RESUMO
Orofacial pain has significant psychological and physiological effects. Citral (3,7-dimethyl-2,6-octadienal) is the main component of Cymbopogon citratus (DC) Stapf, an herb with analgesic properties. Although citral has been considered a potent analgesic, its putative effects on orofacial pain are still unknown. OBJECTIVE: The objective of this study is to test the hypothesis that citral modulates orofacial pain using two experimental models: formalin-induced hyperalgesia in the vibrissae area and during persistent temporomandibular hypernociception using Complete Freund's Adjuvant - CFA test. METHODS: For the formalin test, citral (100 and 300 mg/kg, oral gavage) or its vehicle (Tween 80, 1 %) were given 1 h before the formalin injection subcutaneously (sc) into the vibrissae area. For the CFA model, we analyzed the prophylactic (100 mg/kg of citral by oral gavage, 1 h before CFA injection) and the chronic therapeutic (citral treatment 1-hour post-CFA injection and daily post-CFA injection) effect of citral or its vehicle in animals treated with CFA for 8 days. RESULTS: Citral caused a decrease in formalin-induced local inflammation and the time spent performing nociceptive behavior in a dose-dependent fashion. Similarly, prophylactic and therapeutic citral treatment decreased the CFA-induced persistent mechanical hypernociception in the temporomandibular area. CONCLUSION: Our data strengthen the notion that citral plays a powerful antinociceptive role by decreasing orofacial hypernociception in formalin and CFA models.
Assuntos
Dor Facial , Hiperalgesia , Ratos , Animais , Hiperalgesia/tratamento farmacológico , Dor Facial/tratamento farmacológico , Dor Facial/etiologia , Inflamação/tratamento farmacológico , Inflamação/induzido quimicamente , Analgésicos/farmacologia , FormaldeídoRESUMO
Studies performed in a mouse model of chronic inflammatory pain induced by intraplantar injection of complete Freund's adjuvant (CFA) have shown that constitutive activation of the endogenous opioid signaling, besides serving as a mechanism of endogenous analgesia that tonically represses pain sensitization, also generates a state of endogenous opioid dependence. Since species-related differences concerning pain biology and addictive behaviors occur between mice and rats, the present study explored whether the coexistence of endogenous opioid analgesia and endogenous opioid dependence also characterizes a homologous rat model. To this aim, CFA-injured Wistar rats were treated with either 3 mg/kg or 10 mg/kg of the opioid receptor inverse agonist naltrexone (NTX) during the pain remission phase and monitored for 60 min for possible withdrawal behaviors. At 3 mg/kg, NTX, besides inducing the reinstatement of mechanical allodynia, also caused a distinct appearance of ptosis, with slight but nonsignificant changes to the occurrence of teeth chatters and rearing. On the other hand, 10 mg/kg of NTX failed to unmask pain sensitization and induced significantly lower levels of ptosis than 3 mg/kg. Such an NTX-related response pattern observed in the rat CFA model seems to differ substantially from the pattern previously described in the mouse CFA model. This supports the knowledge that mice and rats are not identical in terms of pharmacological response and stresses the importance of choosing the appropriate species for preclinical pain research purposes depending on the scientific question being asked.
Assuntos
Dor Crônica , Transtornos Relacionados ao Uso de Opioides , Ratos , Camundongos , Animais , Analgésicos Opioides/farmacologia , Agonismo Inverso de Drogas , Ratos Wistar , Inflamação/tratamento farmacológico , Dor Crônica/tratamento farmacológico , Hiperalgesia/tratamento farmacológico , Peptídeos Opioides/uso terapêutico , Naltrexona/farmacologia , Naltrexona/uso terapêutico , Transtornos Relacionados ao Uso de Opioides/tratamento farmacológico , Modelos Animais de DoençasRESUMO
OBJECTIVE: This study aimed to evaluate whether ligature-induced periodontitis and rheumatoid arthritis (RA) potentiate the deleterious effects on functional capacity, periodontal and synovial tissues, leukocyte migration, and interleukin 17 (IL-17) levels, and to investigate the repercussions of single Freund's Complete Adjuvant (FCA) injection associated with periodontitis. MATERIALS AND METHODS: Fifty-one male Wistar rats were randomised into six groups: control (CG, n = 8), RA (RAG, n = 9), periodontitis (PG, n = 9), periodontitis and RA (PRAG, n = 9), periodontitis and intradermal injection (PIDG, n = 9), and periodontitis and intra-articular injection (PIAG, n = 7). The animals underwent ligature placement and one or two injections with FCA to induce RA. Motor disability, nociceptive threshold, joint edema, and muscle strength were assessed, and the animals were euthanized on day 30. Synovial fluid, hemimandibles, and knee joints were collected. RESULTS: PRAG showed no reduction of edema or improvement of muscle strength, whereas it showed most significant changes in leukocyte migration, morphological analyses of the synovial membrane (SM), and radiographic and histometric analyses of the jaw. The PIAG showed some alterations, though not permanent. CONCLUSION: Ligature-induced periodontitis and RA induced by two FCA injections accentuated the deleterious effects on functional capacity, leukocyte migration, synovial and periodontal tissues.
Assuntos
Artrite Reumatoide , Periodontite , Animais , Masculino , Ratos , Artrite Reumatoide/complicações , Edema/induzido quimicamente , Leucócitos , Modelos Teóricos , Periodontite/complicações , Ratos Wistar , Movimento Celular , Interleucina-17RESUMO
Inflammatory pain associates with spinal glial activation and central sensitization. Systemic administration of IMT504, a non-CpG oligodeoxynucleotide originally designed as an immunomodulator, exerts remarkable anti-allodynic effects in rats with complete Freund´s adjuvant (CFA)-induced hindpaw inflammation. However, the anti-nociceptive mechanisms of IMT504 remain unknown. Here we evaluated whether IMT504 blocks inflammatory pain-like behavior by modulation of spinal glia and central sensitization. The study was performed in Sprague Dawley rats with intraplantar CFA, and a single lumbosacral intrathecal (i.t.) administration of IMT504 or vehicle was chosen to address if changes in glial activation and spinal sensitization relate to the pain-like behavior reducing effects of the ODN. Naïve rats were also included. Von Frey and Randall-Selitto tests, respectively, exposed significant reductions in allodynia and mechanical hypersensitivity, lasting at least 24 h after i.t. IMT504. Analysis of electromyographic responses to electrical stimulation of C fibers showed progressive reductions in wind-up responses. Accordingly, IMT504 significantly downregulated spinal glial activation, as shown by reductions in the protein expression of glial fibrillary acidic protein, CD11b/c, Toll-like receptor 4 (TLR4) and the phosphorylated p65 subunit of NFκB, evaluated by immunohistochemistry and western blot. In vitro experiments using early post-natal cortical glial cultures provided further support to in vivo data and demonstrated IMT504 internalization into microglia and astrocytes. Altogether, our study provides new evidence on the central mechanisms of anti-nociception by IMT504 upon intrathecal application, and further supports its value as a novel anti-inflammatory ODN with actions upon glial cells and the TLR4/NFκB pathway. Intrathecal administration of the non-CpG ODN IMT504 fully blocks CFA-induced mechanical allodynia and hypersensitivity, in association with reduced spinal sensitization. Administration of the ODN also results in downregulated gliosis and reduced TLR4-NF-κB pathway activation. IMT504 uptake into astrocytes and microglia support the concept of direct modulation of CFA-induced glial activation.
Assuntos
Sensibilização do Sistema Nervoso Central , Hiperalgesia , Animais , Hiperalgesia/tratamento farmacológico , Inflamação , Oligodesoxirribonucleotídeos , Dor , Ratos , Ratos Sprague-Dawley , Medula EspinalRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: An infusion obtained from the leaves of "chal-chal" (Allophylus edulis Radlk.) is used for popular treatment of intestinal disorders and as an anti-inflammatory throat treatment. Because of the anti-inflammatory medicinal folk use, a previous work reported scientific research confirming the anti-inflammatory activity of A. edulis essential oil collected in Dourados, MS, Brazil, in March 2015. AIM OF THE STUDY: The aim of this study was to evaluate the variation in the chemical profile of the essential oil of A. edulis plants collected in Dourados (EOAE-D) and Bonito (EOAE-B), two cities in Mato Grosso do Sul State, Brazil. Additionally, we evaluated the anti-inflammatory effects of the essential oil, as well as that of the major compounds (caryophyllene oxide and α-zingiberene), in experimental in vivo models of inflammation in mice. MATERIALS AND METHODS: Leaves were collected from plants at both sites in July 2018. The composition of the essential oil (EOAE-D and EOAE-B) was determined by GC/MS, and major compounds (caryophyllene oxide and α-zingiberene) were isolated and identified by chromatographic methods and NMR spectroscopy. Anti-inflammatory capacities were assessed using two classical models of inflammatory models, carrageenan- and CFA-induced paw inflammation (mechanical and thermal hyperalgesia). RESULTS: Both EOAE-D and EOAE-B showed sesquiterpenes as a major constituent, namely, caryophyllene oxide (29.5%) and α-zingiberene (45.0%), respectively. In tests, EOAE, caryophyllene oxide and α-zingiberene-induced antiedematogenic and antihyperalgesic effects were found in the different utilized models. CONCLUSIONS: The results indicate that samples from the two cities differed in chemical composition but not in their anti-inflammatory and antihyperalgesic effects. This finding corroborates the use of A. edulis as a medicinal plant and indicates its potential in the therapy of inflammatory conditions.
Assuntos
Anti-Inflamatórios/farmacologia , Hiperalgesia/prevenção & controle , Inflamação/prevenção & controle , Óleos Voláteis/farmacologia , Folhas de Planta , Óleos de Plantas/farmacologia , Sapindaceae , Animais , Anti-Inflamatórios/isolamento & purificação , Brasil , Carragenina , Modelos Animais de Doenças , Hiperalgesia/induzido quimicamente , Hiperalgesia/microbiologia , Hiperalgesia/fisiopatologia , Inflamação/induzido quimicamente , Inflamação/microbiologia , Inflamação/fisiopatologia , Masculino , Camundongos Endogâmicos C57BL , Mycobacterium tuberculosis , Óleos Voláteis/isolamento & purificação , Limiar da Dor/efeitos dos fármacos , Folhas de Planta/química , Óleos de Plantas/isolamento & purificação , Sesquiterpenos Policíclicos/farmacologia , Polifenóis/farmacologia , Sapindaceae/químicaRESUMO
IMT504 is a non-CPG, non-coding synthetic oligodeoxinucleotide (ODN) with immunomodulatory properties and a novel inhibitory role in pain transmission, exerting long-lasting analgesic effects upon multiple systemic administrations. However, its mechanisms of anti-nociceptive action are still poorly understood. In the present study in male adult rats undergoing complete Freund's adjuvant-induced hindpaw inflammation, we focused in the analysis of the immunomodulatory role of IMT504 over the cellular infiltrate, the impact on the inflammatory milieu, and the correlation with its anti-allodynic role. By means of behavioral analysis, we determined that a single subcutaneous administration of 6 mg/kg of IMT504 is sufficient to exert a 6-week-long full reversal of mechanical and cold allodynia, compromising neither acute pain perception nor locomotor activity. Importantly, we found that the anti-nociceptive effects of systemic IMT504, plus quick reductions in hindpaw edema, were associated with a modulatory action upon cellular infiltrate of B-cells, macrophages and CD8+ T-cells populations. Accordingly, we observed a profound downregulation of several inflammatory leukocyte adhesion proteins, chemokines and cytokines, as well as of ß-endorphin and an increase in the anti-inflammatory cytokine, interleukin-10. Altogether, we demonstrate that at least part of the anti-nociceptive actions of IMT504 relate to the modulation of the peripheral immune system at the site of injury, favoring a switch from pro- to anti-inflammatory conditions, and provide further support to its use against chronic inflammatory pain. Graphical abstract GA short description - IMT504 systemic Administration. Systemic administration of the non-CpG ODN IMT504 results in a 6-week long blockade of pain-like behavior in association with anti-inflammatory responses at the site of injury. These include modulation of lymphoid and myeloid populations plus downregulated expression levels of multiple pro-inflammatory cytokines and ß-endorphin. Nocifensive responses and locomotion remain unaltered.
Assuntos
Analgesia , Dor Crônica , Animais , Linfócitos T CD8-Positivos , Dor Crônica/tratamento farmacológico , Modelos Animais de Doenças , Hiperalgesia , Inflamação/tratamento farmacológico , Masculino , Oligodesoxirribonucleotídeos , RatosRESUMO
Abstract Objectives To investigate the effect of a standardized extract of Centella asiatica (ECa 233), which has anti-inflammatory properties, on the local expression of the transient receptor potential vanilloid 1 (TRPV1), the acid-sensing ion channel subunit 3 (ASIC3), and the calcitonin gene-related peptide (CGRP) in the temporomandibular joint (TMJ) structure 21 days after injecting the TMJ with complete Freund's adjuvant (CFA). Methodology A mouse model was induced by analyzing the CFA-injected TMJ on days 7, 14, and 21. We assessed TMJ histology by the osteoarthritis cartilage grade score. Then, we observed the effect of different ECa 233 concentrations (30, 100, and 300 mg/kg) and of 140 mg/kg ibuprofen doses on TRPV1, ASIC3, and CGRP local expression on day 21. Results Osteoarthritis cartilage scores were 1.17±0.37 and 3.83±0.68 on days 14 and 21, respectively, in the CFA group (n=5). On day 21, TRPV1, ASIC3, and CGRP expression significantly increased in the CFA group. In the ibuprofen-treated group, TRPV1 expression significantly decreased, but ASIC3 and CGRP showed no significant difference. All ECa 233 doses reduced TRPV1 expression, but the 100 mg/kg ECa 233 dose significantly decreased ASIC3 expression. Conclusions TRPV1, ASIC3, and CGRP expression increased in mice with TMJ-OA on day 21. All ECa 233 and ibuprofen doses inhibited pathogenesis by modulating the local expression of TRPV1 and ASIC3. Therefore, ECa 233 was more effective than ibuprofen.
Assuntos
Animais , Coelhos , Osteoartrite/tratamento farmacológico , Centella , Articulação Temporomandibular , Extratos Vegetais/farmacologia , Mediadores da InflamaçãoRESUMO
Abstract Mirror-image pain is a kind of pain that occurs on the contralateral side, but its pathogenesis remains unclear. Objective To develop an osteoarthritis mouse model for investigating mirror-image pain through observing nocifensive behaviors, histological changes, and nociceptive activity at days 3, 7, 14, 21, and 28 after the chemical induction of unilateral temporomandibular joint (TMJ) osteoarthritis. Methodology We randomly divided 6-week-old mice into sham and complete Freund adjuvant groups. To induce nocifensive behaviors, we applied 0.04 g of von Frey filament, 10 psi of air puff, and cold acetone on both sides of whisker pads at different days. The histology of TMJ on both sides was observed by hematoxylin/eosin staining and microcomputed tomography scanning. Furthermore, the nociceptive activity was evaluated using the phosphorylated cyclic AMP response element binding protein (pCREB) and a microglia marker at different days in the trigeminal subnucleus caudalis. Results Nocifensive behaviors against mechanical and temperature stimuli on the contralateral side became stronger than the baseline on day 28, in agreement with the elevation of the pCREB and the microglia marker in the trigeminal subnucleus caudalis. Thus, hypernociception on the contralateral side occurred at day 28. Conclusions Clearly, the TMJ model with unilateral osteoarthritis exhibited mirror-image pain. Therefore, this model is useful in investigating the pathogenesis of pain and in developing treatments.
Assuntos
Animais , Camundongos , Osteoartrite/diagnóstico por imagem , Articulação Temporomandibular , Dor , Adjuvante de Freund , Microtomografia por Raio-XRESUMO
AIM: The present work aimed at the DIM-loaded microparticles development and anti-hypernociceptive action evaluation. METHOD: The formulations were prepared by O/W solvent emulsion-evaporation method and characterised by particle diameter, content and DIM encapsulation efficiency, drug release profile, thermal behaviour and physicochemical state. The anti-hypernociceptive action was evaluated in the animal model of acute inflammatory pain. RESULT: The MPs had a mean diameter in the micrometric range (368 ± 31 µm), narrow size distribution, DIM content of 150 mg/g, encapsulation efficiency around 84% and prolonged compound release. Evaluations of the association form of DIM to MPs demonstrated the feasibility of the systems to incorporate DIM and increases its thermal stability. An improvement in the anti-hypernociceptive action of DIM was observed by its microencapsuation, because it was increased and prolonged. CONCLUSION: Therefore, the MPs developed represent a promising formulation for oral administration of the DIM in the treatment of inflammatory pain.
Assuntos
Anti-Inflamatórios não Esteroides/administração & dosagem , Celulose/análogos & derivados , Portadores de Fármacos/química , Indóis/administração & dosagem , Dor/tratamento farmacológico , Animais , Anti-Inflamatórios não Esteroides/uso terapêutico , Cápsulas , Celulose/química , Indóis/uso terapêutico , Masculino , CamundongosRESUMO
This study investigated the effects of systemic treatment with a new formulation of resveratrol (RSV) vehicled in rice oil (RSVO) in experimental rat models of inflammation. Male Wistar rats were evaluated in the following in vivo models: carrageenan-induced acute edema, complete Freund's adjuvant (CFA)-evoked sub-chronic edema, and CFA-induced polyarthritis. The animals were treated orally with RSVO (10-15 mg/kg) or RSV (100-200 mg/kg), depending on the experimental protocol. RSV was more effective than RSVO in carrageenan-elicited acute edema when dosed in either prophylactic or therapeutic schemes of administration. However, the repeated RSVO administration, at 10-fold lower doses, exhibited superior anti-inflammatory actions in either the sub-chronic edema or the chronic polyarthritis model elicited by CFA, when compared with RSV. The novel formulation RSVO displayed a lower plasma biotransformation when compared with the RSV-treated group-46% versus 88% of metabolites, respectively. RSVO also prevented polyarthritis-related cartilage destruction, an effect that might rely on the inhibition of the pro-inflammatory cytokine interleukin-6 (IL-6), associated with an increase of the anti-inflammatory cytokine interleukin-10 (IL-10). Noteworthy, the long-term administration of RSVO did not elicit any gastrointestinal harm. Our study revealed that RSVO was notably effective in the long-term inflammatory and degenerative responses triggered by CFA. This innovative formulation might well represent a promising alternative for treating chronic inflammatory diseases, such as arthritis.
Assuntos
Anti-Inflamatórios/farmacologia , Inflamação/prevenção & controle , Articulações/efeitos dos fármacos , Resveratrol/farmacologia , Óleo de Farelo de Arroz/farmacologia , Animais , Carragenina , Citocinas/metabolismo , Modelos Animais de Doenças , Composição de Medicamentos , Adjuvante de Freund , Inflamação/induzido quimicamente , Inflamação/metabolismo , Inflamação/patologia , Mediadores da Inflamação/metabolismo , Articulações/metabolismo , Articulações/patologia , Masculino , Ratos WistarRESUMO
This study aimed to analyze the antiarthritic activity of ginkgolic acid against the Complete Freund's Adjuvant (CFA)-induced arthritis in rats. Arthritis was induced through an intradermal injection of CFA (0.1 mL) at the right hind footpad of adult Wistar Albino rats. Ginkgolic acid was administered orally at doses of 25 mg/kg and 50 mg/kg, respectively, once daily via gavage for 25 days upon inducing arthritis. Indomethacin was administered orally at a dose of 3 mg/kg twice in a week which served as positive control group. The animals were sacrificed and subjected to biochemical and histopathological analysis upon completion of treatment. Ginkgolic acid was able to reverse the arthritic effect (p < 0.01) induced by CFA in a dose dependent manner. Swelling of paw, thymus and spleen index, serum biomarker levels, and pro-inflammatory cytokines were significantly reduced (p < 0.01) by the acid whereas the antioxidant enzyme activities were remarkably restored. The histopathological findings were in agreement with the biochemical results. The results indicate that the antioxidant and anti-inflammatory properties of ginkgolic acid can be credited to the antiarthritic effects, and it can be promoted as a potential agent for therapeutic use against osteoarthritis
Assuntos
Animais , Masculino , Ratos , Artrite Experimental/induzido quimicamente , Adjuvante de Freund/agonistas , Osteoartrite/patologia , Injeções Intradérmicas , Indometacina , Antioxidantes/classificaçãoRESUMO
The poor immunogenicity displayed by some antigens has encouraged the development of strategies to improve the immune response and safety of vaccine candidates, resulting in an intense search for substances that potentiate vaccine response. Adjuvants have these properties helping vaccine candidates to induce a strong, durable, and fast immune response. In this study, we evaluated the specific immune response of adjuvants alone, Saponin (SAP), Incomplete Freund's Adjuvant (IFA) and Monophosphoryl lipid-A SE (MPL-SE®) and in combination with total antigen of L. braziliensis (LB): LBSAP, LBIFA and LBMPL. The specific immune response induced by these compositions demonstrated that they were powerfully immunogenic, increasing cellular infiltration in the skin. Draining lymph nodes cultures showed that LBIFA and LBMPL have higher ability to increase the capacity of APCs to present antigens, with increased frequency of CD11c+CD86+ cells. SAP, MPL, LBSAP, LBIFA and LBMPL could activate lymphocytes increasing expression of CD69 and CD25. LBSAP group was an excellent inducer of pro-inflammatory cytokines at 24â¯h. At 48â¯h, higher cytokines production was observed in IFA, LBIFA, MPL and LBMPL groups. Our data demonstrate that LBSAP and LBMPL are potential formulations to be tested in other experimental models. Also, the data obtained could expand the knowledge about immune response after sensitization and also contribute to the development of safe, immunogenic and effective vaccines.
Assuntos
Adjuvantes Imunológicos/farmacologia , Antígenos de Protozoários/imunologia , Leishmania braziliensis/imunologia , Vacinas contra Leishmaniose/imunologia , Leishmaniose Visceral/prevenção & controle , Animais , Anticorpos Antiprotozoários/imunologia , Formação de Anticorpos/imunologia , Adjuvante de Freund/imunologia , Imunidade Inata/efeitos dos fármacos , Imunidade Inata/imunologia , Lipídeo A/análogos & derivados , Lipídeo A/imunologia , Lipídeos/imunologia , Masculino , Camundongos , Saponinas/imunologiaRESUMO
The effect of quercetin was assessed in rats induced with complete Freund adjuvant (CFA). Arthritis scores, paw oedema, latency, activities of myeloperoxidase (MPO), ectonucleoside triphosphate diphosphohydrolase (E-NTPDase), and ectoadenosine deaminase (E-ADA) in lymphocytes were determined. Furthermore, nucleotide and nucleoside levels as well as the secretion of pro- and anti-inflammatory cytokines were evaluated. Animals were treated with saline and quercetin in doses of 5, 25, and 50 mg/kg for 45 days. The result revealed that quercetin (50 mg/kg) reduced arthritis score and paw oedema, and increased the latency in the thermal hyperalgesia test. Histopathological analysis showed that all the doses of quercetin reduced infiltration of inflammatory cells. MPO activity was increased in the arthritis group; however, quercetin reduced this activity. E-NTPDase activity was increased in lymphocytes of arthritis rats, and treatment with quercetin reversed this increase. However, E-ADA activity was reduced in the arthritis group, and treatment with quercetin modulated the activity of this enzyme in arthritis rat groups. Serum adenosine levels were increased in arthritis, and the levels were lowered with quercetin treatment. Quercetin treatment in arthritis groups decreased the elevated levels of cytokines in the arthritis control group. Thus, quercetin demonstrated an anti-inflammatory effect, and this flavonoid may be a promising natural compound for the treatment of arthritis. SIGNIFICANCE OF THE STUDY: Quercetin may represent a potential therapeutic compound in the treatment of rheumatoid arthritis. Findings from this study indicate that quercetin suppresses swelling and attenuates the underlying inflammatory responses. This is the first report where quercetin was shown to modulate the immune response to arthritis via attenuation of the purinergic system (E-NTPDase and E-ADA activities) and the levels of IFN-gamma and IL-4. Thus, this work is relevant to basic research and may be translated into clinical practice.
Assuntos
AMP Desaminase/antagonistas & inibidores , Adenosina Trifosfatases/antagonistas & inibidores , Anti-Inflamatórios não Esteroides/farmacologia , Artrite Reumatoide/tratamento farmacológico , Citocinas/antagonistas & inibidores , Inibidores Enzimáticos/farmacologia , Quercetina/farmacologia , AMP Desaminase/metabolismo , Adenosina Trifosfatases/metabolismo , Animais , Artrite Reumatoide/induzido quimicamente , Artrite Reumatoide/metabolismo , Citocinas/metabolismo , Edema/induzido quimicamente , Edema/tratamento farmacológico , Edema/metabolismo , Feminino , Adjuvante de Freund , Ratos , Ratos WistarRESUMO
BACKGROUND: Quillaja saponaria Mol. bark contains a high concentration of triterpene saponins that have been used for centuries as a cleansing, antiinflammatory and analgesic agent in Chilean folk medicine. In earlier studies, in mice, both the anti-inflammatory as well as the antinociceptive effect of the major sapogenin, quillaic acid have been demonstrated (QA). OBJECTIVE: To determine the antihyperalgesic effect of QA one and seven days after itpl administration of complete Freund's adjuvant (CFA) in male mice using the hot plate test in the presence of complete Freund's adjuvant (HP/CFA) as an acute and chronic skeletal muscle pain model. METHODS: The present study evaluated the antihyperalgesic activity of QA against acute and chronic skeletal muscle pain models in mice using the hot plate test in the presence of complete Freund's adjuvant (HP/CFA), at 24 h (acute assay) and 7 days (chronic assay) , with dexketoprofen (DEX) as the reference drug. RESULTS: In acute and chronic skeletal muscle pain assays, QA at 30 mg/kg ip elicited its maximal antihyperalgesic effects (65.0% and 53.4%) at 24 h and 7 days, respectively. The maximal effect of DEX (99.0 and 94.1 at 24 h and 7 days, respectively) was induced at 100 mg/kg. CONCLUSION: QA and DEX elicit dose-dependent antihyperalgesic effects against acute and chronic skeletal muscle pain, but QA is more potent than DEX in the early and late periods of inflammatory pain induced by CFA.
Assuntos
Músculos Abdominais/efeitos dos fármacos , Hipoglicemiantes/farmacologia , Ácido Oleanólico/análogos & derivados , Dor/tratamento farmacológico , Quillaja/química , Animais , Hipoglicemiantes/química , Hipoglicemiantes/isolamento & purificação , Masculino , Camundongos , Camundongos Endogâmicos , Conformação Molecular , Ácido Oleanólico/química , Ácido Oleanólico/isolamento & purificação , Ácido Oleanólico/farmacologiaRESUMO
OBJECTIVES: To evaluate the analgesic effect of Glucantime (antimoniate N-methylglucamine) in Leishmania amazonensis infection and complete Freund's adjuvant (CFA), chronic paw inflammation model, in BALB/c mice. METHODS: Two models of chronic inflammatory pain in BALB/c mice paw were used: infection with L. amazonensis and CFA stimulation. Both animals models received daily treatment with Glucantime (10 mg/kg, i.p.) and during the treatment was measured the mechanical hyperalgesia with electronic version of von Frey filaments. After the treatment, the paw skin sample was collected for analysis of myeloperoxidase (MPO) and N-acetyl-ß-glucosaminidase (NAG) activity, and IL-1ß, TNF-α, IL-6, IFN-γ and IL-10 cytokines production by ELISA. KEY FINDINGS: Leishmania amazonensis-induced chronic inflammation with significant increase in mechanical hyperalgesia, MPO and NAG activity, and IL-1ß, TNF-α and IL-6 production in the paw skin. Glucantime (10 mg/kg, i.p.) inhibited L. amazonensis-induced mechanical hyperalgesia and IL-1ß and IL-6 cytokines productions. In chronic inflammatory model induced by CFA, Glucantime treatment during 7 days inhibited CFA-induced mechanical hyperalgesia, MPO and NAG activity, and IL-1ß, TNF-α, IL-6 and IFN-γ production as well as increased IL-10 production. CONCLUSIONS: Our data demonstrated that Glucantime reduced the chronic inflammatory pain induced by L. amazonensis and CFA stimuli by inhibiting the hyperalgesic cytokines production.
Assuntos
Dor Crônica/tratamento farmacológico , Inflamação/tratamento farmacológico , Leishmaniose Cutânea/tratamento farmacológico , Meglumina/uso terapêutico , Compostos Organometálicos/uso terapêutico , Acetilglucosaminidase/metabolismo , Animais , Dor Crônica/complicações , Citocinas/metabolismo , Adjuvante de Freund , Inflamação/induzido quimicamente , Inflamação/complicações , Masculino , Antimoniato de Meglumina , Camundongos , Peroxidase/metabolismo , Pele/metabolismoRESUMO
Recombinant simian IL-15 (siIL-15) was obtained for the preclinical assessment of an anti-human IL-15 vaccine. For this purpose, the cDNA from peripheral blood mononuclear cells of a Macaca fascicularis monkey was cloned into a pIL-2 vector. The siIL-15 was expressed in Escherichia coli strain W3110 as an insoluble protein which accounted for 13% of the total cellular proteins. Inclusion bodies were solubilized in an 8 M urea solution, which was purified by ion exchange and reverse phase chromatography up to 92% purity. The protein identity was validated by electrospray ionization-mass spectrometry, confirming the presence of the amino acids which distinguish the siIL-15 from human IL-15. The purified siIL-15 stimulates the proliferation of cytotoxic T-lymphocytes line (CTLL)-2 and Kit 225 cells with EC50 values of 3.1 and 32.5 ng/mL, respectively. Antisera from modified human IL-15-immunized macaques were reactive to human and simian IL-15 in enzyme-linked immunosorbent assays. Moreover, the anti-human IL-15 antibodies from immune sera inhibited siIL-15 activity in CTLL-2 and Kit 225 cells, supporting the activity and purity of recombinant siIL-15. These results indicate that the recombinant siIL-15 is biologically active in two IL-15-dependent cell lines, and it is also suitable for the preclinical evaluation of an IL-15-based therapeutic vaccine.
Assuntos
Interleucina-15/genética , Macaca fascicularis/genética , Vacinas Sintéticas/genética , Animais , Linhagem Celular , Clonagem Molecular/métodos , Escherichia coli/genética , Humanos , Interleucina-15/imunologia , Macaca fascicularis/imunologia , Camundongos , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Linfócitos T Citotóxicos/imunologia , Vacinas Sintéticas/imunologiaRESUMO
ABSTRACT This study assesses the anti-arthritic effect of the affinin-enriched (spilanthol, main alkamide) hexane extract from the roots of Heliopsis longipes (A. Gray) S.F. Blake, Asteraceae, on a Freund adjuvant-induced arthritis model in rodents. The extract was orally administered at a dose of 2, 6.6, or 20 mg/kg; a significant edema-inhibitory activity in the acute and chronic phases was observed with a dose of 2 and 20 mg/kg, respectively. The extract showed higher anti-inflammatory and anti-arthritic effects than the reference drug phenylbutazone (80 mg/kg). Moreover, the extract prevented the occurrence of secondary lesions associated to this pharmacological model.
RESUMO
La artritis reumatoide (AR) es una enfermedad crónica de naturaleza autoinmune e inflamatoria que conduce a la formación de pannus seguido de la destrucción de las articulaciones, se caracteriza por hiperplasia sinovial, inflamación y angiogénesis. La especie vegetal Baccharis latifolia es utilizada tradicionalmente en muchas regiones de nuestro país para tratar el dolor, la inflamación y la artritis. En el presente estudio se evaluó la actividad antiartrítica del extracto etanólico de B. latifolia en modelos murinos de artritis reumatoide inducida por adyuvante, la estimación del edema/espesor de la pata inflamada, parámetros hematológicos (hemoglobina, velocidad de sedimentación globular, recuento de eritrocitos, recuento total de leucocitos) y observación radiológica fueron evaluados. La administración oral del extracto etanólico de B. latifolia (600 mg/kg de p.c.) inhibió significativamente (p<0.001) el incremento del edema/espesor de la pata en el modelo de artritis subcrónica. Del mismo modo, B. latifolia (500 mg/kg de p.c.) inhibió significativamente el incremento del edema/espesor de la pata en el modelo de artritis crónica (p<0.05, p<0.01), los pesos de los animales se mantuvieron sin variación durante el tratamiento. Por otro lado, los parámetros hematológicos señalan que los niveles de hemoglobina disminuyen en ratones artríticos y que esta disminución es revertida tras la administración de los extractos de B. latifolia, este mismo perfil de recuperación es observado tras el recuento de glóbulos rojos. Adicionalmente, la velocidad de sedimentación globular (VSG) incrementada en ratones artríticos, es revertida tras la administración de B. latifolia. El análisis radiológico evidenció el efecto del extracto etanólico de B. latifolia en el retraso de la destrucción ósea. Los resultados sugieren que el extracto etanólico de B. latifolia tiene una potencial actividad antiartrítica.
Rheumatoid arthritis (RA) is a chronic autoimmune and inflammatory disease leading to pannus formation followed by the destruction of the joints; it is characterized by synovial hyperplasia, inflammation and angiogenesis. The plant species Baccharis latifolia is traditionally used in many regions of our country to treat pain, inflammation and arthritis. In the present study the anti-arthritic activity of ethanol extract of B. latifolia was evaluated in murine experimental model of rheumatoid arthritis induced by adjuvant, edema estimation / thickness of the inflamed foot, hematological parameters (hemoglobin, erythrocyte sedimentation rate, erythrocyte count was assessed, total white blood cell) and radiological observation were evaluated. The oral administration of the ethanolic extract of B. latifolia (600 mg / kg p.c.) significantly inhibited (p<0,001) increased edema / paw thickness in the subchronic model of arthritis. Similarly, B. latifolia (500 mg / kg bw) significantly inhibited the increase of edema / paw thickness in the model of chronic arthritis (p <0.05, p <0.01), the weights of the animals were kept without variation during treatment. Moreover hematological parameters indicate that hemoglobin levels decrease in arthritic mice and that this decline is reversed after administration of the extracts of B. latifolia, this same profile is observed recovery after red blood cell count. Additionally, erythrocyte sedimentation rate (ESR) increased in arthritic mice is reversed after administration of B. latifolia. The radiological analysis showed the effect of ethanol extract of B. latifolia in delaying bone destruction. The results suggest that the ethanolic extract of B. latifolia has potential antiarthritic activity.
Assuntos
Artrite Reumatoide , Doença Crônica , Baccharis , Dor , Sedimentação Sanguínea , Hemoglobinas , Etanol , Padrões de Referência , Inflamação , LeucócitosRESUMO
Hypoferremia observed during systemic inflammatory disorders is regulated by hepcidin. Hepcidin up-regulation is particularly important during acute inflammation, as it restricts the availability of iron, which is necessary for pathogenic microorganism growth before adaptive immunity occurs. The aim of this study was to evaluate the clinical findings and hepatic hepcidin mRNA expression in horses using a Freund's complete adjuvant (FCA) model of inflammation. The expression of hepcidin mRNA in the liver was determined in healthy horses following two intramuscular injections of FCA at 0 h and 12 h. Plasma iron and fibrinogen concentrations were measured at multiple time points between 0 h and 240 h post-FCA injection (PI). Hepcidin mRNA expression was determined by RT-qPCR using liver biopsy samples performed at 0 h (control), 6 h and 18 h PI. The mean plasma fibrinogen level was significantly different from the control values only between 120 and 216 h PI. The mean plasma iron level was significantly lower than the control between 16 and 72 h PI, reaching the lowest levels at 30 h PI (33 % of the initial value), and returned to the reference value from 96 h PI to the end of the experiment. Hepcidin mRNA expression increased at 6 h PI and remained high at 18 h PI. The iron plasma concentration was an earlier indicator of inflammatory processes in horses when compared with fibrinogen and might be useful for the early detection of inflammation in the horse. FCA administration caused the rapid onset of hypoferremia, and this effect was likely the result of up-regulated hepatic hepcidin gene expression. This study emphasizes the importance of hepcidin and iron metabolism during inflammation in horses.
A hipoferremia observada durante os processos inflamatórios sistêmicos é mediada pela hepcidina. O aumento da expressão da hepcidina é particularmente importante durante a inflamação aguda, por restringir a disponibilidade de ferro necessária para o crescimento de microrganismos patogênicos antes que a imunidade adaptativa ocorra. O objetivo deste estudo foi avaliar os achados clínicos e a expressão hepática do RNA mensageiro (RNAm) da hepcidina em cavalos após a indução da inflamação com Adjuvante completo de Freund (FCA). A expressão hepática do RNAm da hepcidina foi determinada em cavalos sadios após duas administrações intramusculares de FCA às 0 h (M0) e 12 h (M12). As concentrações plasmáticas de ferro e fibrinogênio foram mensuradas em múltiplos momentos entre 0 h e 240 h (M240) após a primeira administração de FCA (PI). A expressão do RNAm da hepcidina foi determinada por RT-qPCR usando amostras de biopsias hepáticas colhidas as 0 h (controle), 6 h (M6) e 18 h (M18) PI. A concentração plasmática média de fibrinogênio foi estatisticamente diferente do M0 entre 120 h e 216 h PI. A concentração plasmática média de ferro foi significantemente menor que o controle entre 16 h e 72 h PI, alcançou o nível mais baixo às 30 h PI (33% do valor inicial) e retornou aos valores de referência entre 96 h PI e até o final do experimento. A expressão do RNAm da hepcidina aumentou no M6 e permaneceu alta no M18. A concentração plasmática de ferro foi um indicador precoce da inflamação quando comparada com o fibrinogênio e pode ser útil na detecção precoce da inflamação em cavalos. A administração do FCA causou um rápido início da hipoferremia, e isto foi resultante do aumento da expressão hepática da hepcidina. Estes resultados enfatizam a importância da hepcidina e do metabolismo do ferro durante a inflamação em cavalos.
Assuntos
Animais , Cavalos/metabolismo , Deficiências de Ferro/diagnóstico , Fibrinogênio/análise , Hepcidinas/análise , Adjuvante de Freund , Inflamação/veterináriaRESUMO
Hypoferremia observed during systemic inflammatory disorders is regulated by hepcidin. Hepcidin up-regulation is particularly important during acute inflammation, as it restricts the availability of iron, which is necessary for pathogenic microorganism growth before adaptive immunity occurs. The aim of this study was to evaluate the clinical findings and hepatic hepcidin mRNA expression in horses using a Freund's complete adjuvant (FCA) model of inflammation. The expression of hepcidin mRNA in the liver was determined in healthy horses following two intramuscular injections of FCA at 0 h and 12 h. Plasma iron and fibrinogen concentrations were measured at multiple time points between 0 h and 240 h post-FCA injection (PI). Hepcidin mRNA expression was determined by RT-qPCR using liver biopsy samples performed at 0 h (control), 6 h and 18 h PI. The mean plasma fibrinogen level was significantly different from the control values only between 120 and 216 h PI. The mean plasma iron level was significantly lower than the control between 16 and 72 h PI, reaching the lowest levels at 30 h PI (33 % of the initial value), and returned to the reference value from 96 h PI to the end of the experiment. Hepcidin mRNA expression increased at 6 h PI and remained high at 18 h PI. The iron plasma concentration was an earlier indicator of inflammatory processes in horses when compared with fibrinogen and might be useful for the early detection of inflammation in the horse. FCA administration caused the rapid onset of hypoferremia, and this effect was likely the result of up-regulated hepatic hepcidin gene expression. This study emphasizes the importance of hepcidin and iron metabolism during inflammation in horses.(AU)
A hipoferremia observada durante os processos inflamatórios sistêmicos é mediada pela hepcidina. O aumento da expressão da hepcidina é particularmente importante durante a inflamação aguda, por restringir a disponibilidade de ferro necessária para o crescimento de microrganismos patogênicos antes que a imunidade adaptativa ocorra. O objetivo deste estudo foi avaliar os achados clínicos e a expressão hepática do RNA mensageiro (RNAm) da hepcidina em cavalos após a indução da inflamação com Adjuvante completo de Freund (FCA). A expressão hepática do RNAm da hepcidina foi determinada em cavalos sadios após duas administrações intramusculares de FCA às 0 h (M0) e 12 h (M12). As concentrações plasmáticas de ferro e fibrinogênio foram mensuradas em múltiplos momentos entre 0 h e 240 h (M240) após a primeira administração de FCA (PI). A expressão do RNAm da hepcidina foi determinada por RT-qPCR usando amostras de biopsias hepáticas colhidas as 0 h (controle), 6 h (M6) e 18 h (M18) PI. A concentração plasmática média de fibrinogênio foi estatisticamente diferente do M0 entre 120 h e 216 h PI. A concentração plasmática média de ferro foi significantemente menor que o controle entre 16 h e 72 h PI, alcançou o nível mais baixo às 30 h PI (33% do valor inicial) e retornou aos valores de referência entre 96 h PI e até o final do experimento. A expressão do RNAm da hepcidina aumentou no M6 e permaneceu alta no M18. A concentração plasmática de ferro foi um indicador precoce da inflamação quando comparada com o fibrinogênio e pode ser útil na detecção precoce da inflamação em cavalos. A administração do FCA causou um rápido início da hipoferremia, e isto foi resultante do aumento da expressão hepática da hepcidina. Estes resultados enfatizam a importância da hepcidina e do metabolismo do ferro durante a inflamação em cavalos.(AU)