Induced Mouse Ear Research Articles

Anti-inflammatory activity of Hyptis albida

Anti-inflammatory activity of chloroform and methanol extracts from aerial parts of Hyptis albida was tested on different pharmacological models in rats and mice. The plant was collected in Guadalcazar San Luis Potosi State, dried and powdered. The extracts were obtained by heating the solvent to its boiling point. The extracts free of solvent were tested in two pharmacological models, carrageenan-induced edema and 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced mouse ear edema. In the case of carrageenan model, the extract was administered orally using polyvinylpyrrolidone (PVP) like vehicle and for the TPA, topical administration was used. Results showed that chloroform and methanol extracts had effects on TPA-induced edema, the major effect was observed with the chloroform extract. Also this extract inhibited edema cause by multiple applications of TPA (53.2%). In carrageenan induced edema test, the chloroform extract showed no dose dependent response at doses of 100, 200, and 400 mg/kg. The results suggest that H. albida could be a source for compounds with anti-inflammatory effect. Key words: Hyptis albida, anti-inflammatory activity, 12-O-tetradecanoylphorbol 13-acetate (TPA), carragenan

Antioxidant and anti-inflammatory activities of Pittocaulon species from México

Context: Stems and leaves of Pittocaulon spp. (Asteraceae) are used in Mexican traditional medicine as an anti-inflammatory substance and for the treatment of skin injuries.Objective: This study evaluated the antioxidant activity of methanol (MeOH) and dichloromethane (DC) extracts of five Pittocaulon species.Materials and methods: DC and MeOH extracts from flowers, roots, and stems of Pittocaulon praecox (Cav.) H. Rob. & Brettell, P. bombycophole (Bullock) H. Rob. & Brettell, P. filare (Mc Vaugh) H. Rob. & Brettell, P. velatum (Greenm.) Rob. & Brettell and P. hintonii H. Rob. & Brettell.Result and Discussion: In the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, the flower extracts obtained with MeOH were the most active with IC50 values ranging from 51.83 ± 4.08 to 154.19 ± 8.39 ppm. In the thiobarbituric acid reactive substances (TBARS) model, the best activity was shown by DC extracts of roots with IC50 values ranging from 55.54 ± 1.28 to 160.82 ± 5.37 ppm. The MeOH extract of flowers of P. bombycophole had the highest IC50 value in both DPPH (51.83 ± 4.08 ppm) and TBARS (39.78 ± 1.97 ppm). The samples with the best values in the antioxidant activity assays were evaluated in the anti-inflammatory tests. The DC root extract of P. velatum at a dose of 1 mg/ear produced the greatest reduction (84.96%) of the 2-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse ear edema. This extract also reduced the activity of the enzyme myeloperoxidase (MPO) (73.65%) at the same dose. In contrast, DC root extract of this species did not show significant inhibition of the increase in paw edema induced by carrageenan at the doses tested (100 mg/kg).Conclusion: These results support the traditional use of these plants as anti-inflammatory. DC extracts of P. velatum and MeOH extracts of P. bombycophole may be a potential resource of natural anti-inflammatory and antioxidant compounds, respectively. Additional studies must be done to identify the compounds responsible of the activity on these plants and to establish the mechanism of action.

The different inhibitory effects of Huang-Lian-Jie-Du-Tang on cyclooxygenase 2 and 5-lipoxygenase

BackgroundHuang-Lian-Jie-Du-Tang (HLJDT), a famous traditional Chinese prescription with wide anti-inflammatory applications, is an aqueous extract of four herbal materials: Rhizoma coptidis, Radix scutellariae, Cortex phellodendri, and Fructus gardeniae. Its effects on the cyclooxygenase (COX)-2 and 5-lipoxygenase (5-LOX) pathways are thought to be responsible for its anti-inflammatory activity. However, our previous work found that the inhibitory effects of HLJDT act on the 5-LOX pathway but not on the COX pathway. The possibility that HLJDT inhibits COX-2- or 5-LOX-catalyzed eicosanoid generation by downregulating enzyme expression requires further investigation. Aim of the studyTo observe the effects of HLJDT and its four major components (baicalin, baicalein, berberine and geniposide) on COX-2- or 5-LOX-catalyzed eicosanoid generation and to distinguish the effects of HLJDT on enzyme activity from those on enzyme expression. MethodsThe topical anti-inflammatory activities and inhibition of eicosanoid formation of HLJDT and its components were observed in an arachidonic acid (AA)-induced mouse ear edema model. Macrophage-based systems were established to observe the effects of the drugs on enzyme activity and enzyme expression of COX-2 and 5-LOX. Further experiments were carried out to confirm these effects at the mRNA and protein levels. ResultsTopical treatment of HLJDT significantly inhibited AA-induced mouse ear edema and reduced PGE2 and LTB4 release in the edematous ears. Baicalein, geniposide, and berberine also ameliorated the symptoms and suppressed eicosanoid generation with varying efficacies. Cell-based assays showed that HLJDT and baicalein inhibited the PGE2 levels by decreasing COX-2 enzyme expression without affecting COX-2 enzyme activity in RAW 246.7 murine macrophages. The other experiments on rat peritoneal macrophages indicated that HLJDT and baicalein exerted significant inhibition on LTB4 production by decreasing 5-LOX enzyme activity. The real-time PCR and western blotting data demonstrated that HLJDT and baicalein reduced COX-2 expression at the mRNA and protein levels, whereas no inhibition on 5-LOX expression was observed. ConclusionsHLJDT can suppress eicosanoid generation via both the COX and LOX pathways, which definitely contributes to its topical anti-inflammatory activity. We have confirmed that its dual inhibition on the COX and LOX pathways mainly result from the downregulation of COX-2 expression and direct inhibition of 5-LOX activity, respectively. Baicalein worked as a potent active component in most of the tests. These findings about the different inhibitory effects of HLJDT on COX-2 and 5-LOX help to better understand the mechanism of HLJDT and promote safer applications of drug.

Kramecyne--a new anti-inflammatory compound isolated from Krameria cytisoides.

In the present work we describe the structure and anti-inflammatory activity of a new compound, kramecyne, isolated from a methanol extract of Krameria cytisoides (Krameriaceae). The structure of kramecyne was determined by IR, NMR, MS, and elemental analysis, which indicated that the structure corresponded to a hexamer of cyclic peroxide monomers. This compound exhibited good anti-inflammatory activity in the 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse ear edema (51.8 ± 6.9% inhibition) and carrageenan-induced rat paw edema models at doses of 50 mg/kg. The compound significantly reduced edema to 63.1% after 1.0 h, and the effect was unchanged for 5 h. Kramecyne did not present acute toxicity, even at doses of 5,000 mg/kg.

Ethnopharmacological evaluation of some Scorzonera species: In vivo anti-inflammatory and antinociceptive effects

Ethnopharmacological relevanceScorzonera species are mainly used against inflammation and to relieve pain in Turkish traditional medicine. Therefore, we aimed to assess in vivo anti-inflammatory and antinociceptive activities of the aerial part and root extracts of Scorzonera acuminata, Scorzonera cana var. alpina, Scorzonera cana var. jacquiniana, Scorzonera cana var. radicosa, Scorzonera cinerea, Scorzonera eriophora, Scorzonera incisa, Scorzonera laciniata ssp. laciniata, Scorzonera parviflora and Scorzonera sublanata. Materials and methodsFor the anti-inflammatory activity evaluation carrageenan, PGE2 and serotonin-induced hind paw edema and 12-O-tetradecanoyl-13-acetate (TPA)-induced mouse ear edema models were used. p-Benzoquinone-induced abdominal constriction test was employed in mice for the assessment of antinociceptive activity. Furthermore chemical composition of the tested extracts was investigated qualitatively and quantitatively by using RP-HPLC method. Some phenolic acids and flavonoids were used as standards. ResultsExtracts prepared from the aerial parts of Scorzonera cana var. jacquiniana, Scorzonera cinerea, Scorzonera eriophora, Scorzonera incisa and Scorzonera parviflora showed significant inhibitory effect on carrageenan and PGE2-induced hind paw edema model as well as on p-benzoquinone-induced abdominal constriction test. The extracts did not show any remarkable activity on serotonin-induced hind paw edema and TPA-induced mouse ear edema models. Chlorogenic acid was detected as major compounds in all the species investigated. Additionally, among the tested flavonoids, luteolin-7-glucoside, hyperoside and rutin were found to be in different amounts in Scorzonera species. ConclusionThe experimental data revealed that Scorzonera cana var. jacquiniana, Scorzonera cinerea, Scorzonera eriophora, Scorzonera incisa and Scorzonera parviflora possess significant anti-inflammatory and antinociceptive activity. It has been suggested that flavonoids and chlorogenic acid are partly responsible for mentioned activities of Scorzonera species.

Anti-inflammatory activities of fractions from Geranium nepalense and related polyphenols

Geranium nepalense Sweet is a common Chinese herbal medicine and has been used as influenza, dysentery, antiphlogistic and analgesic tonic, hemostatic, stomachic, and antidiabetic drugs. The anti-inflammatory effects of G. nepalense on tetradecanoyl phorbol acetate (TPA)-induced mouse ear edema were studied in this work. The results showed that ethyl acetate fraction of the water extract of G. nepalense possessed significant activity at 2.5 g/kg (p < 0.01) with aspirin as a positive control (0.6 g/kg). Six polyphenolic compounds, including three flavonoids, i.e. kaempferol, kaempferol-7-O-β-D-glucopyranoside, and quercetin-7-O-α-rhamnopyranoside, and two tannins, i.e. pyrogallol and gallic acid, and one lignin, i.e. epipinoresinol, were isolated and characterized from ethyl acetate fraction. The isolation of polyphenols provides a clue for beneficial effects of G. nepalense in the demonstrated anti-inflammatory activity.

Effects of phycocyanin extract on prostaglandin E2 levels in mouse ear inflammation test.

Recently it was demonstrated that phycocyanin, a biliprotein isolated from microalgae Spirulina, exerts anti-inflammatory activity in several animal models of inflammation. In this report, the effects of phycocyamin on prostaglandin E2 (PGE2) concentrations and phospholipase A2 (PLA2) activity were determined in arachidonic acid (AA) and 12-O-tetradecanoyl phorbol 13-acetate (TPA)-induced mouse ear oedema, respectively. Phycocyanin (50-200 mg/kg p.o.) inhibited in a dose-dependent manner PGE2 levels in mouse ear treated with AA. Also, phycocyanin (100-400 mg/kg p.o.) moderately reduced PLA2 activity in TPA-induced mouse ear inflammation test. In this model triamcinolone (10 mg/kg p.o.) used as reference drug exerted a remarkable inhibitory effect on PLA2 activity. These results provide the first evidence that the anti-inflammatory effects of phycocyanin may result, at least partially, from inhibition of PGE2 production and a moderate inhibition of PLA2 activity.

Attenuation of experimental TPA-induced dermatitis by acetylenic acetogenins is associated with inhibition of PLA 2 activity

Topical application of acetylenic acetogenins (AAG) from avocado (0.01–1.0 mg/ear), was effective in inhibiting both 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse ear edema, and in decreasing tissue myeloperoxidase activity (indicative of polymorphonuclear leukocyte influx). Maximum edema inhibition of 72% was achieved by AAG at lower concentration (0.6 mg/ear) than that of the anti-inflammatory drug indomethacin (2 mg/ear). The maximum myeloperoxidase inhibition of 60% was obtained at AAG concentration 0.1 mg/ear. Chemical reduction of unsaturated bonds in aliphatic chain of AAG molecules almost abrogated inhibition effect of AAG at high concentration. In vitro AAG administration reduced secretion of PGE 2 in TPA-induced keratinocytes, and inhibited total PLA 2 and sPLA 2 activities in HaCaT cells. The results indicate a topical anti-inflammatory effect of acetylenic acetogenins which is associated with inhibition of PLA 2 activity in skin.

Effectiveness of Vernonia scorpioides ethanolic extract against skin inflammatory processes

Ethnopharmacological relevanceVernonia scorpioides (Asteraceae) is a native Brazilian medicinal plant that is commonly used to treat skin disorders. Considering the traditional use of Vernonia scorpioides and the lack of information about its pharmacological properties, we investigated the topical anti-inflammatory effect of the ethanolic extract of Vernonia scorpioides (EEVS) on acute and chronic cutaneous inflammation models in mouse. Materials and methodsThe topical anti-inflammatory effect of EEVS was evaluated against acute models (12-O-tetradecanoylphorbol acetate (TPA)- and arachidonic acid (AA)-induced mouse ear oedema) and chronic models (multiple applications of croton oil). ResultsThe EEVS caused a dose-related inhibition of oedema in both the TPA- and AA-induced acute models (DI50=0.24 and 0.68mg/ear with an inhibition of 80±5% and 65±5%, respectively, for 1mg/ear). In addition, the TPA-induced increase in myeloperoxidase activity (MPO) in the ear was reduced (77±8%) by the topical application of EEVS. In the chronic model, the EEVS reduced all parameters evaluated: oedema formation (31±2%), epidermal hyperproliferation (histology) and MPO (25±10%). However, the topical treatment of EEVS had no effect on N-acetyl-β-d-glucosaminidase activity. The EEVS effectively interfered in the ear oedema on the delayed-type hypersensitivity reaction induced by oxazolone. The topical treatment with EEVS performed on both phases or only on the elicitation phase caused the inhibition of the ear oedema-induced by oxazolone in 42.9% and 63.4%, respectively, when compared to control animals (sensitized and challenged). ConclusionsThe results suggest that EEVS is effective as a topical anti-inflammatory agent in acute and chronic inflammatory processes and that its action is markedly influenced by the inhibition of neutrophil migration into inflamed tissue as well as by epidermal hyperproliferation.

Discovery of novel anti-inflammatory drug-like compounds by aligning in silico and in vivo screening: The nitroindazolinone chemotype

In this report, we propose the combination of computational methods and in vivo primary screening in zebrafish larvae and confirmatory in mice models as a novel strategy to accelerate anti-inflammatory drug discovery. Initially, a database of 1213 organic chemicals with great structural variability – 587 of them anti-inflammatory agents plus 626 compounds with other clinical uses – was divided into training and test groups. Atom-based quadratic indices – a TOMOCOMD- CARDD molecular descriptors family – and linear discriminant analysis (LDA) were used to develop a total of 13 models to describe the anti-inflammatory activity. The best model (Eq. (13)) shows an accuracy of 87.70% in the training set, and values of Matthews correlation coefficient ( C) of 0.75. The robustness of the models was demonstrated using an external test set as validation method, i.e., Eq. (13) revealing classification of 88.44% ( C = 0.77) in this series. All models were employed to develop ensemble a QSAR classification system, in which the individual QSAR outputs are the inputs of the aforementioned fusion approach. The fusion model was used for the identification of novel anti-inflammatory compounds using virtual screening of 145 molecules available in our in- house library of indazole, indole, cinnoline and quinoxaline derivatives. Out of these, 34 chemicals were selected, synthesized and tested in a lipopolysaccharide (LPS)-induced leukocyte migration assay in zebrafish larvae. This activity was evaluated based on leukocyte migration to the injury zone of tail-transected larvae. Compounds 18 (3 μM), 24 (10 μM), 25 (10 μM), 6 (10 μM), 15 (30 μM), 11 (30 μM) and 12 (30 μM) gave the best results displaying relative leukocyte migration (RLM) values of 0.24, 0.27, 0.35, 0.41, 0.17, 0. 26 and 0.27 respectively, date that suggest an anti-inflammatory activity of 76, 73, 65, 59, 83, 84 and 73%, respectively. Compound 18 was the most potent but showed high toxicity together with compound 6. Next, we used the tetradecanoylphorbol acetate (TPA)-induced mouse ear oedema model to evaluate the most potent compounds in the zebrafish larvae tail transection assay. All assayed compounds, with the exception of chemical 15, showed anti-inflammatory activity in mice. Compound 12 ( VA5- 13l, 2-benzyl-1-methyl-5-nitro-1,2-dihydro-3 H-indazol-3-one) was the most active and completely abolished the oedema. Compounds 6, 11 and 24 showed inhibition percentages in the range of the reference drug (indomethacin), whereas compounds 18 and 25 reduced the oedema in a lesser extent (inhibition of 73 and 80%, respectively). In addition, all compounds except chemical 15, significantly reduced neutrophil infiltration, measured as myeloperoxidase activity on TPA application test. Compounds 6, 11, 12 and 18 showed values comparable to indomethacin (inhibition percentage of 61), but compounds 6 and 18 were toxic in zebrafish and showed unspecific cytotoxicity in murine macrophages at 100 μg/mL, while the remaining compounds 11, 12 and 25 were inactive at most levels. Evidently, this study suggests a new support structure ( 12, 11 and 24; a nitroindazolinone chemotype) that constitutes a novel promising lead and may represent an important therapeutic alternative for the treatment of inflammatory conditions.

The aqueous root extract of Aristolochia ringens (Vahl.) prevents chemically induced inflammation

Aristolochia ringens Vahl Aristolochiaceae belongs to a family with many medicinal uses, but also reported toxic (1). Based on its use in traditional medicine, the antiinflammatory activity of the aqueous root extract of Aristolochia ringens (AR) (10–100mg/kg p.o.) was evaluated using the carrageenaan and egg albumin induced rat paw oedema (2), formaldehyde induced arthritic inflammation (3) and xylene induced mouse ear oedema methods (2). AR (10–50mg/kg) produced a dose-dependent decrease in rat paw oedema in the carrageenan and egg albumin induced inflammations at the time intervals studied. The maximum inhibitory effects of AR (50mg/kg), 57.1% and 65.6% in both experiments were comparable to the 57.9% and 63.9% of standard drugs, indomethacin and diclofenac (10mg/kg p.o.) respectively. AR (10–50mg/kg) also dose dependently inhibited the arthritic paw oedema induced by formaldehyde over the 10 day period of study. The maximum inhibition by AR (50mg/kg) (50%) was greater than the 40.8% inhibition by diclofenac (10mg/kg i.p.). AR (10–50mg/kg) also produced a significant (p<0.05) dose dependent inhibition of mouse ear oedema, with a peak effect at 50mg/kg of 84.78%, which was greater than the 65.21% inhibition by dexamethasone (1mg/kg). No mortality was observed in 24 hours, with AR (up to 10g/kg p.o.), but an LD50 of 453mg/kg was obtained with the intraperitoneal route of administration in mice. Results suggest that the aqueous root extract of Aristolochia ringens possesses antiinflammatory activity, inhibiting chemically induced inflammation.

Embelin reduces cutaneous TNF-α level and ameliorates skin edema in acute and chronic model of skin inflammation in mice

Tumor necrosis factor-α (TNF-α) is known to play a crucial role in the pathogenesis of psoriasis. The present study was designed to investigate the effects of embelin on lipopolysachharide induced TNF-α production in mice and in human keratinocytes in vitro and also to study the effect of embelin on acute and chronic skin inflammation in mice. Production of pro-inflammatory cytokines (TNF-α and IL-1β), activation of myeloperoxidase and histological assessment were examined in acute and chronic skin inflammation using 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced mouse ear edema. Embelin inhibited topical edema in the mouse ear, leading to substantial reductions in skin thickness and tissue weight, inflammatory cytokine production, neutrophil-mediated myeloperoxidase activity, and various histopathological indicators. Furthermore, embelin was effective at reducing inflammatory damage induced by chronic TPA exposure. Our data indicate that embelin has anti-inflammatory activities in both acute and chronic irritant contact dermatitis in vivo and this effect of embelin may be due, at least in part, to the inhibition of IL-1β and TNF-α and to the subsequent blockade of leukocyte accumulation.

Anti-inflammatory activity in rats and mice of phenolic acids isolated from Scrophularia frutescens.

Different species of the Scrophularia genus (Scrophulariaceae) have been reported to have bacteriostatic and anti-inflammatory properties. In previous studies the anti-inflammatory and antibacterial activity of different extracts from Scrophularia frutescens were investigated and p-coumaric, caffeic, ferulic gentisic, protocatechuic, syringic and isovanillic acids were isolated and identified. In this work the anti-inflammatory activity of these compounds, administered orally, has been studied against carrageenan-induced rat paw oedema and, administered topically, against tetradecanoylphorbol acetate (TPA)-induced mouse ear oedema. The compounds' myeloperoxidase activity in inflamed ear was also investigated. Some of the phenolic acids were remarkably active in the TPA test (protocatechuic 71.59% inhibition, P < 0.001; syringic 74.43%, P < 0.001; ferulic 71.02% P < 0.001) and all significantly inhibited mouse ear oedema. They were only moderately active, or were without activity, in the carrageenan test. These results imply that the phenolic acids assayed are more effective topically than as oral anti-inflammatory agents and that their action is markedly influenced by the inhibition of neutrophil migration into inflamed tissue. This study has also enabled us to make some observations on the possible relationship between the chemical structure and anti-inflammatory activity of the compounds assayed.

In Vivo Activity of Benzoyl Ester Clerodane Diterpenoid Derivatives from Dodonaea polyandra

Four new benzoyl ester clerodane diterpenoids, 15,16-epoxy-8α-(benzoyloxy)methylcleroda-3,13(16),14-trien-18-oic acid (1), 15,16-epoxy-8α-(benzoyloxy)methyl-2α-hydroxycleroda-3,13(16),14-trien-18-oic acid (2), 15,16-epoxy-8α-(benzoyloxy)methyl-2-oxocleroda-3,13(16),14-trien-18-oic acid (3), and 15,16-epoxy-2α-benzoyloxycleroda-3,13(16),14-trien-18-oic acid (4), have been isolated from the leaves and stems of Dodonaea polyandra. The anti-inflammatory activities of compounds 1, 2, and 4 were evaluated by means of 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse ear edema. Compounds 2 and 4 exhibited maximum inhibition of inflammation (70-76%) at doses of 0.22 and 0.9 μmol/ear, respectively. Modest activity (~45% inhibition) was maintained at nanomole/ear doses.

Anti-Inflammatory and Cytotoxic Activities of Chichipegenin, Peniocerol, and Macdougallin Isolated from Myrtillocactus geometrizans (Mart. ex Pfeiff.) Con.

The oleanane-type triterpene chichipegenin and the sterols peniocerol and macdougallin, isolated from Myrtillocactus geometrizans, showed anti-inflammatory activities in both the 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse ear edema model and the carrageenan-induced rat paw edema model. All tested compounds inhibited the TPA-induced edema in a dose-dependent manner, with ED50 values less than or equal to that shown by indomethacin. Among them, peniocerol was the most active compound. However, only peniocerol and macdougallin reduced carrageenan-induced rat paw edema. On the other hand, peniocerol and macdougallin showed cytotoxicity against several human cancer cell lines. These results indicate that compounds isolated from M. geometrizans possess antiinflammatory and cytotoxic properties, and the presence of chichipegenin in the aerial parts could justify the medicinal uses attributed to the plant.

Anti-inflammatory effect of Lithrea molleoides extracts and isolated active compounds

In this study the anti-inflammatory activity of aqueous, dichloromethane (CH₂Cl₂) and methanolic (MeOH) extracts and two major compounds isolated from Lithrea molleoides (Vell.) Engl. (Anacardiaceae) were evaluated. Two classical experimental models were used, carrageenan-induced rat paw edema and 12-O-tetradecanoylphorbol-13 acetate (TPA) induced mouse ear edema. MeOH extracts exhibited a significant systemical anti-inflammatory effect in the carrageenan (inhibition of 46% at 3 h) and in the TPA-ear edema test (inhibition of 21%). The presence of methyl gallate (inhibition of 63% in TPA ear edema), as one of the main compounds in the active fraction from MeOH extract may be explained the effect observed. Also, 1,3-dihydroxy-(Z,Z)-5-(tridec-4',7́dienyl) benzene obtained from CH₂Cl₂ extract showed a significant topical anti-inflammatory activity (inhibition of 68%). Furthermore, no signs of toxicity were observed with doses up to 3g/kg in an acute toxicity assay. The results of this study present evidence that Lithrea molleoides given either systemically or topically has anti-inflammatory properties.

Anti-inflammatory effects of Glehnia littoralis extract in acute and chronic cutaneous inflammation

Glehnia littoralis (Umbelliferae) is a traditional medicine used in Korea, China, and Japan to treat the immune-related diseases. However, its anti-inflammatory activities and mechanisms remain to be defined. We investigated the effects of 70% ethanolic extract from G. littoralis (GLE) on skin inflammation in mice. Production of proinflammatory cytokines (IL-1β and TNF-α), activation of myeloperoxidase (MPO), and histological indicators were examined in acute and chronic skin inflammation using 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced mouse ear edema. We also performed acetic acid-induced vascular permeability tests. GLE treatment at 200 mg/kg inhibited topical edema in the mouse ear, leading to substantial reductions in skin thickness and tissue weight, inflammatory cytokine production, neutrophil-mediated MPO activity, and several histopathological indicators. Furthermore, GLE effectively reduced inflammatory damage induced by chronic TPA exposure and significantly inhibited the vascular permeability induced by acetic acid in mice. These results suggest that G. littoralis is an effective anti-inflammatory agent in murine phorbol ester-induced dermatitis and may have therapeutic potential in a variety of immune-related cutaneous diseases.

Synthesis of N-Pyridinyl(methyl)-1,2-dihydro-4-hydroxy-2-oxoquinoline-3-carboxamides and analogues and their anti-inflammatory activity in mice and rats

The topical anti-inflammatory activity of a series of N-pyridinyl(methyl)1,2-dihydro-4-hydroxy-2-oxoquinoline-3-carboxamides, analogues of roquinimex, has been evaluated by measuring their inhibitory effect in the phorbol myristate acetate (PMA)-induced mouse ear swelling test, used as a screening test. All the eight carboxamides tested (9-16) exhibited significant inhibitory activity at 0.4 and 0.2 mM kg(-1). The most potent compound, the 6-bromo derivative 12, induced a 73% inhibition at 0.2 mM kg(-1). Pharmacomodulation was carried out by heterocycle opening and molecular simplification leading to pentafluorobenzoylacetamide 17, pentafluorocinnamamides 18 and 19, and pentafluorobenzaldimines 20 and 21. All the five compounds exerted a reduction in swelling (49-63% at 0.2 mM kg(-1)) comparable with ibuprofen (56%). Anti-inflammatory activity of the most efficient compounds was evaluated by carrageenan-induced rat paw oedema inhibition. The pentafluorobenzaldimine 20 showed the highest activity with an inhibition percentage of 85% at 0.2 mM kg(-1).

In-vitro and in-vivo immunomodulatory effects of syringin.

Syringin was found to possess immunomodulatory activity by which it inhibited the in-vitro immunohaemolysis of antibody-coated sheep erythrocytes by guinea-pig serum through suppression of C3-convertase of the classical complement. In this study, we examined its in-vitro and in-vivo activity on tumour necrosis factor (TNF)-alpha and nitric oxide (NO) production, CD4+ T cell and CD8+ cytotoxic T cell (CTLL-2) proliferation, and croton oil-, arachidonic acid- and fluorescein-isothiocynate (FITC)-induced mouse ear oedema model. Syringin significantly inhibited both TNF-alpha production from lipopolysaccharide (LPS)-stimulated RAW264.7 cells and CD8+ T cell (CTLL-2) proliferation in a dose-dependent manner, whereas neither NO production nor CD4+ T cell proliferation were blocked even by high concentrations of syringin. In the invivo experiments, syringin also significantly suppressed FITC-induced ear oedema in mice but not the ear oedema induced by croton or arachidonic acid. These results suggest that syringin may be implicated as an immunomodulator having an anti-allergic effect rather than an anti-inflammatory effect. The anti-allergic effect of syringin seems to be due, in part, to inhibition of TNF-alpha production and cytotoxic T cell proliferation.

Synergistic Effect of Notopterygium incisum with Clematis manshurica in the Anti-inflammatory Activity

Objective : Oriental medicines have been combined oriental medical theory which based on the seven modes of emotions. Notopterygium incisum (N. incisum) and Clematis manshurica (C. manshurica) have been used as an anti-rheumatic and analgesic medicine for the treatment of rheumatism, headache, cold, etc. In this study, we evaluate the synergistic anti-inflammatory effect of N. incisum and C. manshurica. Method : To evaluate the synergistic anti-inflammatory effect of a herbal mixture N. incisum and C. manshurica, we examined the changed ear thickness in 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced mouse ear edema model after topical application of herbal mixtu re. In addition, the levels of markers for inflammation, such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, prostaglandin G2 (PGE2 ), and nitric oxide (NO) were measured by ELISA assay and Griess reagent in lipopolysaccharide-stimulated Raw 264.7 cells. Results : Our results showed that aqueous extracts of N. incisum and C. manshurica combination significantly inhibited the mouse ear edema induced by TPA. Moreover, the aqueous extracts of N. incisum and C. manshurica combination exhibited synergistic effects in down-regulating TNF-α, IL-1β, PGE2 level, but not NO. Conclusions : This study suggested that combined treatment of N. incisum and C. manshurica, based on seven methods in prescription compatibility, has a synergistic effect in down-regulating inflammatory response both in vitro and in vivo models.