ABSTRACT
OBJECTIVE: Despite some knowledge gaps in scientific evidence, MgCl2 is largely used for pain relief in musculoskeletal diseases. Mg salts were shown to provide analgesia postoperatively in orthopedic surgery and low Mg levels were linked to arthritis development and severity. We determined the anti-inflammatory activity of MgCl2 in an acute arthritis model. METHODS: Mice received 0.1 mg/25µL Zymosan (Zy) or saline into the knees. Joint pain was evaluated using von Frey test; cell influx, and interleukin (IL)-1 level were assessed in joint lavage at 6 h. Synovia were excised for histopathology and analysis of immunoexpression of nuclear factor kappa B (NFκB) and tumor necrosis factor (TNF)-α. Groups (n = 6/group) received either 90 mg/kg MgCl2/100 µL or saline per os (systemic) or 500 µg/25 µL MgCl2 or saline intra-articularly (i.a.) 30 min prior to Zy. RESULTS: MgCl2 given either systemically or locally significantly reduced cell influx (p = 0.0012 and p = 0.0269, respectively), pain (p = 0.0005 and p = 0.0038, respectively), and intra-articular IL-1 level (p = 0.0391), as compared to saline. Systemic MgCl2 significantly decreased NFκB (p < 0.05) immmunoexpression, as compared to saline. CONCLUSION: MgCl2 given systemically or locally displayed anti-inflammatory activity in a severe acute arthritis model reducing cell influx, pain, and cytokine release. MgCl2 operates at least partially via inhibiting NFκB activation. This is the first in vivo demonstration that MgCl2 decreases cytokine release in arthritis, prompting reduction of inflammation and pain relief.
Subject(s)
Arthritis, Experimental , Rats , Humans , Mice , Animals , Magnesium Chloride/therapeutic use , Rats, Wistar , Arthritis, Experimental/drug therapy , Cytokines , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/therapeutic use , Tumor Necrosis Factor-alpha , Interleukin-1 , PainABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: The cause of irritable bowel syndrome (IBS), a functional gastrointestinal (GI) disorder, remains unclear. Banhasasim-tang (BHSST), a traditional herbal medicines mixture, mainly used to treat GI-related diseases, may have a potential in IBS treatment. IBS is characterized by abdominal pain as the main clinical symptom, which seriously affects the quality of life. AIM OF THE STUDY: We conducted a study to evaluate the effectiveness of BHSST and its mechanisms of action in treating IBS. MATERIALS AND METHODS: We evaluated the efficacy of BHSST in a zymosan-induced diarrhea-predominant animal model of IBS. Electrophysiological methods were used to confirm modulation of transient receptor potential (TRP) and voltage-gated Na+ (NaV) ion channels, which are associated mechanisms of action. RESULTS: Oral administration of BHSST decreased colon length, increased stool scores, and increased colon weight. Weight loss was also minimized without affecting food intake. In mice administered with BHSST, the mucosal thickness was suppressed, making it similar to that of normal mice, and the degree of tumor necrosis factor-α was severely reduced. These effects were similar to those of the anti-inflammatory drug-sulfasalazine-and antidepressant-amitriptyline. Moreover, pain-related behaviors were substantially reduced. Additionally, BHSST inhibited TRPA1, NaV1.5, and NaV1.7 ion channels associated with IBS-mediated visceral hypersensitivity. CONCLUSIONS: In summary, the findings suggest that BHSST has potential beneficial effects on IBS and diarrhea through the modulation of ion channels.
Subject(s)
Irritable Bowel Syndrome , Plants, Medicinal , Mice , Animals , Irritable Bowel Syndrome/drug therapy , Irritable Bowel Syndrome/chemically induced , Quality of Life , Diarrhea/chemically induced , Diarrhea/drug therapy , Plant Extracts/pharmacology , Plant Extracts/therapeutic use , TRPA1 Cation ChannelABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Viridiflorol was identified and isolated from the essential oil of Allophylus edulis leaves (EOAE). A. edulis was used as "tereré", which is a drink made by the infusion of herbs in cold water, to treat pain (toothache and headache). All anti-nociceptive (analgesic) and anti-arthritic properties of EOAE and viridiflorol have not been completely scientifically clarified. AIM OF THE STUDY: The aim of the present study was to investigate the analgesic (anti-hyperalgesic and anti-nociceptive) and anti-arthritic properties of EOAE and viridiflorol using in vivo models. MATERIALS AND METHODS: The oral administration (p.o.) of EOAE (30, 100 and 300 mg/kg), viridiflorol (30, 100 and 200 mg/kg), morphine (1 mg/kg, subcutaneous route (s.c.)) and the intraplantar (local) administration (i.pl.) of viridiflorol (100 µg/paw) were tested using formalin model in Swiss mice. EOAE (100 mg/kg, p.o.), viridiflorol (200 mg/kg, p.o.), and dexamethasone (1 mg/kg, s.c.) were tested by zymosan-articular inflammation and in open-field models. Viridiflorol (0.3, 20 and 200 µg/paw) was also tested in carrageenan model, and viridiflorol (200 µg/paw) was also tested in tumor necrosis factor-α (TNF-α), and dopamine (DOPA) models. RESULTS: The oral administration of EOAE (100 and 300 mg/kg, p.o.), viridiflorol (200 mg/kg, p.o.), morphine (1 mg/kg, s.c.) (MOR) and local administration of viridiflorol (100 µg/paw) significantly inhibited edema and nociception in formalin model. Oral treatments with EOAE and viridiflorol (200 mg/kg) did not cause motor impairment in the open field test since they did not reduce locomotor activity. EOAE, viridiflorol and dexamethasone significantly reduced mechanical hyperalgesia, edema, total leukocytes, polymorphonuclear cells, nitric oxide and protein exudation in the zymosan-induced articular inflammation model. The local administration of viridiflorol (200 µg/paw, i.pl.) significantly inhibited mechanical hyperalgesia and edema induced by carrageenan, TNF-α and DOPA. CONCLUSIONS: This study confirms the potential anti-arthritic, anti-nocicepttive and anti-hyperalgesic properties of EOAE and viridiflorol. These properties could explain, at least in part, the folk use of A. edulis against including pain (toothache and headache). Viridiflorol could be partially responsible for the EOAE anti-hyperalgesic, anti-nociceptive and anti-arthritic properties and its mechanism of action could involve the inhibition of TNF-α and DOPA pathways.
Subject(s)
Oils, Volatile , Animals , Mice , Analgesics , Anti-Inflammatory Agents , Carrageenan , Dexamethasone/therapeutic use , Dihydroxyphenylalanine , Disease Models, Animal , Edema/chemically induced , Edema/drug therapy , Formaldehyde , Headache/drug therapy , Hyperalgesia/chemically induced , Hyperalgesia/drug therapy , Inflammation/drug therapy , Morphine Derivatives , Plant Extracts/pharmacology , Plant Extracts/therapeutic use , Toothache/drug therapy , Tumor Necrosis Factor-alpha , ZymosanABSTRACT
Calcipotriol, a vitamin D analogue, is an antiproliferative and anti-inflammatory drug currently used in psoriasis. Here, our aim was to analyse the safety of calcipotriol for cartilage and bone in alleviated-dose (0.1 mg instead of usual ≥1mg dose) zymosan-induced arthritis in rats. Theoretically, high doses of vitamin D or analogues could have detrimental effects on bone or cartilage. The rats were divided into four groups: vehicle (n = 9), dexamethasone 0.1 mg/kg (n = 9), calcipotriol 0.1 mg/kg (n = 8) and negative control (n = 10) with no injections. Arthritic rats were given phosphate-buffered saline (PBS) injections to left knees as a control. After euthanasia on day 8, all knees were imaged with micro-computed tomography for surface lesions and decalcified for histological analyses. Contrary to our expectations, no significant changes could be observed in the tomography data and histological scores among the three treatment groups or between the vehicle-treated and non-arthritic group. Calcipotriol did not cause adverse effects on cartilage or subchondral bone within a week, suggesting that it could be safely used in local treatment of arthritis. The alleviated model caused synovitis with local and systemic inflammatory response without cartilage erosions, which might be useful in studying self-limiting synovitis where cartilage or bone effects are not of primary interest.
Subject(s)
Arthritis, Experimental , Cartilage, Articular , Synovitis , Rats , Animals , Zymosan/adverse effects , Vitamin D , Rats, Wistar , Arthritis, Experimental/chemically induced , Arthritis, Experimental/drug therapy , X-Ray Microtomography , Cartilage, Articular/pathology , Synovitis/chemically induced , Synovitis/pathologyABSTRACT
Matrix metalloproteinases (MMPs) play a crucial role in the degenerative course of rheumatic disorders. They are responsible for cartilage and other joint-associated tissues breakdown. Amid arthritis treatments, photobiostimulation (PBM), a non-thermal and non-invasive low-power laser application, appears to be an outstanding therapy alternative once it has succeeded in MMPs modulation. Thus, this study aimed to evaluate the PBM effects of low infrared laser (830 nm), testing two different energy densities (3 and 30 Jcm-2) in MMP-2, MMP-9, MMP-13, and MMP-14 as well as the inhibitor TIMP-2 expressions using zymosan-induced arthritis model. C57BL/6 mice were distributed into four groups (n = 8): zymosan-induced arthritis without treatment; zymosan-induced arthritis and dexamethasone-treated; zymosan-induced arthritis and PBM at energy density of 3 Jcm-2 treated; and zymosan-induced arthritis and PBM at energy density of 30 Jcm-2 treated. MMPs and TIMP-2 mRNA relative levels by qRT-PCR and proteins expression by immunohistochemical and Western blotting techniques were performed after PBM treatment in the inflamed joint. Our results demonstrated PBM could modulate both mRNA relative levels and proteins expression of the MMP-2, -9, -13, -14, and TIMP-2 in joint tissues, decreasing MMP-9 protein expression and increasing TIMP-2 protein expression. PBM promotes a better arthritis prognostic, modulating metalloproteinase and its inhibitor, especially MMP-9 and TIMP-2 protein expression that is important inflammatory markers. These findings may also corroborate that PBM may regulate MMPs expression using different pathways.
Subject(s)
Arthritis , Low-Level Light Therapy , Animals , Mice , Arthritis/chemically induced , Arthritis/genetics , Arthritis/radiotherapy , Matrix Metalloproteinases/genetics , Matrix Metalloproteinases/metabolism , Mice, Inbred C57BL , RNA, Messenger/genetics , Tissue Inhibitor of Metalloproteinase-1/genetics , Tissue Inhibitor of Metalloproteinase-1/metabolism , Tissue Inhibitor of Metalloproteinase-2/genetics , Tissue Inhibitor of Metalloproteinase-2/metabolism , ZymosanABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Geranium sanguineum L. is used for treatment of inflammations, anemia, malignant diseases of the blood-forming organs, diarrhea, respiratory infections, etc. Only flavonoids in root extracts have been elucidated as immunostimulating and anti-inflammatory compounds, and polysaccharides in the herb have not been examined. AIM OF THE STUDY: to compare the chemical features of polysaccharide complexes (PSCs) from leaves (GSL-PSC) and roots (GSR-PSC) of G. sanguineum, as well as their immunomodulatory activities on leukocytes after inflammation, and effects on the growth of different bacteria. MATERIALS AND METHODS: The samples were isolated by water extraction and their structural features were studied by 2D NMR spectroscopy. The stimulatory effects of both PSCs on human leukocytes were analyzed with flow cytometry. Their suppressive activities on the oxidative burst in blood and derived neutrophils against opsonized zymosan and phorbol myristate acetate were investigated. The effects of the samples on viability, NO and interleukin 6 (IL-6) syntheses in RAW264.7 cells after inflammation with lipopolysaccharides (LPS) were tested. The prebiotic and anti-biofilm activities of the PSCs were evaluated. RESULTS: The total carbohydrate content in the samples was significant (73.6-76.8%). GSL-PSC contained pectins, which were rich in homogalacturonan (HG), and smaller amounts of rhamnogalacturonan (RG) type I, decorated by 1,5-α-L-Araf, 1,4- and 1,6-ß-D-Galp chains. GSR-PSC contained starch, followed by pectins with lower HG content and more RG-I regions, substituted by 1 â 3,5-α-L-arabinans and 1 â 3,6-ß-D-galactans. GSL-PSC and GSR-PSC (200 µg/mL) increased monocyte and granulocyte cell counts, but GSR-PSC also elevated T helper and B cell levels in a normal and activated state. GSR-PSC triggered a dose-dependent (50-200 µg/mL) oxidative burst in blood, but alleviated it after inflammation even in blood-derived neutrophils. It was free of LPS, and activated NO and IL-6 productions in RAW264.7 cells better than GSL-PSC, without affecting their viability. Both PSCs (2.0%, w/v) stimulated probiotic co-cultures between Clostridium beijerinckii strains and Lactobacillus sp. ZK9, and inhibited the growth and biofilm formation of Escherichia coli, Streptococcus mutans and Salmonella enterica. CONCLUSIONS: The PSs in G. sanguineum could be involved in the stimulatory effects on blood-forming organs and anti-inflammatory action of aqueous root extracts in case of infections. These PSs should be included in synbiotic foods to support the treatment of inflammations and infections in the gut.
Subject(s)
Geranium , Polysaccharides , Animals , Anti-Inflammatory Agents , Geranium/chemistry , Humans , Inflammation/drug therapy , Interleukin-6 , Lipopolysaccharides , Mice , Pectins/pharmacology , Polysaccharides/pharmacology , RAW 264.7 CellsABSTRACT
Six flavonoids were identified and isolated from the ethanolic extract of Alternanthera tenella Colla (Amaranthaceae) whole plant (EEAT) including 2â³-O-ß-D-glucopyranosyl-vitexin (A19). Flavonoids have anti-inflammatory activity; however, the 2â³-O-ß-D-glucopyranosyl-vitexin anti-inflammatory property was not totally explored. The aim of the present study was to investigate the anti-inflammatory effects of ethanolic extract from A. tenella whole plant and isolated flavone C-glycoside A19 in models of inflammation. The oral administration (p.o.) of EEAT (30, 100, and 300 mg/kg), A19 (0.1, 1, and 10 mg/kg), and prednisolone (3 mg/kg) were tested against the carrageenan-induced paw edema in Swiss mice. The EEAT (100 mg/kg, p.o.), A19 (1 mg/kg, p.o.), and prednisolone (3 mg/kg, p.o.) were tested in the zymosan-articular inflammation, carrageenan-pleurisy, and complete Freund's adjuvant (CFA) models in Swiss mice. In silico analysis and search for structural relationships between vitexin derivatives flavones present in the EEAT and TNF-α inhibitors were performed. EEAT, A19, and prednisolone significantly inhibited (i) edema, mechanical hyperalgesia in carrageenan-induced paw inflammation; (ii) leukocyte migration and protein extravasation in carrageenan-induced pleurisy; (iii) knee edema, mechanical hyperalgesia, and leukocyte migration in articular inflammation induced by zymosan. Still the CFA induced the increase in myeloperoxidase and N-acetylglucosaminidase activities, EEAT, A19, and prednisolone significantly inhibited these enzymes. The in silico analysis showed that molecules with similar structure to apigenin and vitexin have a potential inhibition on the TNF system. This study confirms the anti-inflammatory properties of EEAT and A19. The C-glycosylated flavone A19 could be responsible for the EEAT anti-edematogenic and anti-hyperalgesic effects and a potential TNF-α inhibitor.
Subject(s)
Amaranthaceae , Anti-Inflammatory Agents/therapeutic use , Ethanol/therapeutic use , Flavones/therapeutic use , Plant Extracts/therapeutic use , Animals , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/isolation & purification , Dose-Response Relationship, Drug , Edema/drug therapy , Edema/pathology , Female , Flavones/chemistry , Flavones/isolation & purification , Male , Mice , Mice, Inbred C57BL , Plant Components, Aerial , Plant Extracts/chemistry , Plant Extracts/isolation & purificationABSTRACT
The aim of this study was to explore the effects of ethanol extracts from Portulaca oleracea L. (ePO) on joint inflammation and to explain the underlying mechanisms. A joint inflammation mouse model was constructed by injecting zymosan, and the Von Frey method was employed and the joint thickness measured. The numbers of leukocytes, neutrophils, and monocytes were counted in the joint cavity and the infiltration of inflammatory cells was assessed by joint histopathological analysis. The mRNA levels of inflammatory cytokines were determined by quantitative RT-PCR and their secretion levels were determined by specific ELISAs. Pre-treatment with ePO inhibited articular mechanical hyperalgesia and edema and ameliorated the recruitment of mononuclear neutrophils and leukocytes. In addition, pre-treatment with ePO improved pathological alternations in the joint tissues by reducing the number of inflammatory cells. Pre-treatment with ePO regulated the nuclear factor erythroid 2-related factor 2 (Nrf2)-related proteins and thereby inhibited oxidative stress. In addition, ePO inhibited NLR family pyrin domain containing 3 (NLRP3) inflammasome-related genes (NLRP3, ASC, pro-caspase-1 and pro-IL-1ß), modulated inflammatory cytokines and the activation of NF-κB. ePO attenuated zymosan-induced joint inflammation by regulating oxidative stress, NLRP3 inflammasome, and NF-κB.
Subject(s)
Analgesics/therapeutic use , Arthritis/drug therapy , NF-E2-Related Factor 2/metabolism , Nociceptors/physiology , Oxidative Stress/drug effects , Plant Extracts/therapeutic use , Analgesics/chemistry , Animals , Cytokines/metabolism , Disease Models, Animal , Down-Regulation , Ethanol/chemistry , Humans , Inflammation Mediators/metabolism , Male , Mice , NF-E2-Related Factor 2/genetics , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Nociceptors/drug effects , Plant Extracts/chemistry , Portulaca/immunology , ZymosanABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Stem barks of Caesalpinia ferrea Mart. Ex Tul. (Caesalpiniaceae), also known as pau-ferro jucá or jucaína, are popularly used to treat contusions, diabetes, rheumatism and other inflammatory conditions in the form of tea, lick or decoction. OBJECTIVE: To evaluate the effect of the polysaccharide-rich extract obtained from C. ferrea stem barks (PE-Cf) in mice models of acute inflammation induced by zymosan and the involvement of oxidative stress biomarkers. MATERIALS AND METHODS: Mice were treated with PE-Cf (0.001, 0.01, 0.1, 1 mg/kg) by endovenous route (i.v.) or per oral (p.o.) 30 or 60 min before injection of the inflammatory stimuli zymosan (0.5 mg; intraperitoneal or subcutaneous intraplantar). The inflammatory parameters (edema, nociception, leukocyte migration) and oxidative stress markers (myeloperoxidase-MPO, malondialdehyde-MDA, nitrite, reduced glutathione-GSH, glutathione peroxidase-GPx) were evaluated in the models of paw edema (hidropletysmometry/expressed as ml or area under curve-AUC) and peritonitis (optical microscopy/expressed as n° of cells/mm3 of peritoneal fluid). Statistical analysis was performed by ANOVA, followed by Bonferroni test. RESULTS: PE-Cf (0.1, 0.01 and 1 mg/kg) dose-dependently inhibited paw edema, showing maximal effect (74%) at 1 mg/kg in the 5th (52 ± 9.6 µl vs. zymosan: 204 ± 3.6 µl). PE-Cf (1 mg/kg) also inhibited by 43% MPO activity in the paw tissues (17 ± 1 vs. zymosan: 30 ± 2.6 U/mg). Besides, 4 h after peritonitis induction, PE-Cf (1 mg/kg) reduced neutrophil migration by 84% (432 ± 45 vs. zymosan: 2651 ± 643 cells/mm3); visceral nociception by 76% (3 ± 0.6 vs. zymosan: 16 ± 4 writhes); nitric oxide by 73% (0.131 ± 0.033 vs. zymosan: 0.578 ± 0.185 NO2-/NO3-ml); MDA (98 ± 10 vs. zymosan:156 ± 21 U/ml), and increased GSH by 65% (736 ± 65 vs. zymosan: 259 ± 58 µmol/ml) and GPx by 72% (0.037 ± 0.007 vs. zymosan: 0.010 ± 0.005 U/mg protein). CONCLUSION: The polysaccharide-rich extract of Caesalpinia ferrea stem barks present anti-inflammatory and antioxidant effects in mice models of acute inflammation induced by zymosan.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Antioxidants/pharmacology , Caesalpinia , Edema/prevention & control , Inflammation Mediators/metabolism , Oxidative Stress/drug effects , Peritonitis/prevention & control , Plant Bark , Plant Stems , Polysaccharides/pharmacology , Analgesics/pharmacology , Animals , Anti-Inflammatory Agents/isolation & purification , Antioxidants/isolation & purification , Biomarkers/metabolism , Caesalpinia/chemistry , Disease Models, Animal , Edema/chemically induced , Edema/metabolism , Edema/pathology , Female , Mice , Neutrophil Infiltration , Nociceptive Pain/chemically induced , Nociceptive Pain/metabolism , Nociceptive Pain/prevention & control , Peritonitis/chemically induced , Peritonitis/metabolism , Peritonitis/pathology , Plant Bark/chemistry , Plant Stems/chemistry , Polysaccharides/isolation & purification , Signal Transduction , ZymosanABSTRACT
Rheumatoid arthritis is an autoimmune inflammatory disease with progressive degradation of cartilage and joints. Additionally, gastric ulcer affects many patients who make prolonged use of non-steroidal anti-inflammatory drugs widely used in the symptomatic treatment of rheumatoid arthritis. Nerolidol, a natural sesquiterpene, has several biological activities including anti-inflammatory and antiulcerogenic action. This study aims to develop and characterize a nerolidol ß-cyclodextrin inclusion complex and to evaluate its activity in an experimental arthritis model. Inclusion complex was prepared by the lyophilization method and characterized by NMR, term analysis, XRD and SEM. Neutrophil migration assays and histopathological analysis were performed on zymosan-induced arthritis model using Swiss mice. And the gastroprotective effect was evaluated in two models of gastric ulcers: induced by ethanol and indomethacin. Inclusion complex showed no cytotoxicity and free nerolidol at a dose of 100 mg/kg (p.o.) in the arthritis model reduced neutrophil migration in 56% in relation to vehicle, and this inhibition was more expressive in the inclusion complex (67%) at the same dose. Histopathological analysis of the joint tissue confirmed the reduction of inflammatory signs. In the ethanol-induced gastric ulcer model, free nerolidol reduced the relative ulcer area more expressively (4.64%) than the inclusion complex (21.3%). However, in the indomethacin induction model, the inclusion complex showed better results in gastric protection compared to free nerolidol. The action of nerolidol complexed in beta-cyclodextrin in reducing arthritis inflammation combined with its gastroprotective action make it a potential new drug.
Subject(s)
Arthritis/drug therapy , Sesquiterpenes/pharmacology , beta-Cyclodextrins/pharmacology , Animals , Anti-Inflammatory Agents/pharmacology , Anti-Ulcer Agents/pharmacology , Arthritis, Experimental/drug therapy , Cell Line , Gastric Mucosa/metabolism , Indomethacin/pharmacology , Male , Mice , Plant Extracts/pharmacology , Stomach/pathology , Stomach Ulcer/chemically inducedABSTRACT
BACKGROUND: Naringenin is a biologically active analgesic, anti-inflammatory, and antioxidant flavonoid. Naringenin targets in inflammation-induced articular pain remain poorly explored. METHODS: The present study investigated the cellular and molecular mechanisms involved in the analgesic/anti-inflammatory effects of naringenin in zymosan-induced arthritis. Mice were pre-treated orally with naringenin (16.7-150 mg/kg), followed by intra-articular injection of zymosan. Articular mechanical hyperalgesia and oedema, leucocyte recruitment to synovial cavity, histopathology, expression/production of pro- and anti-inflammatory mediators and NFκB activation, inflammasome component expression, and oxidative stress were evaluated. RESULTS: Naringenin inhibited articular pain and oedema in a dose-dependent manner. The dose of 50 mg/kg inhibited leucocyte recruitment, histopathological alterations, NFκB activation, and NFκB-dependent pro-inflammatory cytokines (TNF-α, IL-1ß, and IL-33), and preproET-1 mRNA expression, but increased anti-inflammatory IL-10. Naringenin also inhibited inflammasome upregulation (reduced Nlrp3, ASC, caspase-1, and pro-IL-1ß mRNA expression) and oxidative stress (reduced gp91phox mRNA expression and superoxide anion production, increased GSH levels, induced Nrf2 protein in CD45+ hematopoietic recruited cells, and induced Nrf2 and HO-1 mRNA expression). CONCLUSIONS: Naringenin presents analgesic and anti-inflammatory effects in zymosan-induced arthritis by targeting its main physiopathological mechanisms. These data highlight this flavonoid as an interesting therapeutic compound to treat joint inflammation, deserving additional pre-clinical and clinical studies.
Subject(s)
Arthritis/drug therapy , Flavanones/therapeutic use , Leukocyte Common Antigens/analysis , NF-E2-Related Factor 2/physiology , Zymosan/pharmacology , Animals , Cytokines/biosynthesis , Disease Models, Animal , Dose-Response Relationship, Drug , Flavanones/pharmacology , Hematopoietic Stem Cells/metabolism , Inflammasomes/drug effects , Knee Joint/pathology , Male , Mice , NF-E2-Related Factor 2/genetics , NF-kappa B/physiology , Oxidative Stress/drug effects , Signal TransductionABSTRACT
Development of active compounds to control inflammation against systemic inflammatory response syndrome (SIRS) is critical important. Dioscin shows anti-inflammatory effects in our previous works. However, the action of the compound on SIRS still remained unknown. In the present paper, zymosan induced generalized inflammation (ZIGI) models in mice and rats, and PMA-differentiated THP1 cells stimulated by lipopolysaccharide (LPS) and Pam3-Cys-Ser-Lys4 (Pam3CSK4) were used. The results showed that dioscin significantly inhibited the proliferation of THP1 cells stimulated by LPS and Pam3CSK4, obviously reduced the soakage of inflammatory cells and necrosis in liver, kidney and intestine of rats and mice, and reduced peritoneal ascites fluid compared with ZIGI model groups. In addition, dioscin significantly declined the levels of alanine transaminase (ALT), aspartate transaminase (AST), creatinine (Cr), blood urea nitrogen (BUN), malondialdehyde (MDA) and myeloperoxidase (MPO), increased the levels of superoxide dismutase (SOD) in rats and mice. The migration of macrophages in tissues was also suppressed by dioscin. Mechanism investigation showed that dioscin significantly inhibited the expression levels of TLR2, MyD88, NFκb, HMGB1, increased the expression levels of IKBα, and decreased the mRNA levels of interleukin1 beta (IL1ß), interleukin6 (IL6) and tumor necrosis factoralpha (TNFα) in liver, kidney, intestine tissues of rats and mice, and in PMA-differentiated THP1 cells, which were further confirmed by TLR2 siRNA silencing in vitro. In conclusion, our data confirmed that dioscin exhibited protective effects against SIRS via adjusting TLR2/MyD88 signal pathway, which should be developed as one potent candidate to treat SIRS in the future.
Subject(s)
Anti-Inflammatory Agents/therapeutic use , Diosgenin/analogs & derivatives , Macrophages/drug effects , Animals , Dioscorea/immunology , Diosgenin/therapeutic use , Humans , Macrophages/physiology , Male , Mice , Mice, Inbred C57BL , Myeloid Differentiation Factor 88/metabolism , NF-kappa B/metabolism , Rats , Rats, Sprague-Dawley , Signal Transduction/drug effects , Systemic Inflammatory Response Syndrome/drug therapy , THP-1 Cells , Toll-Like Receptor 2/metabolism , Zymosan/immunologyABSTRACT
Irritable bowel syndrome (IBS) is a common gastrointestinal disorder manifesting as unexplained abdominal pain and bowel habit changes. The pathogenesis of post-infectious IBS is associated with gutâ»brain axis dysfunction, including low-grade colonic inflammation and anxiety-related long-term brain changes. This study analyzed the efficacy of a standardized extract of Dolichos lablab L. extract (DL), a bean species, in an IBS mouse model resembling post-infectious, diarrhea-dominant IBS. Using a zymosan-induced animal IBS model, we found that oral administration of DL significantly attenuated zymosan-induced increases in colonic macroscopic scores and minimized weight loss without affecting food intake. In the DL-treated mice, the mast cell count and tumor necrosis factor-α level in the colon markedly decreased, similar to results in sulfasalazine-treated mice and in mice with lipopolysaccharide-stimulated bone marrow-derived mast cells. The number of visceral pain-related behaviors was much lower in the DL-treated mice. Anxiety-like behaviors significantly improved, comparable to that after treatment with amitriptyline. The c-Fos expression level in the prefrontal cortex was significantly reduced. Our data suggest that DL could be beneficial for treating IBS by acting on the gut and brain.
Subject(s)
Dolichos/chemistry , Irritable Bowel Syndrome/drug therapy , Phytotherapy/methods , Plant Extracts/administration & dosage , Administration, Oral , Animals , Anxiety/drug therapy , Anxiety/etiology , Brain/metabolism , Colon/metabolism , Disease Models, Animal , Irritable Bowel Syndrome/chemically induced , Irritable Bowel Syndrome/psychology , Male , Mice , Mice, Inbred C57BL , ZymosanABSTRACT
Spleen tyrosine kinase (Syk), a non-receptor tyrosine kinase, plays an important role in allergic diseases and inflammation. Syk triggers several intracellular signalling cascades including Toll-like receptor signalling to activate inflammatory responses following fungal infection but the role of this enzyme in zymosan (ZYM)-induced non-septic shock and its impacts on hypotension and inflammation in rats is not well understood. This study was conducted to determine the effects of Syk inhibition on ZYM-induced alterations in the expression and/or activities of Syk, inhibitor ĸB (IĸB)-α, and nuclear factor-ĸB (NF-ĸB) p65. We also examined the effect of Syk inhibition on inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2, and tumour necrosis factor (TNF)-α, and activity of myeloperoxidase (MPO) that contribute to hypotension and inflammation. Administration of ZYM (500 mg/kg, ip) to male Wistar rats decreased blood pressure and increased heart rate. These changes were associated with increased expression and/or activities of Syk, NF-κB p65, iNOS and COX-2 and decreased expression of IκB-α with enhanced levels of nitrite, nitrotyrosine, 6-keto-PGF1α , and TNF-α and activity of MPO in renal, cardiac and vascular tissues. ZYM administration also elevated serum and tissue nitrite levels. The selective Syk inhibitor BAY 61-3606 (3 mg/kg, ip) given 1 hour after ZYM injection reversed all of these changes induced by ZYM. These results suggest that Syk/IĸB-α/NF-ĸB pathway activation contributes to hypotension and inflammation caused by the production of vasodilator and proinflammatory mediators in the zymosan-induced non-septic shock model.
Subject(s)
I-kappa B Kinase/metabolism , NF-kappa B/metabolism , Niacinamide/analogs & derivatives , Pyrimidines/therapeutic use , Shock/chemically induced , Syk Kinase/metabolism , Animals , Gene Expression Regulation/drug effects , I-kappa B Kinase/genetics , Male , NF-kappa B/genetics , Niacinamide/therapeutic use , Rats , Rats, Wistar , Shock/drug therapy , Syk Kinase/antagonists & inhibitors , Syk Kinase/genetics , Zymosan/toxicityABSTRACT
Irritable bowel syndrome (IBS) is a functional gastrointestinal disease with complex pathophysiology involving the brain-gut axis. To assess the effects of Wasabia koreana (WK) on IBS, we employed a mouse model of colonic zymosan injection presenting with diarrhea-predominant IBS-like symptoms. Oral WK administration significantly diminished stool score, suppressed colon length and weight change, and minimized body weight loss without affecting food intake. In WK-treated mice, the submucosal thickening and epithelial lining of the colon were inhibited and were similar to those of naïve mice. Infiltration of mast cells into the colon and serum tumor necrosis factor-α levels were markedly suppressed. These effects were comparable to those of sulfasalazine, an anti-inflammatory drug. Furthermore, the number of visceral pain-related behaviors was significantly decreased, and locomotion activities measured in the elevated plus maze and open field tests were significantly increased by WK in a dose-dependent manner compared with amitriptyline, an antidepressant. These changes were accompanied by reduced FosB2 expression in the brain. Taken together, these data suggest that WK may have potential as a medicinal food for IBS by acting on inflammatory diarrhea and neural activity.
Subject(s)
Irritable Bowel Syndrome/drug therapy , Plant Extracts/administration & dosage , Wasabia/chemistry , Zymosan/adverse effects , Animals , Colon/drug effects , Colon/immunology , Disease Models, Animal , Humans , Irritable Bowel Syndrome/chemically induced , Irritable Bowel Syndrome/immunology , Male , Mice , Mice, Inbred C57BL , Plant Extracts/analysis , Tumor Necrosis Factor-alpha/immunologyABSTRACT
Oxidative stress is harmful to the microbes but also to the host, and may result in bystander damage or death. Because of this, respiratory burst triggered in phagocytes by pathogens is counteracted by production of antioxidative factors. The aim of this work was to examine effectiveness of the latter system in earthworms Eisenia andrei by induction of reactive oxygen species, lipofuscin and phenoloxidase by natural (LPS, zymosan, Micrococus luteus) and synthetic (phorbol ester, PMA) stimulants. The compounds impaired numbers, viability (increased apoptosis) and composition of coelomocytes, and triggered the antioxidant activity of catalase and selenium-dependent glutathione peroxidase. The natural pathogenic compounds, unlike PMA, strongly activated antioxidative responses that diminished cell apoptosis. Moreover, repeated exposure to the same or different pathogenic compounds did not induce respiratory burst exhausted phenotype showing that coelomocytes are constantly at bay to withstand numerous infections. The current study reveals importance and efficiency of the oxidative-antioxidative systems in annelids but also confirms its evolutionary conservatism and complexity even in lower taxa of the animal kingdom.
Subject(s)
Antioxidants/metabolism , Apoptosis/immunology , Oligochaeta/cytology , Oligochaeta/physiology , Adjuvants, Immunologic/pharmacology , Animals , Apoptosis/drug effects , Catalase/metabolism , Cell Survival/drug effects , Cell Survival/immunology , Glutathione Peroxidase/metabolism , Lipofuscin/metabolism , Membrane Potential, Mitochondrial/drug effects , Membrane Potential, Mitochondrial/immunology , Monophenol Monooxygenase/metabolism , Oligochaeta/drug effects , Reactive Oxygen Species/metabolism , Respiratory Burst/drug effects , Respiratory Burst/immunologyABSTRACT
Although sigma-1 receptor (Sig-1R) antagonists have a potential antinociceptive effect in inflammatory diseases, the precise mechanism is not fully understood. The present study was aimed to elucidate the role of spinal neurons and microglia in the anti-nociceptive mechanism of BD1047 (a prototypical Sig-1R antagonist) using an inflammatory pain model based on intraplantar injection of zymosan. Oral pretreatment with BD1047 dose-dependently reduced zymosan-induced thermal and mechanical hyperalgesia as well as spinal neuronal activation including increased immunoreactivity of Fos, protein kinase C (PKC) and 'PKC-dependent phosphorylation of the NMDA receptor subunit 1' (pNR1). Zymosan also led to increased CD11b immunoreactivity (a marker of microglia) accompanied by 'phosphorylated p38 mitogen activated protein kinase' (p-p38MAPK) and interleukin-1ßimmunoreactivity in the spinal dorsal horn. Intrathecal injection of a microglia modulator (minocycline), p38MAPK inhibitor (SB203580) or interleukin-1ßneutralizing antibody significantly attenuated zymosan-induced hyperalgesia. Specifically, oral pretreatment with BD1047 reduced the immunoreactivity of CD11b, p-p38MAPK and interleukin-1ß. In the spinal cord section, Sig-1R immunoreactivity was exclusively distributed in both spinal dorsal horn neurons and central endings of unmyelinated primary afferent fibers but not in glia. Intrathecal injection of BD1047 alleviated zymosan-induced hyperalgesia up to the level of oral administration. Taken together, our data imply that antinociceptive effect induced by oral treatment with BD1047 may be mediated, at least in part, by the inhibition of neuronal and microglial activation in the spinal cord triggered by inflammatory conditions.
Subject(s)
Analgesics/pharmacology , Ethylenediamines/pharmacology , Pain/drug therapy , Pain/metabolism , Spinal Cord/drug effects , Spinal Cord/metabolism , Administration, Oral , Animals , Anti-Inflammatory Agents/pharmacology , Disease Models, Animal , Dose-Response Relationship, Drug , Drug Evaluation, Preclinical , Hyperalgesia/drug therapy , Hyperalgesia/metabolism , Hyperalgesia/pathology , Male , Microglia/drug effects , Microglia/metabolism , Microglia/pathology , Pain/pathology , Posterior Horn Cells/drug effects , Posterior Horn Cells/metabolism , Posterior Horn Cells/pathology , Rats, Sprague-Dawley , Receptors, sigma/antagonists & inhibitors , Receptors, sigma/metabolism , Spinal Cord/pathology , ZymosanABSTRACT
The ethyl acetate extract from the fruit pulp of Caryocar coriaceum Wittm (Caryocaraceae), popularly known as pequi, has wide applications in popular medicine. Preclinical tests have demonstrated the therapeutic properties of the oil. We investigated the antinociceptive and anti-inflammatory effects of Pequi C. coriaceum Wittm ethyl acetate extract (PCCO) on zymosan-induced arthritis in rat knee joint. The animals were pretreated with PCCO for 7 consecutive days or with a single dose. Paw elevation time (PET), leukocyte infiltration, myeloperoxidase activity (MPO) and cytokine levels were assessed 4h after zymosan injection. Synovial tissue was harvested for immunohistochemical analysis, edema and vascular permeability. We observed a significant decrease in PET with PCCO pretreatment. PCCO showed a significant reduction of leukocyte migration and a decrease in MPO. Decreases were observed in cytokine release in the synovial fluid and TNF-α and cyclooxygenase-1 immunostaining in synovial tissue. Edema was inhibited by treatment with all doses of PCCO. The data suggest that PCCO exerts antinociceptive and anti-inflammatory effects on arthritis in rats.
Subject(s)
Analgesics/pharmacology , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Arthritis, Experimental/drug therapy , Ericales/chemistry , Fruit/chemistry , Plant Extracts/chemistry , Plant Extracts/pharmacology , Analgesics/isolation & purification , Animals , Anti-Inflammatory Agents, Non-Steroidal/isolation & purification , Arthritis, Experimental/chemically induced , Arthritis, Experimental/pathology , Cyclooxygenase 1/metabolism , Cytokines/metabolism , Edema/chemically induced , Edema/drug therapy , Edema/pathology , Joints/pathology , Male , Neutrophil Infiltration/drug effects , Peroxidase/metabolism , Rats , Rats, Wistar , Synovial Fluid/drug effects , Synovial Fluid/metabolism , Tumor Necrosis Factor-alpha/metabolism , ZymosanABSTRACT
In the present study, we assessed whether 7-hydroxycoumarin (umbelliferone), 7-hydroxy-4-methylcoumarin, and their acetylated analogs modulate some of the effector functions of human neutrophils and display antioxidant activity. These compounds decreased the ability of neutrophils to generate superoxide anion, release primary granule enzymes, and kill Candida albicans. Cytotoxicity did not mediate their inhibitory effect, at least under the assessed conditions. These coumarins scavenged hypochlorous acid and protected ascorbic acid from electrochemical oxidation in cell-free systems. On the other hand, the four coumarins increased the luminol-enhanced chemiluminescence of human neutrophils stimulated with phorbol-12-myristate-13-acetate and serum-opsonized zymosan. Oxidation of the hydroxylated coumarins by the neutrophil myeloperoxidase produced highly reactive coumarin radical intermediates, which mediated the prooxidant effect observed in the luminol-enhanced chemiluminescence assay. These species also oxidized ascorbic acid and the spin traps α-(4-pyridyl-1-oxide)-N-tert-butylnitrone and 5-dimethyl-1-pyrroline-N-oxide. Therefore, 7-hydroxycoumarin and the derivatives investigated here were able to modulate the effector functions of human neutrophils and scavenge reactive oxidizing species; they also generated reactive coumarin derivatives in the presence of myeloperoxidase. Acetylation of the free hydroxyl group, but not addition of the 4-methyl group, suppressed the biological effects of 7-hydroxycoumarin. These findings help clarify how 7-hydroxycoumarin acts on neutrophils to produce relevant anti-inflammatory effects.
Subject(s)
Antifungal Agents/pharmacology , Antioxidants/pharmacology , Candida albicans/drug effects , Neutrophils/drug effects , Umbelliferones/pharmacology , Anions/antagonists & inhibitors , Anions/metabolism , Antifungal Agents/chemistry , Antioxidants/chemistry , Dose-Response Relationship, Drug , Humans , Microbial Sensitivity Tests , Molecular Structure , Neutrophils/metabolism , Reactive Oxygen Species/antagonists & inhibitors , Reactive Oxygen Species/metabolism , Structure-Activity Relationship , Umbelliferones/chemistryABSTRACT
Many cells in the organism besides pinealocytes, synthesize melatonin. Here, we evaluate both the mechanism of zymosan-induced melatonin synthesis and its autocrine effect in human colostral mononuclear cells. The synthesis of melatonin was induced by activation of the transcription factor nuclear factor kappa B (NF-κB), as either the blockade of the proteasome or the binding of NF-κB to DNA inhibits zymosan-induced melatonin synthesis. As observed in RAW 264.7 lineage cells, the dimer involved is RelA/c-Rel. Melatonin plays a direct role in mononuclear cell activity, increasing zymosan-induced phagocytosis by stimulating MT2 melatonin receptors and increasing the expression of dectin-1. This role was confirmed by the blockade of melatonin receptors using the competitive antagonist luzindole and the MT2 -selective partial agonist 4P-PDOT. In summary, we show that melatonin produced by immune-competent cells acts in an autocrine manner, enhancing the clearance of pathogens by increasing phagocyte efficiency. Given that these cells are present in human colostrum for 4 or 5 days after birth, this mechanism may be relevant for the protection of infant health.