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Four new hydroperoxyfurancembranoids, briaviodiols B–E (1–4), were obtained from an octocoral identified as Briareum violaceum. The structures of cembranoids 1–4 were established by using spectroscopic methods and comparing the spectroscopic data with those of known related analogues. Compounds 1, 3, and 4 were found to exhibit significant in vitro anti-inflammatory activity in LPS-induced RAW 264.7 macrophage cells by inhibiting the expression of iNOS protein.

Chaga mushroom, Inonotus obliquus, was used as food and nutrient food and traditional herbs in Russia, China and Japan, with anti-inflammatory and anticancer activities. Chemical investigations of the fruiting bodies of Chaga were carried to uncover the bioactive metabolites. As a result, seven undescribed lanostane-type triterpenoids, namely inonotusols H–N, were isolated, and all lanostanoids remarkably inhibited NO production in lipopolysaccharide-stimulated BV-2 microglial cells. Of these, inonotusols I and L presented the most potent inhibitory effects on inducible nitric oxide synthase (iNOS) and NO production without any significant cytotoxicity. Molecular docking studies confirmed the capacity of inonotusols I and L to interact with iNOS protein. Structure-activity relationships were also discussed. These results indicated that the potential anti-inflammatory effects of inonotusols I and L in microglial BV-2 cells may be imparted through suppression of iNOS. These results may support the use of I. obliquus for food and medicinal application.

Sepsis, ensuing from unrestrained inflammatory replies to bacterial infections, endures with high injury and mortality worldwide. Presently, active sepsis management is missing in the hospitals during the surgery, and maintenance remnants mainly helpful. Now, we have constructed the macrophage bio-mimic nanoparticles for the treatment of sepsis and its management. Biomimetic macrophage nanoparticles containing a recyclable polymeric nanoparticle covered with cellular membrane resulting from macrophages (represented PEG-Mac@NPs) have an antigenic external similar to the cells. The PEG-Mac@NPs, Isorhamnetin (Iso) on the free LPS encouraged endotoxin in BALB/c mice through evaluating the nitric acid, TNF-α, and IL-6. Further, the COX-2 and iNOS expression ratio was examined to recognize the connection of several trails to find the exact mode of action PEG-Mac@NPs and Iso. The outcome reveals that the PEG-Mac@NPs inhibited and LPS triggered the NO production though the macrophages peritoneal. Furthermore, the anti-inflammatory possessions were additionally categorized through the reduction of COX-2 and iNOS protein expressions. Engaging PEG-Mac@NPs as a biomimetic decontamination approach displays potential for refining sepsis patient consequences, possibly in the use of sepsis management.

Fifteen previously undescribed coumarin derivatives, murratins A−M, including two pairs of coumarin enantiomers with a cyclopropane unit, two benzocoumarins, a dimeric coumarin, and eight C-8-substituted coumarins, together with six known analogues were isolated from the extract of the leaves and twigs of Murraya exotica L., a medicinal plant named “Jiulixiang” in Chinese. Their structures were elucidated on the basis of comprehensive analysis of 1D and 2D NMR and HRMS spectroscopic data, and the absolute configurations were assigned via comparison of the specific rotations, the ECD exciton coupling method, comparison of experimental and calculated ECD data, and the ECD data of in situ formed transition metal complexes. All the isolates were evaluated for their inhibitory effects on lipopolysaccharide-induced nitric oxide production in RAW 264.7 cells, and five coumarin derivatives showed moderate inhibitory activities. The possible mechanism for NO inhibition of undescribed bioactive compounds was deduced to interact with iNOS protein via molecular docking. The above results shed some light on a better understanding of the traditional anti-inflammation effect of M. exotica and coumarins are disclosed to be its potential anti-inflammatory constituents.

Chemical investigation of the EtOAc-soluble fraction from the MeOH/DCM extract of a cultured octocoral Briareum violaceum afforded two new briarane diterpenoids, briaviolides O (1) and P (2). The structures of briaranes 1 and 2 were elucidated on the basis of spectroscopic analysis, and 1 was shown to inhibit the accumulation of pro-inflammatory iNOS protein in LPS-stimulated RAW264.7 macrophage cells.

Our continuous search for new nitric oxide (NO) inhibitory substances as anti-neuroinflammatory agents for AD resulted in the isolation of one new labdane diterpenoid and three new guaiane sesquiterpenoids, as well as ten known compounds from Blumea balsamifera. Their structures were elucidated by NMR spectroscopic data analysis and the time-dependent density functional theory (TDDFT) electronic circular dichroism (ECD) calculations. The anti-neuroinflammatory effects were examined by inhibiting NO release in LPS-induced murine microglial BV-2 cells. The possible mechanism of NO inhibition of some bioactive compounds was also investigated using molecular docking, which revealed the interactions of bioactive compounds with the iNOS protein.

A phytochemical investigation to obtain new NO inhibitors resulted in the isolation of five new spiro diterpenoids (1 −5) from the aerial parts of Scutellaria formosana. The structures were elucidated on the basis of extensive 1D and 2D NMR spectroscopic data analysis, and the absolute configurations of these compounds were established via comparison of experimental and calculated electronic circular dichroism (ECD) spectra. The nitric oxide (NO) inhibitory effects were evaluated and all of the compounds showed inhibitory effects on lipopolysaccharide-induced NO production in murine microglial BV-2 cells. The possible mechanism of NO inhibition of bioactive compounds was also investigated using molecular docking, which revealed the interactions of bioactive compounds with the iNOS protein.

A phytochemical study on Scutellaria formosana afforded five previously undescribed spiro-diterpenoids, scutellapenes A−E. The structures were elucidated on the basis of extensive 1D and 2D NMR spectroscopic data analysis, and the absolute configurations of these compounds were established by the time-dependent density functional theory (TDDFT) electronic circular dichroism (ECD) calculations. Scutellapenes B−E possess a spiro-diterpenoid skeleton. All of the compounds showed inhibitory effects on LPS-induced nitric oxide (NO) production in murine microglial BV-2 cells. The further molecular docking studies revealed that these bioactive compounds had strong interactions with the iNOS protein.

A phytochemical investigation to obtain new NO inhibitors led to the isolation of nine compounds including one new guaiane-type sesquiterpenoid (1) and two new cleistanthane diterpenoids (2 and 3) from the stems of Trigonostemon howii. Their structures were elucidated on the basis of extensive 1D and 2D NMR spectroscopic data analysis, and the absolute configurations of new compounds 1–3 were established via comparison of experimental and calculated electronic circular dichroism (ECD) spectra. Compounds 2 and 3 possess a rare 3,4-seco-cleistanthane diterpenoid skeleton. All of the compounds showed inhibitory effects on lipopolysaccharide-induced NO production in murine microglial BV-2 cells. The further molecular docking studies indicated the strong interactions between some bioactive compounds with the iNOS protein, which revealed the possible and potential mechanism of NO inhibition of bioactive compounds.

Morinda officinalis How was widely applied to alleviate symptom like impotence, menstrual disorders, osteoporosis, and rheumatoid arthritis. To expand resources usage, phytochemistry of the aerial parts was studied and the structures of compounds were elucidated based on NMR, HRESIMS, IR and UV. Moreover, the anti-inflammatory effect and possible mechanism were investigated by Griess kit, RT-qPCR, ELISA, western blot and molecular docking on LPS-induced inflammation in RAW 264.7 cells. Herein, we isolated and identified 16 iridoid derivatives, including seven new iridoids officinaloside A-G (1-7) and nine known iridoids. All the compounds were safe to RAW 264.7 cells. Luckily, compounds 5 and 6 showed inhibitory effect on production of NO, and decreased the expression of inflammatory cytokines at mRNA and protein levels in a dose-dependent way. The possible mechanism of their anti-inflammation may be the affinity interaction between 5 with COX-2 protein, and 6 with iNOS protein. Overall, compounds 5 and 6 exert promising effects in inhibiting inflammatory cytokines, indicating that they could be used as lead compounds for developing health products or clinical practice for inflammation, which provides a scientific basis for further sustainable development and usage of the aerial parts of Morinda officinalis How.

Tussilago farfara L. is a well-known herb plant and used widely in China, North Africa, and Europe. A phytochemical investigation to obtain new NO inhibitors resulted in the isolation of three new sesquiterpenes, one new phenolic derivative, and ten known compounds from the flower buds of T. farfara . Their structures were established on the basis of extensive analyses of nuclear magnetic resonance (NMR) spectroscopic data. All of the isolates exhibited inhibitory effects on LPS-induced NO production in BV-2 cells. The possible mechanism of NO inhibition was also investigated using molecular docking, which revealed the interactions of bioactive compounds with iNOS protein. The present study disclosed that flower buds of T. farfara have the potential to be developed into a functional food.

Three new (1-3) and one known (4) bioactive terpenoids were isolated from the seeds of Silybum marianum based on the investigation to get new NO inhibitors. Their structures were determined by extensive NMR (1D and 2D NMR) and MS spectroscopic data, and the absolute configurations were identified by experimental and calculated ECD spectra. The NO inhibitory activities in murine microglial BV-2 cells and interactions with iNOS protein by molecular docking were evaluated for all compounds. The results showed that these compounds had potent NO inhibitory effects.

Five new compounds hostines A-E (1-5), together with eighteen known compounds (6-23), were obtained from the flowers of Hosta plantaginea (Lam.) Aschers, a traditional folk herbal medicine widely used in Mongolian. The structures of the undescribed compounds were determined by using detailed spectroscopic (1D/2D NMR) and HR-ESI-MS data analysis. The biological evaluation revealed that hostine C (3), hostine D (4), hostine E (5), stellarine (16), and phenethyl-O-β-d-glucopuranoside (19) possessed the significant anti-inflammatory activities, these compounds significantly inhibited the NO release of RAW 264.7 cells induced by LPS. The possible mechanism of NO inhibition of new bioactive compounds was also investigated using molecular docking, which revealed the interactions of bioactive compounds with the iNOS protein.

Porcine placenta extract (PPE) is known to possess anti-inflammatory properties owing to its high concentration of bioactive substances. However, the need to eliminate blood-borne infectious agents while maintaining biological efficacy raises concerns about the optimal method for sterilizing PPE. Therefore, the objective of this study was to compare the effects of the standard pressurized heat (autoclaving) method of sterilization with γ-irradiation on the anti-inflammatory effects of PPE. The anti-inflammatory actions of these two preparations of PPE were evaluated by measuring their inhibitory effects on the production of NO, the expression of iNOS protein, and the expression of iNOS, COX2, TNF-α, IL-1β, and IL-6 mRNA in lipopolysaccharide-stimulated RAW 264.7 cells. Compared with autoclaved PPE, γ-irradiated PPE showed significantly greater inhibition of NO production and iNOS protein expression, and produced a greater reduction in the expression of iNOS, COX2, TNF-α, IL-1β, and IL-6 mRNA. These results provide evidence that the sterilization process is crucial in determining the biological activity of PPE, especially its anti-inflammatory activity. Collectively, our data suggest that γ-irradiated PPE acts at the transcriptional level to effectively and potently suppresses the production of NO and the expression of pro-inflammatory cytokines.

PURPOSE: An attempt was made to characterize the orbital shear stress by comparing the effects of orbital shear stress on vascular endothelial cells (ECs) with the results of animal experiments. MATERIALS AND METHODS: In the laboratory study, cultured ECs of well were distinguished by center and periphery then exposed to orbital shear stress using an orbital shaker. In the animal study, arteriovenous (AV) fistulas were made at the right femoral arteries of Sprague-Dawley rats to increase the effect of the laminar flow. The condition of the stenosis was given on the left femoral arteries. The protein expression of inducible nitric oxide synthase (iNOS) and Akt phosphorylation were observed and compared. RESULTS: Under orbital shear stress, ECs showed an increase in iNOS protein expression and phosphorylation of Akt but most of the protein expressions derived from the periphery. When compared to the animal study, the increased expression of iNOS protein and phosphorylation of Akt were observed in the sample of AV fistula conditions and the iNOS protein expression was decreased in the stenosis conditions. CONCLUSION: Orbital shear stress did not show the characteristics of a pure turbulent shear force. By comparing the observation with the morphological changes of vascular ECs and site-specific protein expression on the results of animal experiments, uniform directional lamina shear stress forces were expressed at the periphery.

Two new briarane-type diterpenoids, briarenolides K (1) and L (2), were isolated from an octocoral identified as Briareum sp. The structures of new briaranes 1 and 2 were elucidated by spectroscopic methods. In the in vitro anti-inflammatory effects test, briaranes 1 and 2 were found to significantly inhibit the accumulation of the pro-inflammatory iNOS protein of the lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells.

A phytochemical investigation to search for new nitric oxide (NO) inhibitors resulted in the isolation of seven previously undescribed daphnane diterpenoids, thyrsoidpenes A−G, from the leaves of Trigonostemon thyrsoideus. Their structures including absolute configurations were elucidated on the basis of extensive NMR spectroscopic data analysis and the time-dependent density functional theory (TDDFT) electronic circular dichroism (ECD) calculations. Thyrsoidpenes B−G feature rare polycyclic caged structures of daphnane diterpenoid orthoester. The NO inhibitory effects were examined and all of the compounds showed inhibitory activities toward LPS-induced NO production in murine microglial BV-2 cells. The possible mechanism of NO inhibition of some bioactive compounds was also investigated using molecular docking, which revealed the interactions of bioactive compounds with the iNOS protein.

Citrus peel is a useful research resource containing important physiological function materials. In this study, anti-inflammatory effects of Citrus peel extracts were identified in LPS-induced macrophage cells. After the pretreatment of lipopolysarcharide (LPS), inhibitory effects of nitric oxide (NO), inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2) and pro-inflammatory cytokines, tumor necrotic factor-α (TNF-α), interleukin-6 (IL6), and interleukin-1 (IL-1), production were measured according to the treatment of citrus peel extracts. Study results showed approximately 40% inhibition of NO production but no concentration-dependent effects when citrus peel extracts were treated. According to the results of inhibitory effects of pro-inflammatory cytokines production, IL-6 had about 44% inhibition of production, TNF-α had about 13% inhibition of production, and IL-1 had no inhibition of production. According to western blot results, iNOS protein’s expression was slightly decreased. Also, COX-2 protein’s expression was observed to be slightly decreased in cells treated with citrus peels as iNOS’s result.

A phytochemical investigation to obtain new NO inhibitors resulted in the identification of six new (1–6) and four known (7–10) terpenoids from Salvia plebeia. Compounds 1 and 2 are new diterpenoids, 3–5 are new meroditerpenoids, 6–9 are sesquiterpenoids, and 10 is a known meroditerpenoid. The structures of these isolates were determined by routine NMR experiments and X-ray diffraction, as well as the electronic circular dichroism spectra. Compounds 1–4 are diterpenoids carrying an oxygen bridge, and 6 is a rare copane-type sesquiterpenoid with a bridged tricyclic framework. The isolates inhibited NO generation induced by lipopolysaccharide in BV-2 cells. The possible mechanism of NO inhibition of some bioactive compounds was also investigated using molecular docking, which revealed interactions of bioactive compounds with the iNOS protein.

Two new steroids, dendronesterones D (1) and E (2), featuring with 1,4-dienone moiety, along with three known steroids, methyl 3-oxochola-4,22-diene-24-oate (3), 5&alpha, 8&alpha, epidioxy-24(S)- methylcholesta-6,22-dien-3&beta, ol (4), and 5&alpha, epidioxy-24(S)-methylcholesta-6,9(11),22-trien-3&beta, ol (5), were isolated from an octocoral Dendronephthya sp. The structures of steroids 1 and 2 were elucidated by using spectroscopic methods and steroid 1 was found to exhibit significant in vitro anti-inflammatory activity in lipopolysaccharides (LPS)-induced RAW264.7 macrophage cells by inhibiting the expression of the iNOS protein.

Objectives : Stevia rebaudiana is a well-known herbal sweetener in the Korea, Japan and China, and its medical uses were originated from countries in South America. Although it has been shown the various medical effects of S. rebaudiana including contraception and treatment of human diseases such as hyperglycemia, it has almost not been studied about the efficacy of S. rebaudiana methanolic extract (SRE) on the acute inflammation and its action mechanism. Methods : To investigate the anti-inflammatory effects of SRE, we treated SRE and examined the level of inflammatory mediators in LPS-stimulated Raw264.7 cells. Results : Treatment of macrophage with LPS markedly induced the production of NO, and pro-inflammatory cytokines. Pretreatment of SRE blocked the induction of inflammatory mediators and the expression of iNOS protein. More importantly, LPS-induced phosphorylation of was suppressed by the treatment of SRE, suggesting SRE inhibition of NF- activation. Furthermore, SRE blocked LPS-induced phosphorylation of MAPKs. Conclusions : SRE inhibited the induction of NO, PGE2 and pro-inflammatory cytokines in Raw264.7 cells. SRE`s effect may be mediated with its inhibition of NF- activation and MAPK phosphorylation, which suggests its uses as an anti-inflammatory agents.

Five new eunicellin-based diterpenoids, krempfielins E–I (1–5) and seven known compounds (6–12) were isolated from the organic extract of a Taiwanese soft coral Cladiella krempfi. The structures of the new metabolites were elucidated on the basis of extensive spectroscopic analysis. Metabolites 5, 6, 10 and 12 were shown to exhibit cytotoxicity against a limited panel of cancer cell lines. Furthermore, compounds 6 and 10 could potently inhibit the accumulation of the pro-inflammatory iNOS protein, and 6 and 12 could significantly reduce the expression of COX-2 protein in LPS-stimulated RAW264.7 macrophage cells.

Three new steroidal carboxylic acids, paraminabic acids A–C (1–3) were isolated from a Formosan soft coral Paraminabea acronocephala. The structures of these compounds were established by extensive spectroscopic analysis and chemical methods. Application of the PGME method allowed the establishment of the absolute configurations at C-25 and C-24 for 1 and 2, respectively. Compound 3 showed potent cytotoxicity toward Hep3B, MDA-MB-231, MCF-7, and A-549 cancer cell lines, with IC50 values ranging from 2.05 to 2.83 μg/mL. Compounds 2 and 3 were found to inhibit the accumulation of the pro-inflammatory iNOS protein.

A phytochemical investigation to obtain new NO inhibitors resulted in the isolation of a new diterpenoid with a rare 9,10-seco-abietane skeleton (1) and twelve known terpenoids (2–13) from Callicarpa kwangtungensis. Their structures were elucidated on the basis of extensive 1D and 2D NMR spectroscopic data analyses, and the absolute configuration of compound 1 was established by comparison of the calculated and experimental electronic circular dichroism (ECD) spectra. The inhibitory activities on lipopolysaccharide-induced NO production in murine microglial BV-2 cells of these terpenoids were evaluated, and all of the compounds showed inhibitory effects. The following molecular docking studies showed interactions of the bioactive compounds with the iNOS protein.

A phytochemical investigation to obtain new NO inhibitors led to the isolation of two new ( 1 and 2 ) and four known ( 3 – 6 ) diterpenoids from Trigonostemon chinensis . Their structures were elucidated on the basis of extensive 1D and 2D NMR spectroscopic data analyses, and the absolute configurations of new compounds were established by experimental and calculated ECD spectra. The inhibitory activities on lipopolysaccharide-induced NO production in murine microglial BV-2 cells of these diterpenoids were evaluated, and all of the compounds showed inhibitory effects. The interactions of bioactive compounds with iNOS protein were also studied by molecular docking.

Salvia plebeiaR. Br. (Labiatae), distributed in many countries such as Korea, China, India, Iran, and Australia, is used as a folk remedy for a variety of inflammatory diseases including hepatitis, cough, diarrhea, gonorrhea, menorrhagia, tumors, and hemorrhoids. This study focused on determining the involvement of anti-inflammatory heme oxygenase-1 (HO-1) in the inhibitory activity of an extract of Salvia plebeiaR. Br. leaves (SPL) on nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in lipopolysaccharide (LPS)- stimulated RAW264.7 macrophages. SPL extract at the highest concentration (500 μg/mL) significantly inhibited NO production by approximately 85% and suppressed iNOS protein expression by approximately 90% compared to LPS-stimulated cells. The SPL extract induced the expression of HO-1 in a dose-dependent manner, and blocking HO-1 activity abolished the inhibitory effects of the SPL extract on NO production. These results suggest that an SPL extract has potent anti-inflammatory activity through HO-1 induction in RAW264.7 macrophages.

Series of carbamate and thiocarbamate derivatives were designed and synthesized and their inhibitory activities of NO production in lipopolysaccharide-activated macrophages were evaluated. Several thoicarbamate derivatives revealed promising inhibitory activity. The structure–activity relationship study of these compounds is also reported. Among these compounds, compound 12b was the most potent with 6.5 μM of IC50. They inhibited NO production through the suppression of iNOS protein and mRNA expression and nuclear translocation of p65.

Chrysanthemum zawadskii var. latilobum (CZ) has been widely used as a tea in Korea. In this study, we investigated the involvement of anti-inflammatory haem oxygenase-1 (HO-1) in the inhibitory activity of a CZ leaf extracts on nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. At the highest concentration of CZ leaf extract (50 μg/mL), NO production was 10% of that in LPS-stimulated macrophages, whereas iNOS protein expression was 5% of that in LPS-stimulated macrophages. CZ leaf extract also inhibited iNOS mRNA expression in a concentration-dependant manner, and at 50 μg/mL, iNOS mRNA expression was approximately 15% of that in LPS-stimulated cells. The CZ leaf extract induced HO-1 expression in a dose-dependent manner, and blocking HO-1 activity abolished the anti-inflammatory effect of CZ leaf extract. These results suggest that CZ leaf extract has a potent anti-inflammatory activity in RAW264.7 cells through the induction of HO-1.

Three new cembranoids, sarcocrassocolides M–O (1–3), have been isolated from the soft coral Sarcophyton crassocaule. The structures of the metabolites were determined by extensive spectroscopic analysis. Compounds 1–3 were shown to exhibit moderate cytotoxicity toward a limited panel of cancer cell lines and display significant in vitro anti-inflammatory activity in LPS-stimulated RAW264.7 macrophage cells by inhibiting the expression of the iNOS protein.

Seven new diterpenoids, namely, flexibilisolides C–G ( 1 – 5 ), flexibilisin C ( 6 ), and a novel 11,12-secoflexibillin ( 7 ), along with seven known compounds, 8 – 14 , were isolated from the Dongsha Atoll soft coral Sinularia flexibilis . The structures of the new metabolites were elucidated by extensive spectroscopic analysis and comparison of the NMR data with those of known analogues. Compounds 1 , 8 , and 11 were shown to exhibit moderate cytotoxic activity against HeLa and B16 cancer cell lines, and compound 10 was found to exhibit more potent cytotoxic activity against SK-Hep1 and B16 cancer cell lines. Moreover, compounds 1 , 2 , 8 , 9 , and 11 – 14 could significantly inhibit the accumulation of the pro-inflammatory iNOS protein and 1 , 8 , 11 , and 14 could reduce the accumulation of COX-2 protein in LPS-stimulated RAW264.7 macrophage cells.

Chemical investigations of the soft coral Sinularia gyrosa resulted in the isolation of six new norcembranolides, gyrosanolides A-F (1-6), a new norcembrane, gyrosanin A (7), and 11 known norditerpenoids 8-18. The structures of the isolated compounds were elucidated through extensive spectroscopic data and by comparison with reported data in the literature. Compounds 1-3, 7-9, 12, and 13 at concentration of 10microM did not inhibit the COX-2 protein expression, but significantly reduced the levels of the iNOS protein (55.2+/-14.6%, 18.6+/-6.7%, 10.6+/-4.6%, 66.9+/-5.2%, 10.2+/-5.1%, 17.4+/-7.2%, 47.2+/-11.9%, and 56.3+/-5.1%, respectively) by LPS stimulation. Compound 8 showed significant antiviral activity against HCMV (human cytomegalovirus) cells with an IC(50) of 1.9microg/mL.

Although it is documented that concurrent wounding increases mortality from radiation injury, the molecular mechanism of combined injury is unknown. In this study, mice were exposed to gamma radiation followed by skin wounding. Wound trauma exacerbated radiation-induced mortality, reducing the LD(50/30) from 9.65 Gy to 8.95 Gy. Analyses of histopathology, inducible nitric oxide synthase (iNOS), and serum cytokines were performed on mouse ileum and skin at various times after 9.75 Gy and/or wounding. In the ileum, the villi were significantly shortened 3 days postirradiation but not after wounding; combined injury resulted in decreased villus width and tunica muscularis thickness. The skin of mice subjected to combined injury was less cellular and had a smaller healing bud than the skin of mice subjected to wounding alone. Combined injury significantly delayed wound closure times; it also prolonged the increased levels of iNOS protein in the skin and ileum. iNOS up-regulation was correlated with increases in transcription factors, including NF-kappaB and NF-IL6. The increase in NF-IL6 may be due to increases in cytokines, including IL-1beta, -6, -8, -9, -10 and -13, G-CSF, eotaxin, INF-gamma, MCP-1, MIP-1alpha and MIP-1beta. Combined injury resulted in early detection of bacteria in the blood of the heart and liver, whereas radiation alone resulted in later detection of bacteria; only a transient bacteremia occurred after wounding alone. Results suggest that enhancement of iNOS, cytokines and bacterial infection triggered by combined injury may contribute to mortality. Agents that inhibit these responses may prove to be therapeutic for combined injury and may reduce related mortality.

Chemical investigations on the acetone extract of the Formosan soft coral Nephthea erecta have afforded a new calamenene-type sesquiterpene with a mercaptan group at C-15, erectathiol (1), and a previously reported sesquiterpenoid, (+)-trans-calamenene (2). A novel sec-germacrane sesquiterpene (3), along with a novel norergosterol, chabrosterol (4), possessing a 19-norergostane skeleton, was isolated from the other soft coral Nephthea chabroli. The structures of these metabolites were elucidated through extensive spectroscopic analyses. In vitro anti-inflammatory activity of 1 and 4 (10 μM) significantly reduced the levels of the iNOS protein (58.0 ± 6.5% and 12.4 ± 2.9%) and COX-2 protein (108.7 ± 4.5% and 45.2 ± 5.4%). In addition, metabolite 1 (166 μg/disk) exhibited antimicrobial activities against a small panel of bacterial strains.

Excessive production of nitric oxide (NO) generated by inducible nitric oxide synthase (iNOS) has been implicated in the pathogenesis of numerous inflammatory processes. The exact pathogenesis of denture-induced fibrous inflammatory hyperplasia (DIFIH) remains unclear. The aim of this study was to assess iNOS expression in healthy Oral mucosa (HOM) and DIFIH by immunohistochemical (IHC) method. The expression of iNOS protein in HOM has been reported in numerous publications; particularly these tissues were used as a control group. However, there are some conflicting results whether healthy Oral tissues express iNOS. In this study performing IHC method, DIFIH (n= 26) and HOM (n= 28) in formalin-fixed, paraffin-embedded sections were evaluated in terms of iNOS expression. The statistical analysis showed that there were no significant differences between DIFIH and HOM regarding iNOS immunoreactivity (p>0.05). The results in this study indicate that iNOS does not have a determining role in pathogenesi...

Five new steroids, griffinisterones A–E (1–5), were isolated from the octocoral Dendronephthya griffini. The structures of these compounds were elucidated by extensive spectroscopic analysis. The single-crystal X-ray crystallography on 1 allowed the determination of the 24R configuration in 1. The absolute stereochemistry of 5 was determined by the application of PGME method. Compounds 3 and 4 were found to significantly inhibit the accumulation of the pro-inflammatory iNOS protein of the LPS-stimulated RAW264.7 macrophage cells.

One new coumarin, 3,5-dihydroxy-7-O-α-L-rhamno pyranosyl-2H-chromen-2-one (1), was isolated from the whole plant of Sarcandra glabra. The structure was elucidated by spectroscopic methods. Our results indicated that 1 significantly inhibit nitric oxide (NO) production in LPS-induced RAW264.7 macrophages. RT-PCR analysis indicated it inhibited iNOS mRNA expression. In addition, Western blot analysis showed that 1 attenuated LPS-induced synthesis of iNOS protein in the macrophages. These results suggest that 1 could be potential anti-inflammatory agent by down-regulating iNOS expression.

S100beta is an astroglial-derived Ca2+ -binding protein having neurotrophic role on neurons and glial cells. An aberrant S100beta production has been observed in neurodegenerative disease, as Alzheimer's disease and Down syndrome. S100beta is responsible to start up a gliotic reaction by the release of pro-inflammatory mediators, including nitric oxide (NO) and cytokines from microglia and astrocytes, which are, in turn, deleterious for neurons. Interestingly, pro-inflammatory effect of S100beta seems not be restricted into the brain. Macrophages play a pivotal role in inflammatory diseases, occurring both in the brain and in the periphery. In this study, we tested the hypothesis that S100beta may affect macrophage functions, amplifying thus the inflammatory process. Our results demonstrate that S100beta stimulates both NO production and iNOS protein transcription and expression in J774 and rat peritoneal macrophages. NO production was concentration and time-dependently inhibited by two iNOS inhibitors, L-NAME and SMT. We also demonstrated that S100beta induced oxidative stress by increasing H2O2 production and lipid peroxidation of cell membrane in both macrophage types. The pro-oxidant potential of S100beta activates p38 MAP kinase (MAPK), which has been described to directly activate NF-kappaB. In our study, SB203580, a p38 MAPK inhibitor, and two NF-kappaB inhibitors, TLCK and BAY 11-7082, decreased both NO production and iNOS protein transcription and expression in S100beta-stimulated J774 and peritoneal rat macrophages. Moreover, additional studies demonstrated that S100beta affected also TNF-alpha protein expression in J774 macrophages. In conclusion, our results highlight the potential role of S100beta during an inflammatory scenario identifying macrophages as a novel S100beta-responsive cell-type.

The effect of fermentation on the nitric oxide (NO)-suppressing activity of tea was investigated using an lipopolysaccharide (LPS)-activated RAW 264.7 cell model. Two species of tea, Taiwan tea No. 12 and Chinsin Oolong, commonly cultivated and consumed in Taiwan, were selected and manufactured with three degrees of fermentation. All of the teas including non-fermented green, partially fermented paochong and fully fermented black teas, were prepared from a single batch of fresh tea leaves. Additionally, the freeze-dried solids of tea infusions were used to treat cells and their NO-suppressing activities were compared. The results showed that the two species of black tea inhibited the NO production, inducible nitric oxide synthase (iNOS) catalytic activity and iNOS protein expression in LPS-activated cells, more weakly than did green and paochong teas. Meanwhile, the NO-suppressing activity was highly correlated with the total phenolics content. Clearly, heavy fermentation strongly affects the NO-suppressing activity of tea, and the decline of the NO-suppressing activity is attributed to the elimination of the phenolics.

We investigated the effects of two NOS inhibitors (AG and l -NAME) on DMBA-induced hamster buccal–pouch carcinogenesis. Six hundred Syrian golden hamsters were split into two divisions (I and II); divisions split into three groups (experimental groups A and B, control group C); and each group into subgroups of 20 (A1–A6, B1–B6 and C1–C3). The pouches of animals in groups A1–A3 were painted first with AG of differing concentrations (10, 20, and 30 μmol/ml) and then 30 min later with DMBA (0.5%), thrice weekly for 9 weeks. Subgroups A4–A6 only received AG treatment. Groups B1 to B6 were similarly treated with l -NAME. Animals in division II were treated in the same manner for 13 weeks. Post-mortem analysis revealed that both inhibitors can suppress the development of epithelial dysplasias and squamous-cell carcinomas. An associated increase in the numbers of epithelial hyperplasias was paralleled by a decrease in iNOS protein expression. This animal model can be employed to evaluate the potential use of iNOS inhibitors as novel therapeutic tools for oral squamous-cell carcinogenesis.

In this report, our findings highlighted the presence of a high level of calcium in the cortex following traumatic brain injury (TBI) in a rat model of fluid percussion-induced brain injury. This calcium increase represents a pitfall in the assessment of Ca 2+ -independent nitric oxide synthase (NOS) activity supposed to play a role in the secondary brain lesion following TBI. The so-called Ca 2+ -independent NOS activity measured in the injured cortex 72 h after TBI had the pharmacological profile of a Ca 2+ -dependent NOS and was therefore inhibited with a supplement of calcium chelator. The remaining activity was very low and iNOS protein was hardly immunodetected on the same sample used for NOS activity assay. The concentration of calcium chelator used in the assay should be revised and adjusted consequently to make sure that the calcium-free condition is achieved for the assay. Otherwise, the findings tend towards an overestimation of Ca 2+ -independent and underestimation of Ca 2+ -dependent NOS activities. The revised Ca 2+ -independent NOS activity assay was then tested, in relation with the amount of iNOS protein, in a model of LPS-induced neuroinflammation. Taken together, precautions should be taken when assessing the Ca 2+ -independent enzymatic activity in cerebral tissue after a brain insult.