyrazole thiazolinone derivatives, and assessed their in vitro anti-inflammatory profiles against Lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells. The anti-inflammatory activities assay demonstrated that compound 12e was considered as the most effective anti-inflammatory drug, which suppressed the expression of pro-inflammatory mediators including nitric oxide (NO), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α), it also dose-dependently inhibited the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in LPS-induced RAW 264.7 macrophage cells. Furthermore, the results of the Western blot analysis showed a correlation between the inhibition of the Nuclear factor-kappa B (NF-κB) and Mitogen-activated protein kinases (MAPKs) signaling pathways and the suppressive effects of compound 12e on pro-inflammatory cytokines. Molecular docking studies of compound 12e into the COX-2 protein receptor (PDB ID: 5IKQ) active site was performed to rationalize their COX-2 inhibitory potency. The results were found to be in line with the biological findings as they exerted more favorable interactions compared to that of dexamethasone (DXM), explaining their remarkable COX-2 inhibitory activity. The findings revealed that these candidates could be identified as potent anti-inflammatory agents, compound 12e could be a promising drug for the treatment of inflammatory diseases. [Display omitted] • This study provided a novel, efficient and simple synthesis of steroidal 4,5-dihydropyrazole thiazolinone derivatives. • In vitro activity assay screened compound 12e with excellent anti-inflammatory biological activity and low cytotoxicity. • 12e inhibited LPS-induced expression of the pro-inflammatory cytokines iNOS, COX-2, TNF-α and IL-6 on RAW 264.7 cells. • 12e exerted anti-inflammatory effects through down-regulation of NF-κB/MAPKs pathway. [ABSTRACT FROM AUTHOR]

Inflammation, an important biological protective response to tissue damage or microbial invasion, is considered to be an alarming signal for the progress of varied biological complications. Based on the previous reports in the literature that proved the noticeable efficacy of pyrazole and thiazole scaffold as well as nitrogen heterocyclic based compounds against acute and chronic inflammatory disease, a new set of novel D-ring substituted steroidal 4,5-dihydropyrazole thiazole derivatives were synthesized and evaluated their anti-inflammatory activities in vitro. Preliminary structure-activity relationship (SAR) analysis was conducted by their inhibitory activities against nitric oxide (NO) release in lipopolysaccharide (LPS)-induced RAW 264.7 cells, and the optimal compound 12b [3β-hydroxy-pregn-5-en-17β-yl-5′- (o- chlorophenyl)− 1′-(4′′- phenyl -[1′′, 3′′]- thiazol-2′′- yl) − 4′,5′-dihydro − 1′H-pyrazol − 3′- yl] exhibited more potent anti-inflammatory activity than the positive control treatment methylprednisolone (MPS), with an IC 50 value of 2.59 μM on NO production and low cytotoxicity against RAW 264.7 cells. In further mechanism study, our results showed that compound 12b significantly suppressed the production of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and inhibited the expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) through blocking NF-κB p65 nuclear translocation and phosphorylation of IκBα. Compound 12b also attenuated LPS-induced activation of c-Jun amino-terminal kinase (JNK) and p38 phosphorylation in RAW 264.7 cells. Molecular docking study revealed the strong binding affinity of compound 12b to the active site of the COX-2 proteins, which confirmed that compound 12b acted as an anti-inflammatory mediator. These results indicate that steroidal derivatives bearing 4,5-dihydropyrazole thiazole structure might be considered for further research and scaffold optimization in designing anti-inflammatory drugs and compound 12b might be a promising therapeutic anti-inflammatory drug candidate. [Display omitted] • This study provided a novel series of D-ring substituted steroidal 4,5-dihydropyrazole thiazole derivatives. • The structures of all targeted synthesized compounds had been characterized. • The anti-inflammatory biological activity of targeted synthesized compounds was assayed. • In vitro activity assay screened compound 12b with excellent anti-inflammatory biological activity in LPS-induced RAW 264.7 cells. [ABSTRACT FROM AUTHOR]

[Display omitted] • Non-starch polysaccharide (ECPs-I) was extracted from finger millet. • Deproteinated non-starch polysaccharide (ECPs-II) prepared by proteinase K. • The molecular weight of polysaccharides were found to range from 43 to 120 kDa. • The extracted polysaccharides did not show any toxic effects on RAW 264.7 cells. • ECPs-II has significantly enhance the RAW 264.7 cells activation. The structural elucidation and investigation of the immunostimulatory effects of crude polysaccharides (ECPs), non-starch polysaccharides (ECPs-I), and deproteinated non-starch polysaccharides (ECPs-II) derived from finger millet (Eleusine coracana) were performed. The extracted ECPs, ECPs-I, and ECPs-II primarily comprised different levels of carbohydrate (82.9, 41.9 and 62.0%), protein (5.3, 38.4 and 22.0%) and uronic acid (0.5, 2.4 and 0.7%). Monosaccharide composition analysis showed that glucose (88.2 and 40.6) was the main sugar unit present in ECPs and ECPs-I, followed by galactose, arabinose, xylose, rhamnose, and mannose; however, ECPs-II contained galactose (42.2%) as a major unit. The average molecular weights of ECPs, ECPs-I, and ECPs-II were found to be 120, 78 and 43 kDa, respectively. The polysaccharides as well as non-starch polysaccharides did not show any toxic effects on RAW 264.7 cells. ECPs-II treatment significantly enhanced the activation of RAW 264.7 cells by inducing nitric oxide (NO) production, upregulating inducible nitric oxide synthase (iNOS) and various pro-inflammatory cytokines including interleukin (IL)-1β, IL-6, IL-10, and tumor necrosis factor alpha (TNF-α). Furthermore, the RAW 264.7 cells were activated after ECPs-II treatment via the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), and mitogen-activated protein kinase signaling (MAPK) pathways induced by a cluster of differentiation 40 (CD40) expression. The structure of ECPs-II mainly consists of (1→3,4)-linked galactopyranosyl and (1→3,5) arabinopyranosyl. [ABSTRACT FROM AUTHOR]

The usefulness of sirolimus (SIR) in the treatment of diseases that involve retinal degeneration like age-related macular degeneration (AMD) has been well documented. However, the problem still remains probably owing to the peculiar environment of the eye and/or unfavourable physiochemical profile of SIR. In the present work, we aimed to fabricate sirolimus loaded PLGA nanoparticles (SIR-PLGA-NP) and chitosan decorated PLGA nanoparticles (SIR-CH-PLGA-NP) to be administered via non-invasive subconjunctival route. Both the nanoparticles were characterized in terms of size, zeta potential, DSC, FTIR and XRD analysis. Quality by Design (QbD) approach was employed during the preparation of nanoparticles and the presence of chitosan coating was confirmed through thermogravimetric analysis and contact angle studies. Cationic polymer modification showed sustained in-vitro SIR release and enhanced ex-vivo scleral permeation and penetration. Further, SIR-CH-PLGA-NP revealed enhanced cellular uptake and thus, reduced lipopolysaccharide (LPS)-induced free-radicals generation by RAW 264.7 cells. The prepared nanoparticles were devoid of residual solvent and were found to be safe in HET-CAM analysis, RBCs damage analysis and histopathology studies. Moreover, high anti-angiogenic potential was observed in SIR-CH-PLGA-NP compared with SIR-PLGA-NP in chorioallantoic membrane (CAM) test. Overall, the current work opens up an avenue for further investigation of CH-PLGA-NP as SIR nanocarrier in the treatment of AMD. [Display omitted] • Quality by Design (QbD) approach was employed in preparation of nanoparticles. • Sirolimus loaded PLGA nanoparticles were successfully functionalized with chitosan. • Chitosan functionalization enhanced cellular uptake and scleral penetration. • SIR-CH-PLGA nanoparticles were safe with higher anti-angiogenic potential. [ABSTRACT FROM AUTHOR]

Oldenlandia umbellata L., belonging to the Rubiaceae family, is an annual plant possessing anti-inflammatory and antipyretic, anti-nociceptive, anti-bacterial, anti-helminthic, antioxidant and hepatoprotective activities and used in traditional medicine to treat inflammation and respiratory diseases. The present study aims to evaluate the anti-osteoporotic effect of Methanolic extract of O.umbellata in MG-63 cells and RANKL-stimulated RAW 264.7 cells. The methanolic extract from the aerial parts of O.umbellata was subjected to metabolite profiling. The anti-osteoporotic effect of MOU was assessed in MG-63 cells and RANKL-stimulated RAW 264.7 cells. In MG-63 cells, the proliferative effect of MOU was evaluated using MTT assay, ALP assay, Alizarin red staining, ELISA and western blot. Similarly, the anti-osteoclastogenic effect of MOU was assessed in RANKL-stimulated RAW 264.7 cells via MTT, TRAP staining and western blot. LC-MS metabolite profiling showed the presence of 59 phytoconstituents including scandoside, scandoside methyl ester, deacetylasperuloside, asperulosidic acid, and cedrelopsin in MOU. In MG-63 cells, MOU has increased the proliferation of osteoblast cells and ALP activity, thereby increasing bone mineralization. ELISA results showed increased levels of osteogenic markers such as osteocalcin and osteopontin in the culture media. Western blot analysis showed inhibition of GSK3β protein expression and increased the expression levels of β-catenin, Runx-2, col 1 and osterix, promoting osteoblast differentiation. In RANKL-stimulated RAW 264.7 cells, MOU did not elicit any significant cytotoxicity; instead, it suppressed the osteoclastogenesis reducing the osteoclast number. MOU has reduced TRAP activity in a dose-dependent manner. MOU inhibited the TRAF6, NFATc1, c-Jun, C-fos and cathepsin K expression, thereby inhibiting osteoclast formation. In conclusion, MOU promoted osteoblast differentiation via inhibiting GSK3β and activating Wnt/β catenin signalling and its transcription factors, including β catenin, Runx2 and Osterix. Similarly, MOU inhibited osteoclast formation by inhibiting the expression of TRAF6, NFATc1, c-Jun, C-fos and cathepsin K in RANK-RANKL signalling. Finally, it can be emphasised that O.umbellata is a potential source of therapeutic leads for the treatment of osteoporosis. [Display omitted] [ABSTRACT FROM AUTHOR]

The neonicotinoid imidacloprid was promoted in the market because of widespread resistance to other insecticides, plus its low mammalian impact and higher specific toxicity towards insects. This study aimed to evaluate the immunomodulatory effect of imidacloprid on macrophages. RAW 264.7 cells were incubated to 0–4000 mg/L of imidacloprid for 24 and 96 h. Imidacloprid presented a concentration-dependent cytotoxicity after 24 h and 96 h incubation for MTT reduction (3-(4,5-dimethyl-thiazol-2-yl)− 2,5-diphenyltetrazolium bromide) (EC 50 519.6 and 324.6 mg/L, respectively) and Neutral Red (3-amino-7-dimethylamino-2-methylphenazine hydrochloride) assays (EC 50 1139.0 and 324.2 mg/L, respectively). Moreover, imidacloprid decreased the cells' inflammatory response and promoted a mitochondrial depolarization. The complex II and succinate dehydrogenase (SDH) activities in RAW 264.7 cells incubated with imidacloprid increased more at 24 h. These results suggest that imidacloprid exerts an immunomodulatory effect and mitochondria can act as regulator of innate immune responses in the cytotoxicity mediated by the insecticide in RAW 264.7 cells. [Display omitted] • A commercial formulation of imidacloprid impaired the inflammatory response in RAW 264.7 cells; • The immunotoxic effects were associated with long term exposure to imidacloprid; • The mitochondria is a primary target of imidacloprid cytotoxicity. [ABSTRACT FROM AUTHOR]

• A new family of D-ring pyrazoline derivatives of pregn-5-enes were synthesized. • The structures of all targeted synthesized compounds had been characterized. • The anti-inflammatory biological activity of synthesized compounds was assayed. • Compound 4g was with excellent anti-inflammatory activity and low cytotoxicity. To identify new potential anti-inflammatory agents, a number of novel steroidal derivatives with nitrogen heterocyclic side chains 4a-4l were synthesized and evaluated for their anti-inflammatory effects in activated RAW 264.7 macrophage cells. The synthesis scheme involves two steps, Claisen-Schmidt condensation with the corresponding pregnenolone and aromatic aldehydes as the first step followed by nucleophilic addition of thiosemicarbazide across an α, β-unsaturated carbonyl as a later step. Compound structures were confirmed by 1H NMR, 13C NMR, HRMS, and IR. The compounds were assayed to test their anti-inflammatory effects in activated RAW 264.7 cells. Compound 4g , 3β-hydroxy-pregn-5-en-17β-yl-5′-(m-fluorophenyl)-4′, 5′-dihydro-1′-carbothioic acid amido pyrazole, was identified as the most potent anti-inflammatory agent of the analysed compounds, with an IC 50 value of 0.86 µM on nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW 264.7 cells for 24 h compared to dexamethasone (IC 50 = 0.62 µM) and low cytotoxicity against RAW 264.7 cells. Compound 4g significantly inhibited NO produced by LPS-induced RAW 264.7 cells. Further studies showed that compound 4g markedly inhibited the expression of pro-inflammatory factors, including inducible nitric oxide synthase (iNOS), interleukin-6 (IL-6), tumour necrosis factor-α (TNF-α), and cyclooxygenase-2 (COX-2) in LPS-induced RAW 264.7 cells. These results indicate that derivatives bearing pyrazoline structure might be considered for further research and scaffold optimization in designing anti-inflammatory drugs and compound 4g might be a promising therapeutic anti-inflammatory drug candidate. [ABSTRACT FROM AUTHOR]

tert -Butylhydroquinone (TBHQ) losses and the residual levels of 2- tert -butyl-1,4-benzoquinone (TBBQ) in tripalmitin at different heating temperatures with or without reflux over various time intervals were investigated. Heating at 120 °C resulted in the slowest TBHQ loss and the highest TBBQ levels (52.61–62.93 μg/mL). The highest TBBQ concentrations (111.73–164.67 μg/mL) at 5 and 8 h and residual concentrations of 10.23–46.95 μg/mL during heating at 170 °C over 24 h were observed. Furthermore, the potential cytotoxicity of TBBQ to RAW 264.7 cells was evaluated with the MTT assay, Hoechst 33258 staining test, and flow cytometry analysis. Results indicate that TBBQ dose- and time-dependently decreased the growth of cells and inhibited DNA synthesis by regulating the S/G2 transition. The TBBQ concentration giving 50% inhibition in RAW 264.7 cells was 10.71 μg/mL. This threshold value is lower than the residual level of TBBQ in oil, indicating the necessity for concerns over the safety of fried food in terms of TBBQ residues. [ABSTRACT FROM AUTHOR]

Oxidative stress and inflammation are important critical factors that are implicated in almost all life style disorders such as diabetes, cardiovascular disease, ulcer and cancer. Current study aimed at isolation and characterization of a furanocoumarin from Bael ( Aegle marmelos L.) fruit that can modulate both oxidative stress and inflammation effectively. Ethyl acetate extract of Bael fruit (EAFB) was subjected to HPLC for identification, purified and characterized using FTIR, NMR and ESI-MS analysis. Predominant peak of EAFB at RT 12.54 min on HPLC was identified as marmelosin with molecular weight of m/z ∼ 271.2. Marmelosin was evaluated for antioxidant, antiproliferative, apoptotic, cancer (Tyrosinase & Galectin-3) and immunomodulatory (NO, TNF-α) potentials employing standard assay systems. Marmelosin possessed potent antioxidant activity with IC 50 of ∼ 15.4 ± 0.32 μM as opposed to standard - gallic acid (IC 50 1.1 ± 0.08 μM), antiproliferative activity with IC 50 of ∼ 6.24 ± 0.16 μM as opposed to deferoxamine (∼10.8 ± 0.28 μM) and protected cells against cellular/DNA damage. Anti-inflammatory property was evident with significant reduction in the release of NO (∼3.9 fold) and TNF-α (∼3.4 fold), a pro-inflammatory cytokine, in addition to the inhibition of NFκB (∼2.7 fold), a transcription factor in Raw 264.7 cells. Marked down regulation of galectin-3 (∼5.5 folds) and tyrosinase (∼11.1 folds) by gene expression analysis substantiated by tyrosinase inhibition (IC 50 – 20.3 ± 1.26 μM Vs. Kojic acid – IC 50 – 24.1 ± 1.41 μM) and molecular docking studies strengthened the cancer modulatory property of marmelosin. In addition, marmelosin induced apoptotic bodies, chromatin condensation and nulcear blebbing in Raw 264.7 cells commending the apoptotic effect of marmelosin. Marmelosin thus displayed potential multi-potent antioxidant, anti-inflammatory and anticancer properties via TNF-α mediated Akt signaling pathway. [ABSTRACT FROM AUTHOR]

The in vivo immunomodulatory activities of Tinospora crispa have been reported but its molecular mechanisms underlying its immunomodulatory properties remains obscure and the active constituents contributing to the activities have not been identified. The present study was aimed to investigate the immunomodulatory effects of T. crispa extract (TCE) and its chemical constituents on RAW 264.7 macrophages. Six known compounds including magnoflorine and syringin were isolated by various chromatographic techniques from TCE and their structures were determined spectroscopically. A validated HPLC method was used to quantify magnoflorine and syringin in the extract. The immunomodulatory effects of TCE and its isolated compounds on chemotaxis, phagocytosis, production of inflammatory mediators including reactive oxygen species (ROS), nitric oxide (NO), prostaglandin E2 (PGE2) and pro-inflammatory cytokines which include tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 and monocyte chemoattractant protein-1 (MCP-1) on macrophages were assessed. TCE increased the chemotaxis and phagocytic activity of macrophages and significantly enhanced the production of ROS, NO and pro-inflammatory cytokines. All alkaloids isolated, specifically magnoflorine showed remarkable inducing effects on the chemotaxis, phagocytic activity, ROS and NO productions and the secretions of IL-1β, TNF-α, IL6, PGE2 and MCP-1. In contrast, syringin potently reduced the chemotaxis, phagocytic activity, ROS and NO productions and secretions of IL-1β, TNF-α, IL6, PGE2 and MCP-1. TCE showed strong immunostimulant effects on various components of the immune system and these activities were possibly contributed mainly by the alkaloids specifically magnoflorine. TCE has potential to be developed as an effective natural immunostimulant for improvement of immune-related disorders. [ABSTRACT FROM AUTHOR]

Morus bombycis Koidzumi, commonly known as silkworm mulberry, is a plant belonging to family Moraceae. It has been used in Asian countries as a traditional medicine for treating hypertension, diabetes, and inflammatory disorders. In this study, we isolated eleven compounds from the cortex of M. bombycis and evaluated their inhibitory effects on nitric oxide (NO) production as an indicator of their anti-inflammatory activities using lipopolysaccharide (LPS)-stimulated murine macrophages. Compound 4 showed the most potent inhibitory activity on NO production. It was identified as mulberrofuran K (MFK). Anti-inflammatory activity of MFK was then carried out using LPS-stimulated RAW264.7 cells. MFK suppressed the production of NO, reactive oxygen species (ROS), and proinflammatory cytokines (interleukin (IL)-1β, IL-6 and tumor necrosis factor alpha (TNF-α)) in a concentration-dependent manner. Western blot analysis revealed that MFK treatment inhibited expression levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). MFK also inhibited transcriptional activation of nuclear factor-κB (NF-κB) and extracellular-regulated kinases (ERK) 1/2 in LPS-stimulated murine macrophages. These results suggest that MFK, an anti-inflammatory constituents of M. bombycis cortex, has potential as a therapeutic candidates for preventing and treating inflammatory diseases. [ABSTRACT FROM AUTHOR]

Respiratory syncytial virus (RSV) causes acute respiratory tract infections in infants, the elderly and immunocompromised individuals. No licensed vaccine is available against RSV. We previously reported that intranasal immunization of rodents and lambs with a RSV vaccine candidate (ΔF/TriAdj) induces protective immunity with a good safety profile. ΔF/TriAdj promoted innate immune responses in respiratory mucosal tissues in vivo , by local chemokine and cytokine production, as well as infiltration and activation of immune cells including macrophages. The macrophage is an important cell type in context of both innate and adaptive immune responses against RSV. Therefore, we characterized the effects of ΔF/TriAdj on a murine macrophage cell line, RAW264.7, and bone marrow-derived macrophages (BMMs). A gene expression study of pattern recognition receptors (PRRs) revealed induction of endosomal and cytosolic receptors in RAW264.7 cells and BMMs by ΔF/TriAdj, but no up-regulation by ΔF in PBS. As a secondary response to the PRR gene expression, induction of several chemokines and pro-inflammatory cytokines, as well as up-regulation of MHC-II and co-stimulatory immune markers, was observed. To further investigate the mechanisms involved in ΔF/TriAdj-mediated secondary responses, we used relevant signal transduction pathway inhibitors. Based on inhibition studies at both transcript and protein levels, JNK, ERK1/2, CaMKII, PI3K and JAK pathways were clearly responsible for ΔF/TriAdj-mediated chemokine and pro-inflammatory cytokine responses, while the p38 and NF-κB pathways appeared to be not or minimally involved. ΔF/TriAdj induced IFN-β, which may participate in the JAK-STAT pathway to further amplify CXCL-10 production, which was strongly up-regulated. Blocking this pathway by a JAK inhibitor almost completely abrogated CXCL-10 production and caused a significant reduction in the cell surface expression of MHC-II and co-stimulatory immune markers. These data demonstrate that ΔF/TriAdj induces multiple signaling pathways in macrophages. [ABSTRACT FROM AUTHOR]

Ethnopharmocological relevance Cortex Eucommiae (CE), the bark of Eucommia ulmoides Oliv., has been traditionally used for its kidney-tonifying and bone- and tendon-enhancing properties in Korea, China, and Japan. CE has been historically prescribed for inflammatory conditions such as arthritis of the knee and ankle. Aim of the study Although CE has recently been shown to suppress inflammation in scientific studies, whether this effect involves modulation of the toll-like receptor 4 (TLR-4) pathway is currently unknown. Materials and methods The modulatory effect of CE on the TLR-4 pathway, both myeloid differentiation primary response gene 88 (Myd88)-dependent and independent, was investigated through real-time reverse transcriptase-polymerase chain reaction (RT-PCR), western blotting, and a reporter gene assay in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages. Results CE dose-dependently inhibited nitric oxide production without significant cytotoxicity with an IC 50 of 356.23 μg/mL. In addition, CE down-regulated both LPS-induced mRNA and protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) in a dose-dependent manner. CE suppressed LPS-induced activation of nuclear factor-κB (NF-κB) and the mitogen-activated protein kinase (MAPK) pathways, which together comprise the Myd88-dependent TLR-4 pathway. The phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) pathway was also down-regulated by CE in a dose-dependent manner. CE additionally suppressed LPS-induced activation of interferon-β (IFN-β) and signal transducer and activator of transcription (STAT) pathway, which is associated with the Myd88-independent TLR-4 pathway. Conclusions CE down-regulated both Myd88-dependent and independent TLR-4 pathways, thus exerting anti-inflammatory effects. These results suggest that CE may be used as a potential therapeutic agent against chronic inflammatory diseases. [ABSTRACT FROM AUTHOR]

Most fungi that grow on damp building materials produce low molecular weight compounds, some of which are known to be toxic. In this study, we tested the hypothesis that exposure to some metabolites of fungi common on damp building materials would result in time-, dose-, and compound-specific responses in the production of various chemokines by RAW 264.7 cells. Cell cultures were exposed to a 10 − 7 M or 10 − 8 M metabolite dose for 2, 4, 8 or 24 h. Metabolite concentrations used were based on those that might be expected in alveolar macrophages due to inhalation exposure from living or working in a damp building. Compared to controls, exposure provoked significant time-, dose- and compound-specific responses manifest as differentially elevated secretion of three of nine cytokines tested in culture supernatant of treated cells. The greatest number of cytokines produced in response to the metabolites tested were in andrastin A-treated cells (GM-CSF, TGFβ1, Tnf-α) followed by koninginin A (TGFβ1 and Tnf-α) and phomenone (GM-CSF, TGFβ1). Chaetoglobosin A, chaetomugilin D and walleminone exposures each resulted in significant time-specific production of Tnf-α only. This investigation adds to a body of evidence supporting the role of low molecular weight compounds from damp building materials as pathogen associated molecular patterns (PAMPs). Along with fungal glucan and chitin, these compounds contribute to the non-allergy based respiratory outcomes for people living and working in damp buildings. [ABSTRACT FROM AUTHOR]

Hentriacontane, has various pharmacological effects including anti-inflammatory, antitumor and antimicrobial activities. Its anti-inflammatory potential has been demonstrated in peritoneal macrophages. However detailed studies on other models elucidating the mechanistic description of the mode of action has not been done. Hence, the aim of the present study is to evaluate the anti-inflammatory potential of hentriacontane both in - vivo (Balb/c mice) and in-vitro (RAW 264.7 cells). Cytokine inhibition of both pro-inflammatory (TNF-α, IL-6, MCP-1 and IL-1β) and anti-inflammatory (IL-10) cytokines was studied in RAW 264.7 cells and Balb/c mice. Suppressive potential of hentriacontane on NO, PGE 2 , LTB4 and on LPS induced translocation of NF-κB in RAW 264.7 cells was studied. Further investigations on the effect of hentriacontane on phagocytic index, carrageenan induced paw oedema in mice and on organ weight were done. It was found that hentriacontane significantly reduced all the parameters of inflammation in the experiments under study at all the concentrations, 10 μM, 5 μM and 1 μM ( in-vitro ) and 5 mg/kg, 2 mg/kg and 1 mg/kg ( in-vivo ). The highest concentration used in the two models presented the most significant results. The results indicate that hentriacontane is a potent suppressor of inflammatory cytokines and other mediators. Moreover it also has regulatory effect on NF-κB. Hence, hentriacontane is a potential candidate for investigations to develop anti-inflammatory drug. [ABSTRACT FROM AUTHOR]

The heliangin is a natural agent mainly isolated from Helianthus tuberosus L. (Asteraceae). In order to investigate the anti-inflammatory effect of heliangin, several typical models in vivo and in vitro were performed. The RAW264.7 mouse macrophages cells were employed in vitro and dexamethasone were conducted as positive. The cytotoxicity results of heliangin on RAW 264.7 cells provided the safety in vitro for further study. The mRNA of TNF-α, IL-6, iNOS and COX-2 were degraded under heliangin exposure in LPS-stimulated RAW 264.7 cells. The protein expression of iNOS, COX-2 were decreased via heliangin exposure in a dose-dependent manner. Heliangin inhibited TNF-α, NO, IL-6 and PGE 2 expression levels in macrophage cells lysate. The immunocytochemistry assay showed the fluorescence image of heliangin treatment intercepted the p65 translocation process from outside to inside of nuclei triggered by LPS. Moreover, we founded that MAPK and NF-κB signaling pathway play important roles in heliangin’s activity on RAW264.7 cells. Secondly, the acute toxic study results of heliangin manifested the safety in vivo. Heliangin exerted anti-inflammation effect in a xylene-induced ear swelling in BALB/C mice and carrageenan-induced paw edema model in SD rats. The cytokines levels (TNF-α, IL-6 and PGE 2 ) were decreased. The paw tissue immunochemistry assay demonstrated the IL-6 protein level changes in carrageenan-induced paw edema model under heliangin administration. [ABSTRACT FROM AUTHOR]

Synthetic pyrethroids (SPs) are commonly used insecticides that have been detected in mammals, including humans, indicating a potential threat to human health. Bifenthrin (BF), as well as other pyrethroids, has been shown to possess neurotoxic, reproductive, hepatotoxic and nephrotoxic potential in mammals. However, studies regarding the immunotoxicity of BF and its mechanism are limited. In this study, we aim to exam the immunotoxicity of cis -BF on the murine macrophage cell line, RAW 264.7. MTT assay results demonstrated that cis -BF exposure induced apoptosis in RAW 264.7 cells in a concentration-dependent manner. We found that the expression of p53 and caspase-3 was up-regulated, while the expression of Bcl-2 was down-regulated during cis -BF-induced apoptosis. In addition, we also found that cis -BF exposure caused oxidative stress in RAW 264.7 cells in a dose-dependent manner. Interestingly, cis -BF exposure was found to inhibit the increase in transcription levels of IL-1β , IL-6 and TNF-α responding to LPS stimulation. We also found that the induced increase in IFN-β mRNA levels upon Sendai virus infection was blocked with cis -BF exposure. Finally, we found that cis -BF exposure increased ROS levels and dysregulated mRNA levels of oxidative stress-related genes in RAW 264.7 cells. The present study elucidates the immunotoxicity effect of cis -BF on macrophages and its possible underlying mechanism. The results from this study support the necessity to evaluate immune dysfunction in the risk assessment of cis -BF exposure. [ABSTRACT FROM AUTHOR]

Eight new compounds, dioscorosides G ( 1 ), H 1 ( 2 ), H 2 ( 3 ), dioscorol B ( 4 ), dioscorosides I ( 5 ), J ( 6 ), K 1 ( 7 ), and K 2 ( 8 ), together with twelve known ones ( 9 – 20 ) were obtained from the rhizomes of Dioscorea septemloba . Their structures were elucidated by chemical and spectroscopic methods. Among the known isolates, 12 – 14 , 18 , and 20 were isolated from the Dioscoreae genus for the first time. While, 9 – 11 , 15 , and 16 were firstly obtained from the plant. Moreover, all the isolates were evaluated for in vitro anti-inflammatory potential using LPS-stimulated RAW 264.7 murine macrophages, and compounds 7 , 11 , 15 , and 16 were found to display significant inhibition of nitrite production. [ABSTRACT FROM AUTHOR]

The aim of the present study was to investigate the effects of 5-bromo-2-hydroxy-4-methyl-benzaldehyde (BHMB) on inflammatory responses to lipopolysaccharide (LPS) in RAW 264.7 cells and the associated mechanism of action. BHMB concentration-dependently suppressed protein and mRNA expressions of iNOS and COX-2, thereby inhibiting the production of NO and PGE 2 in LPS-stimulated RAW 264.7 cells. BHMB also reduced the mRNA expression of TNF-α, IL-6, and IL-1β in LPS-stimulated RAW 264.7 cells. To elucidate the mechanism underlying the anti-inflammatory activity of BHMB, we investigated the effects of BHMB on the mitogen-activated protein kinase and nuclear factor-kappa B (NF-κB) pathways. BHMB suppressed the phosphorylation and degradation of IκB-α and markedly inhibited the nuclear translocation of p65 and p50 in LPS-stimulated RAW 264.7 cells. The compound also inhibited the LPS-stimulated phosphorylation of ERK and p38. Taken together, these results illustrated that BHMB suppresses pro-inflammatory mediator and cytokine expression in LPS-stimulated RAW 264.7 cells by inhibiting the phosphorylation of ERK and p38 and the activation of NF-κB. [ABSTRACT FROM AUTHOR]

Protein interacting with C kinase 1 (PICK1) is a scaffolding protein mainly implicated in neurological diseases, however, the function of PICK1 in acute liver injury (ALI) remains unknown. Our study found a dramatical decrease in mRNA and protein levels of PICK1 in liver tissues and isolated Kupffer cells (KCs) from the liver in mice with ALI. Furthermore, pretreatment the mice with ALI with FSC-231, a pharmacological inhibitor of PICK1, could significantly augment inflammatory response. Furthermore, in vitro studies showed that both lipopolysaccharide (LPS) and interferon gamma (IFN-γ) significantly reduced the expression of PICK1, while IL-4 elevated its expression in RAW 264.7 cells. Additionally, over-expression of PICK1 inhibited the expression of M1 biomarkers by suppressing NF-κB activity, and enhanced the expression of M2 biomarkers by promoting STAT6 activity. In contrast, knockdown of PICK1 or FSC-231 pretreatment promoted M1 polarization and suppressed M2 polarization. Besides, caveolin-1 was identified as a potential target gene controlled by PICK1 in RAW 264.7 cells. Mechanistic investigation revealed a dual role of PICK1 in regulating macrophage polarization and implied PICK1 as a potential therapeutic target in ALI. [ABSTRACT FROM AUTHOR]

Kaempferol 3- O -(2G-glucosylrutinoside)-7- O -glucoside (KGG) has isolated from Hosta plantaginea flowers and possessed an inhibitory effect on cyclooxygenase 2 (COX-2), could be effective in inhibiting inflammation. However, the anti-inflammatory activity and mechanism of KGG remain unknown. In this study, for the first time, the anti-inflammatory effect of KGG and its potential molecular mechanisms were explored in cells. KGG had no cytotoxicity at concentrations of 1.25, 2.5, 5, 10, 20, and 40 μM by Cell Counting kit-8 assay in RAW 264.7 cells. Besides, KGG concentration-dependently (1.25, 2.5, and 5 μM) inhibited secretions of nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and IL-6 in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Western blot showed that the phosphorylation of nuclear factor kappa-B (NF-κB) p65, inhibitor of NF-κB (IκB), p38 MAPK, c-Jun N-terminal kinases (JNK), extracellular signal-regulated kinase (Erk), and protein kinase B (Akt), together with inducible nitric oxide synthase (iNOS) and COX-2 were significantly attenuated by KGG (1.25, 2.5, and 5 μM) in a concentration-dependent relationship. Meanwhile, KGG remarkably enhanced the protein expression of IκB. Taken together, KGG may be one of bioactive phytochemicals from H. plantaginea flowers, and be an anti-inflammatory agent via inhibiting NF-κB, mitogen-activated protein kinases (MAPKs), and Akt signaling pathways. [Display omitted] • Kaempferol 3- O -(2G-glucosylrutinoside)-7- O -glucoside (KGG) exerts anti-inflammatory effect through blocking NF-κB, MAPKS, and Akt pathways. • KGG might be a promising candidate against inflammation. [ABSTRACT FROM AUTHOR]

[Display omitted] • DS notably attenuated the inflammatory response through inhibition the NF-κB signaling pathway in LPS-induced RAW 264.7 cells in vitro. • DS markedly suppressed NF-κB and Wnt/β-Catenin signaling pathways in DSS-induced colitis mice in vivo. • DS could be further developed as a promising drug alternative for the treatment of ulcerative colitis. Daurisoline (DS) is one of the most abundant alkaloids extracted from the rhizome of Menispermum Dauricum DC, which is traditionally used to treat inflammatory diseases, especially intestinal inflammation. In this study, we established lipopolysaccharide (LPS)-induced RAW 264.7 macrophages in vitro and Dextran sulfate sodium (DSS)-induced colitis mice model in vivo to investigate the anti-inflammatory effect of DS and its underlying mechanisms. Disease activity index (DAI) was detected during drug intervention. The colon length, macroscopic changes and histopathological scores were adopted to observe the physiological status and the colon injury. The apoptosis of intestinal mucosa was detected using TUNEL. In addition, involved molecular indicators were measured by ELISA kits, RT-qPCR, immunofluorescence (IF), immunohistochemistry (IHC) and western blotting. The vitro experiments indicated that DS significantly suppressed the production of Nitric oxide (NO), reactive oxygen species (ROS) and glutathione (GSH), as well as inhibited the expression of NF-κB signaling pathway in RAW 264.7 cells induced by LPS. Consistent with the vitro experimental results, different doses of DS significantly reduced the incidence of diarrhea, DAI, shortening of the colon, visible damage and histological damage in DSS-induced colitis mice. Moreover, DS treatment decreased the levels of pro-inflammatory mediators cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE2) and interleukin (IL)-1β, and increased the anti-inflammatory cytokines IL-4 and IL-10 in colon tissues. RT-qPCR, western blotting and immunofluorescence analyses further demonstrated that DS inhibits the expression of Wnt/β-Catenin pathway. We reported for the first time that DS may be an active ingredient in treating ulcerative colitis. Its mechanism might be related to the regulation of the NF-κB and Wnt/β-Catenin signaling pathway. [ABSTRACT FROM AUTHOR]

Owing to the serious adverse effects caused by pyrimethamine and sulfadiazine, the drugs commonly used to treat toxoplasmosis, there is a need for treatment alternatives for this disease. Nanotechnology has enabled significant advances toward this goal. This study was conducted to evaluate the activity of biogenic silver nanoparticles (AgNp-Bio) in RAW 264.7 murine macrophages infected with the RH strain of Toxoplasma gondii. The macrophages were infected with T. gondii tachyzoites and then treated with various concentrations of AgNp-Bio. The cells were evaluated by microscopy, and culture supernatants were collected for ELISA determination of their cytokine concentration. Treatment with 6 μM AgNp-Bio reduced the infection and parasite load in infected RAW 264.7 macrophages without being toxic to the cells. The treatment also induced the synthesis of reactive oxygen species and tumor necrosis factor-alpha (both pro-inflammatory mediators), which resulted in ultrastructural changes in the tachyzoites and their intramacrophagic destruction. Our findings suggest that AgNp-Bio affect T. gondii tachyzoites by activating microbicidal and pro-inflammatory mechanisms and may be a potential alternative treatment for toxoplasmosis. [ABSTRACT FROM AUTHOR]

Geraniin, a major polyphenolic compound of Geranium sibiricum L, has long been used as an important Chinese herbal medicine for the treatment of a variety of inflammatory pathologies. However, the underlying anti-inflammatory molecular mechanisms of this compound are not clear. The aim of the present study was to investigate the anti-inflammatory activities of geraniin and elucidate the underlying mechanisms. The anti-inflammatory effects of geraniin were studied by using lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Geraniin suppressed the inducible nitric oxide synthase (iNOS) expression, and inhibited reactive oxygen species (ROS) production. Subsequent studies demonstrated that geraniin effectively reduced production of NO and pro-inflammatory cytokines. These effects were mediated by impaired translocation of nuclear factor (NF)-κB and inhibition of the phosphorylation of Akt in LPS-stimulated RAW 264.7 cells. Furthermore, geraniin induced heme oxygenase-1 (HO-1) expression via activation of transcription factor Nrf2. This study gives scientific evidence that geraniin inhibits the LPS-induced expression of inflammatory mediators via suppression of Akt-mediated NF-κB pathway as well as up-regulation of Nrf2/HO-1 pathway, indicating that geraniin has a potential application in inflammatory conditions. [ABSTRACT FROM AUTHOR]

Six new cyclopentenyl esters, avellaneine A–F ( 1 – 4, 7 , 8 ), two new cyclopentyl esters, avellaneine G, H ( 9 , 10 ), along with two known cyclopentenyl esters were obtained from water extract of the inner bark of Tabebuia avellanedae Lorentz ex Griseb. The chemical structures of the new compounds were elucidated by spectroscopic techniques. The anti-inflammatory effects of these compounds were determined on LPS-stimulated RAW 264.7 cell line. Some of the tested compounds ( 2 , 3 , 4 , 6 , 7 ) reduced the NO production in a dose-dependent manner, while 6 and 7 decreased PGE 2 production in a dose-dependent manner, without altering cell viability. Data presented in this research indicated that Tabebuia avellanedae 's ethnopharmacological action of treating inflammatory diseases was based on the constituents which exert a significant anti-inflammatory effect on inflammatory responses. [ABSTRACT FROM AUTHOR]

Lactobacillus species are potential probiotic bacteria for humans because of their capacity to improve certain biological functions in the host's immune system. In this study, we focused on three peptidoglycans (PGNs) derived from different Lactobacillus strains and investigated each PGN's anti-inflammatory capacity. Each PGN was analyzed using HPLC, MALDI–TOF/TOF MS and FTIR. All three PGNs displayed a β-1,4-linked N -acetylmuramic acid (MurNAc) and N -acetylglucosamine (GlcNAc) structure with some modifications in the polypeptides at the end of the MurNAc residue. In a new insight, we found that PGNs inhibit the release of inflammatory cytokines in LPS-induced RAW 264.7 cells; a capacity that may be related to the TLR-4 pathway. The goal for exploring PGN diversity in Lactobacillus strains is to better understand the potential use of Lactobacillus PGNs in food and pharmaceutical applications. [ABSTRACT FROM AUTHOR]

Asthma is a heterogeneous airway inflammatory disease, which is associated with Th2 cytokine-driven inflammation and non-Th2, TNF-α mediated inflammation. Unlike Th2 mediated inflammation, TNF-α mediated asthma inflammation is generally insensitive to inhaled corticosteroids (ICS). ASHMITM, aqueous extract of three medicinal herbs— Ganoderma lucidum ( G. lucidum ), Sophora flavescens Ait ( S. flavescens ) and Glycyrrhiza uralensis Fischer ( G. uralensis ), showed a high safety profile and was clinically beneficial in asthma patients. It also suppresses both Th2 and TNF-α associated inflammation in murine asthma models. We previously determined that G. uralensis flavonoids are the key active compounds responsible for ASHMITM suppression of Th2 mediated inflammation. Until now, there are limited studies on anti-TNF-α compounds presented in ASHMITM. The objective of this study was to isolate and identify TNF-α inhibitory compounds in ASHMITM. Here we report that G. lucidum , but not the other two herbal extracts, S. flavescens or G. uralensis inhibited TNF-α production by murine macrophages; and that the methylene chloride (MC)–triterpenoid-enriched fraction, but not the polysaccharide-enriched fraction, contained the inhibitory compounds. Of the 15 triterpenoids isolated from the MC fraction, only ganoderic acid C1 (GAC1) significantly reduced TNF-α production by murine macrophages (RAW 264.7 cells) and peripheral blood mononuclear cells (PBMCs) from asthma patients. Inhibition was associated with down-regulation of NF-κB expression, and partial suppression of MAPK and AP-1 signaling pathways. Ganoderic acid C1 may have potential for treating TNF-α mediated inflammation in asthma and other inflammatory diseases. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Orostachys japonicus A. Berger (Crassulaceae) (OJ), well-known as Wa-song in Korea is a medicinal plant with immunoregulatory, anti-febrile, antidote, and anti-cancer activities. This study was aimed at evaluating the immunostimulatory effect of O. japonicus A. Berger and its possible mechanisms of action. Materials and methods To evaluate the effect of OJ aqueous extract on macrophage activity, we evaluated the modulation of macrophage activation state by observing structural (phagocytic activities) and the production of nitric oxide increase. The effect of OJ aqueous extract on RAW264.7 cell viability were assessed using Cell Counting Kit (CCK)-8 assay. HPLC analysis was performed to identify potential active compounds of this extract. Results The biological investigations indicated that OJ aqueous extract, among others, possessed the highest macrophage activation as indicated by NO production yield. The results showed that OJ aqueous extract exhibited antioxidant effects, which included scavenging activities against DPPH radicals. OJ aqueous extract increased the phagocytic activity of RAW 264.7 cells against IgG-opsonized red blood cells (RBC). The level of phosphorylated Syk kinase was increased in OJ aqueous extract-treated group as compared to control. Phosphorylation of PLC-γ was increased in the OJ aqueous extract-treated groups. Quercetin-3- O -rhamnose and kaempferol-3- O -rhamnose was detected in OJ aqueous extract by HPLC analysis. Conclusions OJ aqueous extract might play a pivotal ethnopharmacologic role as an immunostimulatory agent by promoting Fc gamma receptor (FcγR)-mediated phagocytosis of IgG-opsonized RBCs. On the basis of our results, OJ aqueous extract can enhance innate immunity and may serve as an adjuvant for tumor treatment. [ABSTRACT FROM AUTHOR]

A new dual chemosensor ( Z )-2-(anthracen-9-ylmethylene)- N -phenylhydrazinecarbothioamide R 1 was synthesized by incorporating a thiourea derivative as a binding unit and an anthracene group as a fluorescence signalling unit. R 1 selectively binds with Cu 2+ and Hg 2+ ions in aqueous medium. R 1 exhibits highly selective recognition towards F − ion and picric acid in organic medium. Fluorescence quenching was observed in case of R 1 with Cu 2+ and Hg 2+ ions over commonly coexistent metal ions. Furthermore, fluorescence imaging experiments of Cu 2+ and Hg 2+ ions in living RAW 264.7 cells demonstrate its value of practical applications in biological systems. [ABSTRACT FROM AUTHOR]

CDDO-Me, initiated in a phase II clinical trial, is a potential useful therapeutic agent for cancer and inflammatory dysfunctions, whereas the therapeutic efficacy of CDDO-Me on LPS-induced acute lung injury (ALI) has not been reported as yet. The purpose of the present study was to explore the protective effect of CDDO-Me on LPS-induced ALI in mice and to investigate its possible mechanism. BalB/c mice received CDDO-Me (0.5 mg/kg, 2 mg/kg) or dexamethasone (5 mg/kg) intraperitoneally 1 h before LPS stimulation and were sacrificed 6 h later. W/D ratio, lung MPO activity, number of total cells and neutrophils, pulmonary histopathology, IL-6, IL-1β, and TNF-α in the BALF were assessed. Furthermore, we estimated iNOS, IL-6, IL-1β, and TNF-α mRNA expression and NO production as well as the activation of the three main MAPKs, AkT, IκB-α and p65. Pretreatment with CDDO-Me significantly ameliorated W/D ratio, lung MPO activity, inflammatory cell infiltration, and inflammatory cytokine production in BALF from the in vivo study. Additionally, CDDO-Me had beneficial effects on the intervention for pathogenesis process at molecular, protein and transcriptional levels in vitro . These analytical results provided evidence that CDDO-Me could be a potential therapeutic candidate for treating LPS-induced ALI. [ABSTRACT FROM AUTHOR]

Six new phenolics, scutellariosides A–F ( 1 – 3 , 5 – 6 , and 8 ), together with six known compounds ( 4 , 7 , 9 – 12 ) were isolated from the whole plant of Scutellaria indica (Labiatae). The chemical structures of these compounds were determined by spectroscopic analyses including 2D NMR. Their anti-inflammatory activities were evaluated against LPS-induced NO production in macrophage RAW 264.7 cells. Among them, compounds 10 – 12 had inhibitory effects with IC 50 values ranging from 7.2 to 27.8 μM. Compound 12 reduced LPS-induced iNOS expression in a dose-dependent manner. [ABSTRACT FROM AUTHOR]

The tandem mass spectrometry, ion mobility, and normal phase HPLC of isomeric phosphatidylglycerol (PG) and bis(monoacylglycerol) phosphate (BMP) have been investigated in this study with the objective of differentiating these unique classes of lipids. Measurement of ion mobility using the traveling wave method for negative molecular and product ions from isomeric PG and BMP yielded identical results, but different ion mobilities were observed for positive product ions arising from collision-induced dissociation (CID). The fastest moving positive product ions from the ion mobility analysis of BMP(18:1/18:1) were monoglyceride-like, and the slowest moving product ions from this BMP corresponded to [M + H − 2H 2 O] + , which were readily observed for BMP but were only at very low abundance in the CID spectra of PG. The major product ions observed from the sodium adduct of PG(18:1/18:1) were consistent with diglyceride-like ion formation, but for BMP(18:1/18:1) only monoglyceride-like product ions were formed. The usefulness of ion mobility separation was tested with the selection of positive product ions derived from the isomeric PG and BMP molecular species in the lipid extract of RAW 264.7 cells. The ion mobility spectra of monoglyceride-like ions derived from BMP species with various esterified fatty acyl groups displayed some separation in ion mobility based on fatty acyl chain length and presence of a double bond in the acyl chain. The mechanism of ion formation of the diglyceride- and monoglyceride-like ions from PG and BMP respectively was examined using deuterium-labeled species including PG(D 31 16:0/18:1) and PG and BMP labeled by deuterium exchange. [ABSTRACT FROM AUTHOR]

Here we aimed to investigate the anti-inflammatory effects of calcium citrate in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The anti-inflammatory effects of calcium citrate were investigated by assessing pro-inflammatory factors (NO, ROS, NF-κB, iNOS, and COX-2) and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α). Treatment of cells with calcium citrate (10–100 μM) significantly reduced the generation of intracellular reactive oxygen species and increased the activities of the antioxidant enzymes superoxide dismutase, catalase, and glutathione peroxidase in LPS-stimulated macrophages. Calcium citrate was further shown to inhibit NO production in LPS-stimulated RAW 264.7 cells. The expression levels of iNOS, COX-2, and NF-κB were also suppressed by treatment with calcium citrate. Calcium citrate was furthermore found to significantly inhibit the production of IL-1β, IL-6, and TNF-α in response to LPS-stimulation. These findings demonstrate that calcium citrate may be an effective anti-inflammatory agent. [ABSTRACT FROM AUTHOR]

The inflammatory response protects our body from bacteria and tumors, but chronic inflammation driven by the persistent activation of macrophages can lead to serious adverse effects including gastrointestinal problems, cardiac disorders, and a sore throat. Part of the ongoing research is focused on searching for antiinflammatory compounds from natural sources, so we investigated the effects of hardy kiwis ( Actinidia arguta , Lauraceae) stems on inflammation induced by lipopolysaccharide (LPS) in Raw 264.7 cells to test the hypothesis that antiinflammatory effects of A. arguta stems were exerted through the inhibition of the nuclear factor (NF)- κ B pathway. The methanol extract of A. arguta (20 μ g/mL) stems lowered nitric oxide production in LPS-stimulated Raw 264.7 cells by 40%. It was then partitioned with hexane, chloroform, ethyl acetate, butanol, and water based on the polarity of each compound. Among the 5 layers, the chloroform layer had the greatest inhibitory effect on LPS-stimulated nitric oxide production and inducible nitric oxide synthase mRNA expression in Raw 264.7 cells. However, the levels of prostaglandin E2 and cyclooxygease 2 were not altered. On the other hand, treatment of cells with the chloroform layer of A. arguta before LPS stimulation also reduced the mRNA expression of proinflammatory cytokines including tumor necrosis factor α and interleukin 1 β . Nuclear translocation of NF- κ B p50 and p65 subunits induced by LPS was also inhibited by treatment with the chloroform layer of A. arguta . This was accompanied with the reduced phosphorylation of mitogen-activated protein kinases including extracellular signal-regulated protein kinase 1/2, c-Jun N-terminal protein kinase, and p38. Taken together, these results suggest that chloroform layer of A. arguta exerted antiinflammatory effects by the inhibition of mitogen-activated protein kinase phosphorylation and nuclear translocation of NF- κ B. [ABSTRACT FROM AUTHOR]

[Display omitted] • Surface treatment can enhance the biocompatibility of the 3D printed Ti scaffolds. • Etched Ti5Cu scaffolds showed higher proliferation, lowest apoptosis rate and the best early osteogenic ability. • Higher concentration powders inhibited the proliferation and osteogenic differentiation of MC3T3-E1 cells, as well as promoted apoptosis. 3D printed medical titanium alloy has shown great advantages in manufacturing complex and personalized scaffolds for tissue engineering. To promote the repair of bone defects, fabrication of a scaffold with high accuracy and excellent cell behaviors is required. However, the partially melted or un-melted powders must be removed in the post-process. In this study, the effects of unfused metal powders on the biocompatibility of 3D printed Ti alloy scaffolds were analyzed by co-culture with MC3T3-E1 cells and RAW 264.7 cells, and the surface treatment strategy was proposed. Results showed that surface treatment can enhance the biocompatibility of the 3D printed Ti alloy scaffolds. The vitro tests with MC3T3-E1 cells demonstrated that the ETi5Cu (etched Ti5Cu) showed the better cell proliferation and osteogenic potential, and the lower cell apoptosis rates. Moreover, higher concentration of powders showed inhibitory effects on cytocompatibility and osteogenesis of MC3T3-E1 cells, and promoted the polarization of macrophages to M1 type. In summary, the removal of metal powders on the 3D printed scaffolds plays an important role in the biological performance, which provided a promising strategy for the application of Ti implants. [ABSTRACT FROM AUTHOR]

Abstract: Three sulfated phenolic compounds, juglanin B (11R)-O-sulfate (1), myricetin 3´-O-sulfate (2), and ampelopsin 3´-O-sulfate (3), were isolated from the leaves of Myrica rubra. Compound 1 was a new sulfated lignan, 2 was a new sulfated flavone, and 3 was a known sulfated flavone. The structures of the new compounds (1 and 2) were determined by acid hydrolysis and spectroscopic methods, including IR, FAB-MS, 1D and 2D NMR. The inhibitory activities of compounds 1–3 and their hydrolysates (1a–3a) against LPS-induced cytokine (TNF-α, IL-1β, and IL-6) production in macrophage RAW 264.7 cells were evaluated. The 2 new compounds (1 and 2) and their aglycones (1a and 2a) significantly reduced LPS-induced expression of iNOS and COX-2 proteins. [Copyright &y& Elsevier]

Abstract: BaeR is the response regulator of the two-component system, BaeSR, found in Escherichia coli (E. coli) and Salmonella. Several biological functions of BaeR, related to multidrug efflux and bacterial virulence, have been described. Herein, we report a putative function of BaeR during inflammatory response of the host by using BaeR protein of Salmonella enterica Paratyphi A (S. Paratyphi A) origin overexpressed in E. coli, and RAW 264.7 and THP-1 cells as in vitro models. BaeR (3 μg/ml) upregulated iNOS mRNA expression in both cell lines, and induced significant production of NO. Greater than ten-fold (TNF-α), 24-fold (IL-1β) and 156-fold (IL-6) increases in mRNA expression levels were observed in THP-1 cells treated with BaeR, compared to untreated controls. Furthermore, an eight-fold (IL-1β), 12-fold (IL-6) and 41-fold (TNF-α) higher protein concentrations were observed in RAW 264.7 cells stimulated with BaeR, compared to control cells. Immunoblot analysis showed BaeR-induced phosphorylation of the MAPKs (ERK 1/2, JNK and p38 MAPK) in RAW 264.7 cells. Pharmacological inhibition of the three MAPKs using specific inhibitors resulted in significant reduction of BaeR-induced NO production and iNOS mRNA expression by inhibitors of JNK and p38 MAPK. Also, all inhibitors of the MAPKs significantly attenuated BaeR-induced IL-1β, IL-6 and TNF-α at both transcript and protein levels with different degrees of inhibition. Taken together, our data suggest that BaeR is a putative inducer of inflammatory response and the MAPKs are involved in the process. [Copyright &y& Elsevier]

Many natural resources obtained from plants have been studied for their utility as host defense potentiators. In the present study, we investigated whether a glycoprotein fraction from rice (Oryza sativa) bran (GFRB) could modulate immune responses such as the production of nitric oxide (NO), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and IL-10 in the RAW 264.7 macrophage cell line. GFRB, which contained 65.7% of protein and 7.7% of total sugar, was prepared by treating an aqueous extract of rice bran with 80% (NH4)2SO4 and the extraction yield was 4.9%. GFRB consisted of 5 bands with varying molecular weights by SDS-PAGE and remarkably improved production of NO in RAW 264.7 murine macrophage cells, up to approximately 10-fold compared to the normal control at 100μg/mL concentration. In RAW 264.7 cells treated with 50μg/mL GFRB, released levels of various cytokines such as TNF-α, IL-1β, IL-6, and IL-10 were 2824.4±90.7, 224.5±4.0, 524.3±4.8, and 143.0±9.5pg/mL, respectively, which were higher than the levels in normal controls. Moreover, GFRB exhibited no cytotoxicity. According to the results of region-selective enzyme hydrolysis, the immune responses against GFRB were elicited by the glycans in the GFRB. These results show the potential of GFRB as a functional therapeutic agent with demonstrable immunostimulatory activity. [ABSTRACT FROM AUTHOR]

Abstract: Numerous studies have shown various relationships between foods with a high nutritional value and a robust immune response, particularly studies that have focused on host protection and cytokine networks. This study aimed to clarify the role played by the procyanidin trimer C1 in innate and adaptive immunity. Procyanidin C1 did not exert cytotoxicity at concentrations ranging from 7.8 to 62.5μg/ml in macrophage cells; therefore, concentration of 62.5μg/ml was used as the maximum dose of procyanidin C1 throughout subsequent experiments. Procyanidin C1 enhanced inducible nitric oxide synthase-mediated nitric oxide production in a concentration-dependent manner. In addition, procyanidin C1 functionally induced macrophage activation by augmenting the expression of cell surface molecules (CD80, CD86, and MHC II) and proinflammatory cytokine production (tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6) via activation of mitogen-activated protein kinase (MAPK), e.g., p38, ERK, and JNK and nuclear factor (NF)-κB signaling pathways. Interestingly, procyanidin C1 effectively polarized T helper type 1 (Th1) by secreting Th1-mediated cytokines (interferon-γ, IL-12p70, and IL-2) and inducing splenocyte proliferation, indicating that procyanidin C1 contributes to Th1 polarization of the immune response. Accordingly, these findings confirms that the procyanidin C1 induces macrophage activation via NF-κB and MAPK pathways, leading to Th1 polarization in murine splenocytes, which suggests that procyanidin C1 regulates innate and adaptive immunity by macrophage activation and Th1 polarization. [Copyright &y& Elsevier]

The young branches of C. cassia Blume (Cinnamomi Ramulus; Guizhi; 桂枝; C. cassia twigs) have long been used as an anti-pyretic, anti-rheumatic, anti-spasmodic and stomachic in traditional medicine. The aim of this study was to test the anti-inflammatory, anti-nociceptive, and anti-arthritic effects of Cinnamomum cassia twigs in acute and chronic arthritis rats. Subcutaneous injection of carrageenan for acute inflammation and complete Freund's adjuvant (CFA) for chronic arthritis was carried out in the hind paw of SD rats. The paw volume was measured by a plethysmometer; thermal hyperalgesia was tested using a thermal plantar tester; hyperalgesia was evaluated by ankle flexion evoked vocalizations. The c-Fos expression in the lumbar spinal cord was measured with the avidin-biotin-peroxidase technique. The nitric oxide (NO) generation in lipopolysaccharide (LPS)-induced RAW 264.7 cells was tested by Griess assay. An 80% ethanoic extract of the C. cassia twigs exhibited chronic anti-inflammatory and anti-arthritic activities by reducing the edema volume in the paws of CFA-induced chronic arthritis in rats. In addition, it showed analgesic effects through the recovery of the paw withdrawal latency stimulated by thermal hyperalgesia, and suppressing the vocalization scores evoked by ankle flexion in the hind paws of the arthritis rats. It also controlled c-Fos expression in the lumbar spinal cord of the arthritis rats. Moreover, the addition its 80%-ethanoic extract, specifically, its ethyl acetate fraction, powerfully suppressed the paw swelling in carrageenan-stimulated arthritis and the NO production in LPS-induced murine immune cells. C. cassia twigs may act as a viably sufficient therapeutic or preventive candidate for osteoarthritis and rheumatoid arthritis; additionally, it could prevent gastrointestinal damage with its gastric protection. [Display omitted] [ABSTRACT FROM AUTHOR]

Abstract: Four new lanostane triterpenes, butyl lucidenate P (1), butyl lucidenate D2 (2), butyl lucidenate E2 (3) and butyl lucidenate Q (4) along with 11 known compounds (5–15) were isolated from the fruiting bodies of Ganoderma lucidum. Their chemical structures were established mainly by 1D and 2D NMR techniques and mass spectrometry. Their anti-inflammatory activity was evaluated against LPS-induced NO production in macrophage RAW 264.7 cells. Compounds 1, 3, 4, 9, 10 and 15 showed inhibitory potency with IC50 values of 7.4, 6.4, 4.3, 9.4, 9.2 and 4.5μM, respectively. Compounds 1, 3 and 15 dose-dependently reduced the LPS-induced iNOS expressions. Preincubation of cell with 1, 3 and 15 significantly suppressed LPS-induced expression of COX-2 protein. [Copyright &y& Elsevier]

Abstract: Aims of the study: We recently reported the potential antioxidant and anti-inflammatory activities of umbelliferone 6-carboxylic acid (UMC) isolated from the whole plants of Angelica decursiva. In this study, we elucidated the anti-inflammatory mechanisms of UMC in vitro and in vivo. Methods: The inhibitory effects of UMC on the production of nitric oxide (NO), prostaglandin E2 (PGE2), and tumor necrosis factor-α (TNF-α), the expression of nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), the activation of nuclear factor kappa B (NF-κB) were evaluated using lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. The reactive oxygen species (ROS) generation inhibitory activity of UMC was evaluated using t-butyl hydroperoxide (t-BHP)-induced RAW 264.7 cells. Furthermore, the in vivo anti-inflammatory activity of UMC was evaluated using carrageenan induced mouse paw edema model. Results: UMC dose-dependently inhibited NO and PGE2 production by down-regulating iNOS and COX-2 protein expression in LPS-stimulated RAW 264.7 macrophages. UMC also suppressed the production of the proinflammatory cytokine TNF-α in LPS stimulated RAW 264.7 cells in a concentration dependent manner. In addition, UMC dose-dependently prevented LPS-induced nuclear translocation of NF-κB in RAW 264.7 macrophages. Furthermore, UMC exhibited the inhibitory activity against t-BHP-induced ROS generation in RAW 264.7 cells with an IC50 value of 705.1μg/ml. Moreover, UMC inhibited λ-carrageenan induced mouse paw edema by 70.40 and 60.20% at doses of 50 and 25mg/kg body weight, respectively. Conclusion: The combined results of this study indicate that UMC is an important anti-inflammatory constituent of A. decursiva and its anti-inflammatory effect was due to its ability to inhibit the production of inflammatory mediators via inhibition of NF-κB activation pathway. [Copyright &y& Elsevier]

Abstract: We used the modified Mannich condensation to synthesize three closely-related series of 7-alkylated 3-ABNs 1–5 viz., 7-methylated 2,4-diaryl-3-azabicyclo[3.3.1]nonan-9-ones (7-Me ABNs 1–5), 7-ethylated 2,4-diaryl-3-azabicyclo[3.3.1]nonan-9-ones (7-Et ABNs 1–5) and 7-tert-pentylated 2,4-diaryl-3-azabicyclo[3.3.1]nonan-9-ones (7-tert-pentyl ABNs 1–5). All compounds yielded good as single isomers by the use of PPA·SiO2 as a heterogeneous Bronsted acidic catalyst. The 1D, 2D NMR, and single-crystal XRD interpretations unambiguously characterized the stereochemistry of the synthesized compounds. In solution as well as solid-state, all compounds exist in the twin-chair conformation with equatorial orientations of all substitutions, despite their nature and positions. The chemical methods viz., DPPH, reducing power, and phospho-molybdenum methods identified some of the target curcumin analogs as active compounds. Among them, 7-Me ABN 4 (7-methyl-2,4-bis(3-methoxy-4-hydroxyphenyl)-3-azabicyclo[3.3.1]nonan-9)-one exerted the best antioxidant profile that comparable to standard l-ascorbic acid, α-tocopherol and curcumin. Hence, we evaluated further for its intracellular ROS inhibition potency on RAW 264.7 macrophage cells, and found to be effective as well as non-toxic at 100μM. [Copyright &y& Elsevier]

Abstract: We have demonstrated previously that the acid-treated multi-walled carbon nanotubes (aci-MWCNTs) and taurine functionalized MWCNTs (tau-MWCNTs) induced differential pulmonary toxicity in mice after instillation exposure. In order to compare differences of cytotoxicity between the aci- and tau-MWCNTs, RAW 264.7 cells (a murine macrophage cell line) were chosen to be exposed to the aci- and tau-MWCNTs at concentrations of 0, 5, 20, 40, and 80μg/ml for 12 or 24h respectively. The results showed that although the aci- and tau-MWCNTs induced only mild decrease in cell viability to RAW 264.7 cells, the two types of MWCNTs elicited significant increase in apoptosis and decreased ability in cellular phagocytosis. Moreover, by using the specific inhibitors, we found that the scavenger receptors (SR) and caspase-9 were actively involved in the apoptosis induced by the aci- and tau-MWCNTs. The taurine functionalized MWCNTs (tau-MWCNTs) showed less cytotoxicity and apoptotic effect to RAW 264.7 cells than those of aci-MWCNTs. Taken together, the results indicated the important role of scavenger receptors and mitochondria in the apoptosis induced by MWCNTs. [Copyright &y& Elsevier]

Abstract: Suppression of interleukin (IL)-6 production has beneficial effects against various inflammatory diseases. Through a rapid screening system, we found that nitazoxanide, or 2-acetyloxy-N-(5-nitro-2-thiazolyl) benzamide, which is a well-known antiparasitic agent, suppressed lipopolysaccharide (LPS)-induced production of IL-6 from RAW 264.7 cells and mouse peritoneal macrophages, with 50% inhibitory concentrations (IC50s) of 1.54mM and 0.17mM, respectively. Nitazoxanide also inhibited the LPS-induced expression of IL-6 mRNA in RAW 264.7 cells. To investigate the effects of nitazoxanide in vivo, we orally administered nitazoxanide at a dose of 100mg/kg to mice 2h before a 1-mL intraperitoneal injection of 4% thioglycollate (TG). Six hours after TG injection, plasma IL-6 levels were markedly lower (by 90%) than the levels in vehicle-treated mice. These data suggest that nitazoxanide could be a promising lead compound for agents against various diseases associated with overproduction of IL-6. [Copyright &y& Elsevier]

Abstract: The aim of this study is to investigate the inhibitory effects of aminoethyl-chitooligosaccharides (AE-COS) on oxidative stress in mouse macrophages (RAW 264.7 cells). The inhibitory effects of AE-COS on DNA and protein oxidation were studied in RAW 264.7 cells. Furthermore, free radical scavenging effect of AE-COS were determined in RAW264.7 cells by 2′,7′-dichlorofluorescein (DCF) intensity and intracellular glutathione (GSH) level. AE-COS also inhibited myeloperoxidase (MPO) activity in human myeloid cells (HL-60). These results suggest that AE-COS acts as a potential free radical scavenger in RAW 264.7 cells. [Copyright &y& Elsevier]

Abstract: Ethnopharmacological relevance: Orostachys japonicus A. Berger (O. japonicus) is known to reduce the risk of many diseases. Aim of the study: We investigated the anti-inflammatory effects of the dichloromethane (DCM) fraction from O. japonicus (OJD) in LPS-stimulated RAW 264.7 cells. Materials and methods: NO was measured using the Griess method. Key pro-inflammatory cytokines and mediators including IL-1β, TLR4, iNOS, and COX-2; 2 important pro-inflammatory transcription factors, NF-κB p65 and IκBα; and MAPKs such as ERK1/2, JNK, and p38 were analyzed by Western blotting. Results: OJD significantly inhibited NO production, IL-1β, TLR4, iNOS, and COX-2 expression in LPS-stimulated cells. Additionally, it inhibited LPS-induced NF-κB p65 activation via inhibition of IκBα phosphorylation. Furthermore, phosphorylation of p38 and JNK was suppressed by OJD in a dose-dependent manner in the LPS-stimulated RAW 264.7 cells. Conclusions: Our data suggest that OJD inhibits the inflammatory response via suppression of NF-κB activation and MAPK signaling. [Copyright &y& Elsevier]