Ear Edema Formation Research Articles

Anti-inflammatory Activity of Sargassum micracanthum Water Extract

The anti-inflammatory effect of Sargassum micracanthum water extract (SMWE) was investigated using lipopolysaccharide (LPS)-induced inflammatory response in this study. The murine macrophage cell line RAW 264.7 cells were used and MTT assay was performed to measure the cell proliferation ability. The secretion of nitric oxide (NO), tumor necrosis factor- (TNF-), interleukin-6 (IL-6), and IL- was measured in LPS-induced RAW 264.7 cells by ELISA. The expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and nuclear transcription factor-kappa B p65 protein was studied by immunoblotting. The Balb/c mice were used for an acute toxicity test, and imprinting control region mice were purchased to evaluate a croton oil-induced ear edema. As a result, there was no cytotoxicity in the macrophage proliferation treated with SMWE compared to the control. NO levels decreased with increasing concentration of SMWE and were inhibited over 50%. Moreover, the secretion of IL-6, TNF-, and IL- was suppressed in a dose-dependent manner, especially, IL- inhibition activity was over 50% at 50 /mL. The formation of ear edema of mice was reduced at the highest dose tested compared to that in the control. Moreover, in acute toxicity test, no moralities occurred in mice administered 5,000 mg/kg body weight of SMWE over 2 weeks observation period. These results suggested that SMWE may have significant effects on inflammatory factors and be potential anti-inflammatory therapeutic materials.

Anti-inflammatory activity of methanol extract and n-hexane fraction mojabanchromanol b from Myagropsis myagroides

AimsThis study was carried out to verify the anti-inflammatory effect of methanol extract from Myagropsis myagroides (MMME) and its n-hexane fraction mojabanchromanol b. Main methodsThe murine macrophages Raw264.7 cells were used. The pro-inflammatory cytokines (IL-6, IL-1β, TNF-α) and the expression of iNOS, COX-2, and NF-κB p65 were examined by ELISA and immunoblotting. To investigate the inhibitory effect of MMME in an animal model of inflammation, an assay to determine croton oil-induced ear edema in mice was performed. Key findingsNO levels decreased with increasing concentration of MMME, and were inhibited up to 50%. The secretion of IL-6, TNF-α, and IL-1β was suppressed in a dose-dependent manner, especially at 50μg/mL, inhibition activities of cytokines were over 50%. MMME also suppressed the expression of COX-2, iNOS, and NF-κB p65, suggesting that MMME could affect the expression of inflammation related cytokines and proteins through the deregulation of NF-κB. Moreover, the formation of mouse ear edema was reduced at the highest dose tested compared to that in the control, and generated similar effects compared with prednisolone at 250mg/kg in mice ear edema evaluation test. In addition, the results in photomicrograph of mice ear tissue and mast cells also showed the same effect. After purification of fractions of MMME, it indicated that n-hexane fraction mojabanchromanol b was the most active fraction showing the inhibitory effect of IL-6 and TNF-α. SignificanceThese results suggested that MMME and mojabanchromanol b may have great effects on inflammatory factors and be potential anti-inflammatory therapeutic materials.

Intravenous anesthetic propofol suppresses prostaglandin E2 and cysteinyl leukotriene production and reduces edema formation in arachidonic acid-induced ear inflammation

Propofol is an intravenous drug widely used for anesthesia and sedation. Previously, propofol was shown to inhibit cyclo-oxygenase (COX) and 5-lipoxygenase (5-LOX) activities. Because these enzyme-inhibiting effects have only been demonstrated in vitro, this study sought to ascertain whether similar effects might also be observed in vivo. In the current studies, effects of propofol were tested in a murine model of arachidonic acid-induced ear inflammation. Specifically, propofol – as a pre-treatment -- was intraperitoneally and then topical application of arachidonic acid was performed. After 1 h, tissue biopsies were collected and tested for the presence of edema and for levels of inflammatory mediators. The results indicated that the administration of propofol significantly suppressed ear edema formation, tissue myeloperoxidase activity, and tissue production of both prostaglandin E2 and cysteinyl leukotrienes. From the data, it can be concluded that propofol could exert anti-COX and anti-5-LOX activities in an in vivo model and that these activities in turn could have, at least in part, suppressed arachidonic acid-induced edema formation in the ear.

English

The antiinflammatory activity of the methanol:methylene chloride extract of the aerial parts of Schwenckia americana L. (Solanaceae) (SAE) and its mechanisms were evaluated. Bioactivity-guided studies on SAE and its fractions was done using the egg albumin-induced rat paw edema as activity guide. Further screening for anti-inflammatory activity was accomplished using the xylene-induced ear edema and cotton pellet-induced granuloma formation. The putative anti-inflammatory mechanisms were studied using acetic acid-induced vascular permeability, carrageenan-induced neutrophil migration and heat- and hypotonic solution- induced hemolysis. Phytochemical analysis of SAE and fractions was carried out using standard procedures. Results showed that SAE and fractions significantly (P < 0.05) inhibited rat pedal edema development with the magnitude of inhibition increasing with fractionation of SAE in the order, fraction A > dichloromethane fraction > SAE. SAE also significantly (P < 0.05) inhibited acetic acid-induced vascular permeability and topical edema development in the mouse ear. Granuloma tissue formation and neutrophils count in rats, and heat and hypotonic solution- induced hemolysis of ox erythrocytes were also inhibited. The anti-inflammatory mechanism of S. americana may partly be through membrane stabilization and inhibition of vascular permeability, neutrophil migration, cellular infiltration and proliferation. Phytochemical tests revealed the presence of terpenoids and steroids in the most active fraction.   Key words: Schwenckia americana, anti-inflammatory, granuloma, ear edema, vascular permeability, neutrophil migration, membrane stabilization.

만성 및 급성 염증 모델에서 강황추출물의 효과

Anti-inflammatory effects of turmeric (Curcuma longa L.) rich in polysaccharides, as well as free of curcuminoids and turmerones were investigated in acute and chronic inflammatory models. Activity against the acute phase of inflammation was evaluated in carrageenan-induced paw edema and xylene-induced ear edema models. The results showed that turmeric extract significantly decreased paw edema volume in the first and third hours after carra- geenan injection (P≤0.05). Turmeric extract at all dose levels also significantly inhibited xylene-induced ear edema formation (P≤0.05). Activity against chronic inflammation was also evaluated in cotton pellet-induced granuloma model. Turmeric extract significantly (P≤0.05) decreased the weight of granuloma tissue on cotton pellets in a dose-dependent manner when compared to the vehicle control. Thus, the findings of the study suggest that turmeric extract in effective against both acute and chronic inflammation.

Determination of Four Volatile Compounds with Anti-Inflammatory Biological Activity in Houttuynia chordata Thunb. by Gas Chromatography and Gas Chromatography–Mass Spectrometry

An analytical procedure was developed for the determination of four volatile components with anti-inflammatory biological activity in Houttuynia cordata Thunb. by gas chromatography and gas chromatography–mass spectrometry. Four compounds (α-pinene, β-myrcene, 1-decanol and 2-undecanone) were selected as analytes because they are common major constituents in volatile compositions of Houttuynia cordata Thunb Anti-inflammatory efficacy in vivo suggested that the four compounds provided inhibitory effects on xylene-induced ear edema formation in a dose-dependent manner. The four compounds were determined by gas chromatography in mouse serum after oral administration of volatile oil and water distillate liquid of Houttuynia cordata Thunb Related chromatographic conditions were investigated and selected. A good separation of the four compounds and internal standard was achieved. The method reflected the quality of Houttuynia cordata Thunb. and its preparations.

Medicinal Potential of Phyllanthus emblica (Linn.) Fruits Extracts: Biological and Pharmacological Activities

Phyllanthus emblica (Linn.) is a common plant and fruits very popular in Bangladesh. It is a common ingredient of many traditional and herbal medicines. The intention of the present study was to explore the scientific relation with the traditional use of the fruit of Phyllanthus emblica (Linn.). Antimicrobial screening, analgesic activity, antidiarrheal activity and the brine shrimp lethality test for cytotoxic activity screening are the selected pharmacological activities. Phytochemical analysis of ethanolic fruits extract confirms that the fruit contain flavonoids, alkaloids, tannin, steroids, reducing sugar and gum. Experimental screening confirms that the fruit extract produced 19.07% and 38.67% writhing inhibition at the oral dose of 250 and 500mg/kg-body weights respectively. That means ethanol extract of the fruit of Phyllanthus emblica (Linn.) has an analgesic property. The ethanolic fruit extract of Phyllanthus emblica (Linn.) also significantly inhibited ear edema formation in xylene induced ear edema, considered as direct evidence that supporting the anti-inflammatory activity of ethanolic fruit extract of Phyllanthus emblica (Linn.) The fruit extract ha a remedy for a different bacterial disease is supported by the antibacterial screening tests. During the antidiarrheal activity screening at the dose of 500mg/kg-body weight, Phyllanthus emblica (Linn.) showed a moderate antidiarrheal activity in castor oil induced test in mice and caused an increase in latent period i.e. delayed the onset of diarrheal episode and decreased the frequency of defecation. T-test of these responses showed that the result is significant throughout the observation period. The ethanolic fruits extract have cytotoxic activity and test sample showed different mortality rate at different concentrations. The LC50 values were found to be 60µg/ml for the crude extract. The 90% mortality (LC90) values were 100µg/ml respectively. According to the results of the present investigation, we can conclude that the ethanolic fruits extract of Phyllanthus emblica (Linn.) has significant analgesic, anti-inflammatory, antimicrobial, anti-diarrheal and cytotoxic effects. This study also suggests us to isolate the active compound(s) responsible for those pharmacological properties.

A systematic review for anti-inflammatory property of clusiaceae family: a preclinical approach.

Background. Clusiaceae family (sensu lato) is extensively used in ethnomedicine for treating a number of disease conditions which include cancer, inflammation, and infection. The aim of this review is to report the pharmacological potential of plants of Clusiaceae family with the anti-inflammatory activity in animal experiments. Methods. A systematic review about experiments investigating anti-inflammatory activity of Clusiaceae family was carried out by searching bibliographic databases such as Medline, Scopus and Embase. In this update, the search terms were “anti-inflammatory agents,” “Clusiaceae,” and “animals, laboratory.” Results. A total of 255 publications with plants this family were identified. From the initial 255 studies, a total of 21 studies were selected for the final analysis. Studies with genera Allanblackia, Clusia, Garcinia or Rheedia, and Hypericum showed significant anti-inflammatory activity. The findings include a decrease of total leukocytes, a number of neutrophils, total protein concentration, granuloma formation, and paw or ear edema formation. Other interesting findings included decreased of the MPO activity, and inflammatory mediators such as NF-κB and iNOS expression, PGE2 and Il-1β levels and a decrease in chronic inflammation. Conclusion. The data reported suggests the anti-inflammatory effect potential of Clusiaceae family in animal experiments.

Antinociceptive and anti-inflammatory activities of extract and two isolated flavonoids of Carthamus tinctorius L

Ethnopharmacological relevanceSafflower (Carthamus tinctorius L.) has been long used both in the traditional system and folk medicine as an analgesic anti-inflammatory agent in China. The aim of the study was to evaluate the antinociceptive and anti-inflammatory activities of hydroalcoholic extract (HE) and two isolated kaempferol glycosides of Carthamus tinctorius L. to provide experimental evidence for its traditional use. Materials and methodsAntinociceptive effects of HE, kaempferol 3-O-rutinoside (K-3-R) and kaempferol 3-O-glucoside (K-3-G) were assessed in mice using the acetic acid-induced writhing test, formalin test and cinnamaldehyde test. The anti-inflammatory effects of HE, K-3-R and K-3-G were determined in two animal models: carrageenan-induced paw edema and xylene-induced ear edema. ResultsThe HPLC analysis showed the presence of K-3-R and K-3-G in Carthamus tinctorius L. HE (500 and 1000mg/kg) as well as K-3-R and K-3-G (150, 300 and 600mg/kg) produced significant inhibition on nociception induced by acetic acid and formalin. Oral treatment of HE, K-3-R and K-3-G at all doses significantly reduced both the nociceptive response and cinnamaldehyde-induced paw edema, effect that was superior to aspirin. In anti-inflammatory tests, HE and K-3-G significantly inhibited the paw edema during the both phases of carrageenan-induced inflammation while K-3-G suppressed the late phase inflammation only. HE (400 and 800mg/kg) and K-3-G (200, 400, 800mg/kg) produced significant dose-dependent inhibition of xylene-induced ear edema development. K-3-R only suppressed ear edema formation at a high dose (800mg/kg). ConclusionsThese results demonstrate that Carthamus tinctorius L. extract possess remarkable antinociceptive and anti-inflammatory activities which may be due to K-3-R and K-3-G at least in part, supporting the folkloric usage of the plant to treat various inflammatory and pain diseases.

Erucin Exerts Anti-Inflammatory Properties in Murine Macrophages and Mouse Skin: Possible Mediation through the Inhibition of NFκB Signaling

Erucin, an isothiocyanate, is a hydrolysis product of glucoerucin found in arugula and has recently been reported to have anti-cancer properties in various cancer cells. In this study, we assessed the anti-inflammatory effects of erucin and the underlying mechanisms, using lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophages and 12-O-tetradecanoylphorbol-13-acetate-treated mouse skin. In RAW 264.7 cells, erucin (2.5, 5 μmol/L) inhibited LPS-induced production of nitric oxide and prostaglandin E2. Erucin inhibited LPS-induced degradation of the inhibitor of κBα and translocation of p65 to the nucleus and, subsequently, reduced LPS-induced nuclear factor κB (NFκB) DNA binding activities, as well as the transcriptional activity of NFκB, leading to the decreased expression of NFκB-target genes, including tumor necrosis factor-α, interleukin (IL)-6, IL-1β, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2, as well as transcriptional activity of iNOS and COX-2. In mice, erucin (100, 300 nmoles) treatment significantly inhibited phorbol ester-induced formation of ear edema and expression of iNOS and COX-2 proteins. These results indicate that erucin exerts a potent anti-inflammatory activity by inhibiting the pro-inflammatory enzymes and cytokines, which may be mediated, at least in part, via the inhibition of NFκB signaling.

Antimicrobial, anti-inflammatory activities and toxicology of phenylethanoid glycosides from Monochasma savatieri Franch. ex Maxim

Ethnopharmacological relevanceMonochasma savatieri Franch. ex Maxim is used for treating many diseases in Traditional Chinese Medicine. Aim of the studyThe present study was designed to evaluate antibacterial, anti-inflammatory activities and toxicity of the total phenylethanoid glycosides from Monochasma savatieri Franch. ex Maxim (TPG). Materials and methodsThe antibacterial activity of TPG was checked by MIC and MBC in vitro; survival of mice with Pseudomonas aeruginosa or Staphylococcus aureus infection-induced sepsis was investigated to evaluate antibacterial activity of TPG in vivo. Additionally, antibacterial activities of TPG were also investigated in a Pseudomonas aeruginosa infection-induced pneumonia in the mice model. Cotton pellet induced granuloma and xylene induced ear swelling in mice models were used to quantify the anti-inflammatory activity. ResultsTPG showed a significant possess bacteriostatic properties against five bacteria strains at a concentration between 0.0625 and 16mg/ml. Moreover, TPG has bactericidal activity against Pseudomonas aeruginosa, Streptococcus pneumoniae or Escherichia coli. TPG (60, 120, and 180mg/kg) prolonged survival rate of mice with Pseudomonas aeruginosa or Staphylococcus aureus infection-induced sepsis. In addition, TPG (180mg/kg) could reduce the bacterial colony-forming units in lung tissue. Furthermore, TPG (60–180mg/kg) had significantly reduced xylene-induced ear edema and granulomat formation induced by cotton pellet at a dose-dependent manner. In addition, administration of TPG (1.5g/kg) for 15 days did not result in toxicities in liver, kidney, spleen and thymus tissue in rats. ConclusionThese results indicated that TPG might be useful for the development of a novel treatment for respiratory infections or pneumonia caused by Pseudomonas aeruginosa or Staphylococcus aureus.

Mechanisms by Which Licochalcone E Exhibits Potent Anti-Inflammatory Properties: Studies with Phorbol Ester-Treated Mouse Skin and Lipopolysaccharide-Stimulated Murine Macrophages

In this study we found that licochalcone E (LicE), a recently isolated retrochalcone from Glycyrrhiza inflata, exhibits potent anti-inflammatory effects in 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse ear edema and lipopolysaccharide (LPS)-stimulated RAW 264.7 murine macrophage models. Topical application of LicE (0.5–2 mg) effectively inhibited TPA-induced (1) ear edema formation; (2) phosphorylation of stress-activated protein kinase/c-Jun-N-terminal kinase (SAPK/JNK), c-Jun, and extracellular signal regulated kinase 1/2; and (3) expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 proteins in mouse skin. The treatment of RAW 264.7 cells with LicE (2.5–7.5 μmol/L) induced a profound reduction in LPS-induced (1) release of NO and prostaglandin E2; (2) mRNA expression and secretion of interleukin (IL)-6, IL-1β and tumor necrosis factor-α; (3) promoter activity of iNOS and COX-2 and expression of their corresponding mRNAs and proteins; (4) activation of AKT, p38 mitogen activated protein kinase (MAPK), SAPK/JNK and c-Jun; (5) phosphorylation of inhibitor of κB (IκB) kinase-αβ and IκBα, degradation of IκBα, translocation of p65 (RelA) to the nucleus and transcriptional activity of nuclear factor (NF)-κB; and (6) transcriptional activity of activator protein (AP)-1. These results indicate that the LicE inhibition of NF-κB and AP-1 transcriptional activity through the inhibition of AKT and MAPK activation contributes to decreases in the expression of pro-inflammatory cytokines and the inducible enzymes iNOS and COX-2.

Evaluation of anti-inflammatory and antinociceptive activity of Triphala recipe

The anti-inflammatory and antinociceptive activities of Triphala recipe were studied in animal models. Triphala recipe (4 mg/ear) significantly exhibited an inhibitory effect on the ear edema formation induced by ethyl phenylpropiolate-induced, but not on the arachidonic acid-induced ear edema in rats. Furthermore, Triphala recipe at the doses of 300, 600 and 1,200 mg/kg significantly reduced carrageenan-induced hind paw edema. Next, the anti-inflammatory action in chronic inflammation was measured using the cotton pellet-induced granuloma formation assay in rats. Triphala recipe (1,200 mg/kg) reduced neither transudative weight nor granuloma formation. It also did not affect on body weight gain and thymus weight indicating that Triphala recipe does not have a steroid-like effect. In antinociceptive study, Triphala recipe (300, 600, 1,200 mg/kg), elicited significant inhibitory effect on both phases, especially in late phase, of the formalin test in mice suggesting that the antinociceptive action of Triphala recipe may be via both peripheral and at least partly centrally acting.

Analgesic, anti-inflammatory and anti-diarrheal activities of ethanolic leaf extract of Typhonium trilobatum L. Schott

ObjectiveTo explore the efficacy of ethanolic leaf extract of Typhonium trilobatum L. Schott in treating diarrhea, pain and inflammation using experimental models. MethodsIn the present study, acetic acid-induced writhing, xylene-induced ear edema and castor oil-induced diarrheal model were used to evaluate the analgesic, anti-inflammatory and anti-diarrheal activities, respectively. Acute toxicity test was carried out to fix the safe doses of the plant extract. ResultsThe plant extract demonstrated a significant inhibition of writhing (P < 0.01) compared with the control group in acetic acid-induced writhing test in mice. The extract also significantly inhibited the xylene induced ear edema formation (P < 0.05). In anti-diarrheal test, the extract significantly decreased the frequency of defecation and increased the mean latent period (P < 0.01) in castor oil-induced diarrheal model mice at the doses of 250 and 500 mg/kg body weight. ConclusionsThese results suggest that the extract possesses significant analgesic, anti-inflammatory and anti-diarrheal activities that support to the ethnopharmacological uses of this plant.

In vivo participation of nitric oxide in hyperproliferative epidermal phenomena in mice

A significant involvement of nitric oxide (NO) in the process of keratinocyte proliferation is reported with many divergences. To determine the involvement of NO in the hyperproliferative process of epidermis in vivo, non-selective inhibitor (NG-nitro-l-arginine-methyl ester.HCl: L-NAME) and selective inhibitors for inducible NO synthase (iNOS) and neuronal NO synthase (nNOS) (Aminoguanidine: AG and 7-Nitroindazole: 7-NI, respectively) and a NO-donor (Sodium nitroprusside: SNP) were topically applied twice a day in mice ear treated with multiple applications of croton oil. L-NAME and 7-NI treatments decreased and SNP increased ear edema formation. However, ear weight was reduced in groups that received L-NAME and 7-NI, while the AG and SNP groups presented an increment. The histological evaluation of epidermis thickness showed that all NOS inhibitors were able to prevent the increase in epidermis width caused by croton oil, while SNP contributed to enlargement. The same results were observed in the PCNA staining, where treatments with NOS inhibitors caused a reduction in the number of cells in the epidermis, while SNP caused an enhancement. 7-NI treatment reduced polymorphonuclear and mononuclear leukocytes migration when compared to the control group. The AG application increased the migration of polymorphonuclear and mononuclear cells, while the SNP enhanced only the polymorphonuclear cells. Therefore, in the skin NO produced by nNOS is involved in the control of keratinocyte hyperproliferation, with the contribution of iNOS. In the animal model of cutaneous chronic inflammation by croton oil, NO is involved in the exudation and leukocyte migration, with participation of all three enzymes.

Antinociceptive, anti-inflammatory and antidiarrheal activities of ethanolic calyx extract of Hibiscus sabdariffa Linn. (Malvaceae) in mice

To evaluate the antinociceptive, anti-inflammatory and antidiarrheal activities of the ethanolic calyx extract of Hibiscus sabdariffa Linn. in mice. In the present study, the dried calyxes of H. sabdariffa were subjected to extraction with 95% ethanol and the extract was used to investigate the possible activities. Antinociceptive activity of the extract was evaluated by using the acetic acid-induced writhing test. The anti-inflammatory effect of the extract was tested by using the xylene-induced ear edema model mice. Castor oil-induced diarrheal model mice were used to evaluate the antidiarrheal activity of the extract. In acetic acid-induced writhing test, the extract produced inhibited writhing in mice significantly compared with the blank control (P<0.01). The extract showed significant inhibition of ear edema formation in xylene-induced ear edema model mice in a dose-related manner compared with the blank control (P<0.01). The extract demonstrated a significant antidiarrheal activity against castor oil-induced diarrheal in mice in which it decreased the frequency of defecation and increased the mean latent period at the doses of 250 and 500 mg/kg body weight (P<0.01). The above mentioned findings indicate that the calyx extract of H. sabdariffa possesses significant antinociceptive, anti-inflammatory and antidiarrheal activities that support its uses in traditional medicine.

Anti-inflammatory activity of Sorbus commixta water extract and its molecular inhibitory mechanism

Sorbus commixta Hedl. (Rosaceae) is a well known traditionally valuable medicinal plant in Korea, China and Japan. This plant has been prescribed for long time for various inflammatory symptoms such as asthma, bronchitis, gastritis and dropsy. Although a number of pharmacological properties have already been demonstrated, the anti-inflammatory effect of this plant and its associated molecular mechanisms has not yet been fully investigated. In order to address the anti-inflammatory activity of S. commixta water extract (Sc-WE), lipopolysaccharide (LPS)-stimulated macrophages were employed and production of inflammatory mediators by these cells were evaluated. Sc-WE significantly suppressed the production of nitric oxide (NO) and prostaglandin (PG)E(2) in a dose-dependent manner and blocked ear edema formation induced by arachidonic acid in mouse. In addition, this extract effectively diminished the mRNA levels of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2, indicating that the inhibition occurs at the transcriptional level. Interestingly, Sc-WE remarkably blocked NF-κB translocation and its upstream signaling events by inhibition of κBα (IκBα), IκBα kinase (IKK), Akt (protein kinase B), phosphoinositide-dependent kinase 1 (PDK1), p85/phosphoinositide-3-kinase (PI3K), as per the results obtained from the reporter gene assay and immunoblotting analysis. More intriguingly, Sc-WE suppressed activities of Src and Syk kinases as well as their phosphorylation levels without altering molecular complex formation between them and toll like receptor (TLR)4 or MyD88, an adaptor protein of TLR4-mediated signaling. Therefore, our results suggest that Sc-WE can be developed as a potent anti-inflammatory remedy, acting by suppressing the inflammatory signaling cascade composed of Src, Syk, and NF-κB.

3,3′‐diindolylmethane suppresses 12‐O‐tetradecanoylphorbol‐13‐acetate‐induced inflammation and tumor promotion in mouse skin via the downregulation of inflammatory mediators

3,3'-Diindolylmethane (DIM) is a major acid-condensation product of indole-3-carbinol and is present in cruciferous vegetables. In this study, we evaluated the effects of DIM on antiinflammatory and antitumor promotion activity in mouse skin and explored the relevant mechanisms. When 12-O-tetradecanoylphorbol-13-acetate (TPA) was applied topically to the mouse ear to induce inflammation, DIM pretreatment effectively inhibited TPA-induced ear edema formation. To evaluate the mechanisms underlying DIM's antiinflammatory effects, DIM was topically treated to the shaved backs of mice 30 min before TPA treatment. DIM inhibited the TPA-induced increases in the expression of cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS), chemokine (C-X-C motif) ligand (CXCL) 5, and interleukin (IL)-6 in mouse skin. DIM also inhibited nuclear factor-kappa B (NF-kappaB)'s DNA binding activity, the nuclear translocation of p65, and the degradation of inhibitor of kappaB (IkappaB) alpha in TPA-stimulated mouse skin. Furthermore, DIM reduced TPA-induced increases in the activity of extracellular signal regulated protein kinase (ERK)-1/2 and IkappaB kinase (IKK). When mouse skin papillomas were initiated via the topical application of 7,12-dimethylbenz[alpha]anthracene (DMBA) and promoted with repeated topical applications of TPA, repeated topical applications of DIM prior to each TPA treatment significantly suppressed the incidence and multiplicity of the papillomas. DIM also reduced the expression of COX-2 and iNOS, ERK phosphorylation, and the nuclear translocation of p65 in papillomas. Collectively, these results show that DIM exerts antiinflammatory and chemopreventive effects in mouse skin via the downregulation of COX-2, iNOS, CXCL5, and IL-6 expression, which may be mediated by reductions in NF-kappaB activation.

Pharmacological evaluation of Alstonia scholaris: Anti-inflammatory and analgesic effects

Ethnopharmacological relevance Alstonia scholaris (Apocynaceae) has been historically used in “dai” ethnopharmacy to treat chronic respiratory diseases. The leaf extract, developed as a commercially available traditional Chinese medicine, used to release tracheitis and cold symptom, has also been prescribed in hospitals and sold over the counter in drug stores. Aim of the study The investigation evaluated the anti-inflammatory and analgesic activities of the ethanolic extract, fractions and main alkaloids of Alstonia scholaris leaf to provide experimental evidence for its traditional and modern clinical use. Besides, to discover the active fraction and components for further better use in Chinese medicine is hopeful. Materials and methods The leaf of Alstonia scholaris was extracted with ethanol and then separated into different fractions. Furthermore, alkaloids were isolated by phytochemical method. The analgesic activities were investigated using acetic acid-induced writhing, hot-plate and formalin tests in mice. The anti-inflammatory activities were carried out in vivo and in vitro, including xylene-induced ear edema and carrageenan-induced air pouch formation in mice, and COX-1, -2 and 5-LOX inhibition. Results It has been exhibited that the EtOAc and alkaloid fractions reduced acetic acid-induced writhing response in mice, significantly. The ethanolic extract, EtOAc and alkaloid fractions remarkably inhibited xylene-induced ear edema. Further investigation was focused on the alkaloids fraction and three main alkaloids isolated from the alkaloids fraction, in different animal models. Alkaloids reduced acetic acid-induced writhing response, and xylene-induced ear edema in mice. In the hot-plate test, alkaloids did not increase the latency period of mice obviously. In the formalin test, alkaloids did not inhibit the licking time in first phase, but significantly inhibited the licking time in second phase of mice. Alkaloids increased significantly SOD activity and decreased levels of NO, PGE2 and MDA significantly, in air pouch mice model. Moreover, some alkaloids isolated from the leaf of Alstonia scholaris exhibited inhibition of COX-1, COX-2 and 5-LOX in vitro anti-inflammatory assay, which supported alkaloids as the bioactive fraction. Conclusions The alkaloids fraction of Alstonia scholaris leaf, three main alkaloids, picrinine, vallesamine and scholaricine, may produce the anti-inflammatory and analgesic effect peripherally based on several in vivo assays. In in vitro tests, alkaloids exhibited inhibition of inflammatory mediators (COX-1, COX-2 and 5-LOX), which is accordant with results on animal models. Besides, COX-2/5-LOX dual inhibitors found in the experiment, such as 16-formyl-5α-methoxystrictamine, picralinal, and tubotaiwine might be valuable for further attention.

Benzyl isothiocyanate exhibits anti-inflammatory effects in murine macrophages and in mouse skin

Benzyl isothiocyanate (BITC) is detected in abundance in Brassica vegetables, and some previous studies have demonstrated that BITC may potentially function as a chemopreventive agent in humans. This study examined whether BITC inhibits lipopolysaccharide (LPS)-induced inflammatory responses in Raw 264.7 macrophages and 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced ear edema formation. The treatment of macrophages with various concentrations of BITC resulted in a dose-dependent reduction in the LPS-induced secretion of interleukin (IL)-1beta, TNF-alpha, and IL-6 and their corresponding mRNA levels, as well as in the production of nitric oxide and PGE(2). Consistent with these findings, BITC inhibited the LPS-induced expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 proteins and their corresponding mRNAs. BITC inhibited LPS-induced phosphorylation and the degradation of the inhibitor of kappaBalpha, translocation of p65 into the nucleus, and the DNA binding activity and transcriptional activity of NFkappaB. Moreover, the LPS-stimulated phosphorylation of extracellular signal regulated kinase (ERK)1/2 and Akt was suppressed by BITC. BITC also inhibited ear edema formation and the protein expression of iNOS and COX-2 in mouse skin treated with TPA. We demonstrate that BITC is a potent anti-inflammatory agent, and the anti-inflammatory properties of BITC may result from the downregulation of NFkappaB signaling.