RESUMO
Voltage-sensitive calcium channels (VSCCs) underlie cell excitability and are involved in the mechanisms that generate and maintain neuropathic and inflammatory pain. We evaluated in rats the effects of two VSCC blockers, ω-conotoxin MVIIA and Phα1ß, in models of inflammatory and neuropathic pain induced with complete Freund's adjuvant (CFA) and chronic constrictive injury (CCI), respectively. We also evaluated the effects of the toxins on capsaicin-induced Ca(2+) influx in dorsal root ganglion (DRG) neurons obtained from rats exposed to both models of pain. A single intrathecal injection of Phα1ß reversibly inhibits CFA and CCI-induced mechanical hyperalgesia longer than a single injection of ω-conotoxin MVIIA. Phα1ß and MVIIA also inhibited capsaicin-induced Ca(2+) influx in DRG neurons. The inhibitory effect of Phα1ß on capsaicin-induced calcium transients in DRG neurons was greater in the CFA model of pain, while the inhibitory effect of ω-conotoxin MVIIA was greater in the CCI model. The management of chronic inflammatory and neuropathic pain is still a major challenge for clinicians. Phα1ß, a reversible inhibitor of VSCCs with a preference for N-type Ca(2+) channels, has potential as a novel therapeutic agent for inflammatory and neuropathic pain. Clinical studies are necessary to establish the role of Phα1ß in the treatment of chronic pain.
Assuntos
Analgésicos/uso terapêutico , Modelos Animais de Doenças , Neuralgia/tratamento farmacológico , Neurotoxinas/uso terapêutico , Venenos de Aranha/uso terapêutico , Aranhas , ômega-Conotoxinas/uso terapêutico , Analgésicos/isolamento & purificação , Animais , Células Cultivadas , Avaliação Pré-Clínica de Medicamentos/métodos , Inflamação/tratamento farmacológico , Inflamação/patologia , Masculino , Neuralgia/patologia , Neurotoxinas/isolamento & purificação , Ratos , Ratos Wistar , Caramujos , Venenos de Aranha/isolamento & purificação , ômega-Conotoxinas/isolamento & purificaçãoRESUMO
This study aimed to investigate the feasibility of preparing a hydrogel based on (OMePhSe)2-loaded poly(Æ-caprolactone) nanocapsules using carrageenan gum as a gel-forming agent. Furthermore, the anti-inflammatory action of hydrogel was assessed in an animal model of skin lesion induced by ultraviolet B (UVB) radiation in mice. Nanocapsules were prepared using the interfacial deposition of preformed polymer technique. The hydrogels were obtained by the direct addition of nanocapsules suspension in carrageenan gum (3%). Formulations with free compound, vehicle, and blank nanocapsules were also produced. The hydrogels were characterized by pH, compound content, diameter, spreadability, rheological behavior, and permeation profile. The pharmacological performance was assessed in an animal model of skin injury induced by UVB-radiation in male Swiss mice. All hydrogels had pH around 7.0, compound content close to the theoretical value (2.5 mg/g), an average diameter in nanometric range (around 350 nm), non-Newtonian flow with pseudoplastic behavior, and suitable spreadability factor. The nano-based hydrogel increased the compound content in the epidermis and dermis layers in comparison to the formulation prepared with non-encapsulated (OMePhSe)2. Stability studies revealed that the hydrogels of nanoencapsulated compound had superior physicochemical stability in comparison to the formulation of free (OMePhSe)2. Moreover, topical treatment with the hydrogel containing (OMePhSe)2 loaded-nanocapsules was more effective in reducing ear thickness and the inflammatory process induced by UVB radiation in mice. Herein, a polysaccharide was applied as a gel-forming agent using a simple and low-cost method. Besides, a superior permeation profile and improved pharmacological action were achieved by the compound encapsulation.
Assuntos
Nanocápsulas , Compostos Organosselênicos , Animais , Anti-Inflamatórios/uso terapêutico , Carragenina , Hidrogéis , Masculino , CamundongosRESUMO
Beta-carotene is a carotenoid precursor of vitamin A, known for its biological activities. Due to its high hydrophobicity, nanonization processes, i.e. the transformation into nanoparticles, can improve its water affinity, and therefore the activity in aqueous systems. The objective of this study was to produce beta-carotene nanoparticles by the solid dispersion method and to evaluate their effects on the activity of glutathione-S-transferase and acetylcholinesterase enzymes using Drosophila melanogaster (DM) homogenate, the superoxide dismutase- and catalase-like activities under in vitro conditions, and their cytotoxic properties against tumor and non-tumor cells. The formed nanometric beta-carotene particles resulted in stable colloids, readily dispersed in water, able to modulate acetylcholinesterase (AChE) activity, and presenting high potential to control the cholinergic system. Beta-carotene nanoparticles, at concentrations much lower than the pure pristine beta-carotene, presented in vitro mimetic activity to superoxide dismutase and altered glutathione-S-transferase activity in DM tissue. The content of hydrogen peroxide was neither affected by the nanoparticles (in aqueous solution) nor by pristine beta-carotene (in DMSO). In the cytotoxic assays, beta-carotene nanoparticles dispersed in water showed activity against four different tumor cell lines. Overall, beta-carotene nanoparticles presented significant bioactivity in aqueous medium surpassing their high hydrophobicity constraint.
Assuntos
Antioxidantes/farmacologia , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/efeitos dos fármacos , Drosophila melanogaster/enzimologia , beta Caroteno/farmacologia , Animais , Antioxidantes/química , Catalase/metabolismo , Linhagem Celular Tumoral , Drosophila melanogaster/genética , Glutationa Peroxidase/metabolismo , Humanos , Interações Hidrofóbicas e Hidrofílicas , Nanopartículas/química , Superóxido Dismutase/metabolismo , beta Caroteno/químicaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: The medicinal plant generally known as monkey's comb (Amphilophium crucigerum) has been popularly described for the treatment of neuropathic and inflammatory pain, specially seeds preparations. AIM OF THE STUDY: The goal of the present study was to evaluate the antinociceptive effect of the crude extract (Crd) and dichloromethane fraction (Dcm) of A. crucigerum seeds, and investigate the involvement of transient receptor potential vanilloid 1 (TRPV1) receptor in this effect. MATERIALS AND METHODS: Male Swiss mice were used in this study. The effects of Crd and Dcm was tested on capsaicin-induced Ca2+ influx or the specific binding of [3H]-resiniferatoxin. Moreover, after treatment with Crd or Dcm, animals were exposed to acute pain (hot water tail-flick and capsaicin intraplantar test) or chronic pain models (injection of complete Freund's adjuvant or partial ligation of the sciatic nerve). Acute adverse effects were also noted: locomotor activity, corporal temperature, hepatic or renal damage, gastrointestinal transit alteration, and ulcerogenic activity. RESULTS: The oral administration of Crd or Dcm resulted in an antinociceptive effect in the hot water tail-flick (48°C) and capsaicin intraplantar tests. Furthermore, these preparations exhibited antinociceptive and anti-inflammatory effects in a chronic inflammatory pain model, and antinociceptive effects in a neuropathic pain model. Moreover, Crd and Dcm reduced capsaicin-induced Ca2+ influx and diminished the [3H]-resiniferatoxin specific binding to spinal cord membranes. Acute adverse events were not found with Crd or Dcm administration. CONCLUSION: In conclusion, our results support the analgesic effect of A. crucigerum and suggest the presence of compounds that may act as TRPV1 antagonists.
Assuntos
Analgésicos/farmacologia , Bignoniaceae/química , Dor Crônica/prevenção & controle , Etanol/química , Cloreto de Metileno/química , Neuralgia/prevenção & controle , Nociceptividade/efeitos dos fármacos , Dor Nociceptiva/prevenção & controle , Extratos Vegetais/farmacologia , Sementes/química , Solventes/química , Medula Espinal/efeitos dos fármacos , Canais de Cátion TRPV/antagonistas & inibidores , Analgésicos/isolamento & purificação , Analgésicos/metabolismo , Analgésicos/toxicidade , Animais , Ligação Competitiva , Sinalização do Cálcio/efeitos dos fármacos , Capsaicina/farmacologia , Dor Crônica/metabolismo , Dor Crônica/fisiopatologia , Modelos Animais de Doenças , Diterpenos/metabolismo , Relação Dose-Resposta a Droga , Masculino , Camundongos , Neuralgia/metabolismo , Neuralgia/fisiopatologia , Dor Nociceptiva/metabolismo , Dor Nociceptiva/fisiopatologia , Medição da Dor , Fitoterapia , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/metabolismo , Extratos Vegetais/toxicidade , Plantas Medicinais , Ligação Proteica , Transdução de Sinais/efeitos dos fármacos , Medula Espinal/metabolismo , Medula Espinal/fisiopatologia , Sinaptossomos/efeitos dos fármacos , Sinaptossomos/metabolismo , Canais de Cátion TRPV/metabolismoRESUMO
BACKGROUND: Atherosclerosis is an inflammatory disease that affects the arterial wall leading to myocardial, cerebral, and peripheral ischemic syndromes. The use of low doses of aspirin inhibits platelet aggregation and inflammation and prevents cardiovascular mortality. However, ASA may produce hemorrhagic events. Thus, several studies have sought new natural compounds to suppress platelet aggregation without causing serious adverse effects. PURPOSE: In this sense, this study aims to compare the effects of Campomanesia xanthocarpa plant extract with those of acetylsalicylic acid (ASA) on inflammatory parameters observed in homozygous mice knockout for the low-density lipoprotein receptor (LDLr-KO) treated with a hypercholesterolemic diet. MATERIAL AND METHODS: In this study, 28 male LDLr-KO mice were divided into three groups and fed a hypercholesterolemic diet for 4 weeks. Thereafter, the animals that received the hypercholesterolemic diet were treated for 5 days with (1) distilled water, (2) C. xanthocarpa extract, or (3) acetylsalicylic acid. The levels of inflammatory markers were assessed in the blood samples. The gastric tolerability of the animals after oral administration of the treatments was assessed through quantification of the lesions in the gastric mucosa. RESULTS: The levels of proinflammatory cytokines IL-1, IL-6, TNF-α, and INF-γ were reduced to 19.2 ± 3%, 20.4 + 1.3%, 24.7 ± 1.2%, and 20.8 ± 1.7%, respectively, in the group treated with C. xanthocarpa, when compared to control group. Furthermore, treatment with plant extract significantly increased the levels of the anti-inflammatory cytokine IL-10 by 27.3 ± 5.9%, but ASA showed no significant effect on the same cytokines when compared to the control group, with the exception of IL-10, which presented an increase of 8.6 ± 3.5%. Treatments with C. xanthocarpa and ASA also caused significant reductions of 26.4 ± 3% and 38.4± 6% in the serum levels of oxLDL, respectively. However, only treatment with C. xanthocarpa reduced the levels of anti-oxLDL antibodies when compared with the control (25.8 ± 6%). In addition, the analyzed extract did not induce ulcerogenic activity, while ASA induced the formation of lesions. CONCLUSION: In conclusion, treatment with C. xanthocarpa causes anti-inflammatory activity in hypercholesterolemic animals, with results superior to those obtained with the use of ASA.
Assuntos
Aspirina/uso terapêutico , Aterosclerose/tratamento farmacológico , Hipercolesterolemia/tratamento farmacológico , Inflamação/tratamento farmacológico , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Animais , Aspirina/farmacologia , Brasil , Lipoproteínas LDL/sangue , Lipoproteínas LDL/efeitos dos fármacos , Masculino , Camundongos , Camundongos Knockout , Myrtaceae/química , Estresse Oxidativo/efeitos dos fármacos , Plantas Medicinais/químicaRESUMO
Excessive UV-B radiation by sunlight produces inflammatory and oxidative damage of skin, which can lead to sunburn, photoaging, and cancer. This study evaluated whether nanoencapsulation improves the protective effects of rice bran oil against UVB radiation-induced skin damage in mice. Lipid-core nanocapsules containing rice bran oil were prepared, and had mean size around 200 nm, negative zeta potential (â¼-9 mV), and low polydispersity index (<0.20). In order to allow application on the skin, a hydrogel containing the nanoencapsulated rice bran oil was prepared. This formulation was able to prevent ear edema induced by UVB irradiation by 60 ± 9%, when compared with a hydrogel containing LNC prepared with a mixture of medium chain triglycerides instead of rice bran oil. Protein carbonylation levels (biomarker of oxidative stress) and NF-κB nuclear translocation (biomarker of pro-inflammatory and carcinogenesis response) were reduced (81% and 87%, respectively) in animals treated with the hydrogel containing the nanoencapsulated rice bran oil. These in vivo results demonstrate the beneficial effects of nanoencapsulation to improve the protective properties of rice bran oil on skin damage caused by UVB exposure.
Assuntos
Anti-Inflamatórios/administração & dosagem , Antioxidantes/administração & dosagem , Portadores de Fármacos , Edema/prevenção & controle , Nanopartículas , Óleos de Plantas/administração & dosagem , Pele/efeitos dos fármacos , Queimadura Solar/prevenção & controle , Raios Ultravioleta , Animais , Anti-Inflamatórios/química , Antioxidantes/química , Química Farmacêutica , Citoproteção , Modelos Animais de Doenças , Edema/metabolismo , Edema/patologia , Hidrogéis , Mediadores da Inflamação/metabolismo , Masculino , Camundongos , NF-kappa B/metabolismo , Nanomedicina , Estresse Oxidativo/efeitos dos fármacos , Tamanho da Partícula , Óleos de Plantas/química , Carbonilação Proteica/efeitos dos fármacos , Óleo de Farelo de Arroz , Pele/metabolismo , Pele/patologia , Queimadura Solar/metabolismo , Queimadura Solar/patologia , Tecnologia Farmacêutica/métodosRESUMO
Administration of the compound triterpene 3ß, 6ß, 16ß-trihidroxilup-20(29)-ene (TTHL) resulted in antinociceptive activity in several pain models in mice. Because pain and epilepsy have common mechanisms, and several anticonvulsants are clinically used to treat painful disorders, we investigated the anticonvulsant potential of TTHL. Behavioral and electrographic recordings revealed that pretreatment with TTHL (30 mg/kg; i.g.) increased the latencies to the first clonic seizure to the tonic-clonic and reduced the duration of the generalized seizures induced by the GABA(A) receptor antagonist PTZ (80 g; i.p.). The TTHL pretreatment also protected against PTZ-induced deleterious effects, as characterized by protein carbonylation, lipid peroxidation, [(3)H] glutamate uptake and the inhibition of Na(+),K(+)-ATPase (subunits α(1) and α(2)/α(3)). Although TTHL did not exhibit DPPH, ABTS radical scavenging activity per se and does not alter the binding of [(3)H]flunitrazepam to the benzodiazepinic site of the GABA(A) receptor, this compound was effective in preventing behavioral and EEG seizures, as well as the inhibition of Na(+),K(+)-ATPase induced by ouabain. These results suggest that the protection against PTZ-induced seizures elicited by TTHL is due to Na(+),K(+)-ATPase activity maintenance. In fact, experiments in homogenates of the cerebral cortex revealed that PTZ (10 mM) reduced Na(+),K(+)-ATPase activity and that previous incubation with TTHL (10 µM) protected against this inhibition. Collectively, these data indicate that the protection exerted by TTHL in this model of convulsion is not related to antioxidant activity or GABAergic activity. However, these results demonstrated that the effective protection of Na(+),K(+)-ATPase elicited by this compound protects against the damage due to neuronal excitability and oxidation that is induced by PTZ.
Assuntos
Fármacos Neuroprotetores/administração & dosagem , Estresse Oxidativo/fisiologia , Pentilenotetrazol/toxicidade , Convulsões/enzimologia , Convulsões/prevenção & controle , ATPase Trocadora de Sódio-Potássio/fisiologia , Triterpenos/administração & dosagem , Animais , Ativação Enzimática/efeitos dos fármacos , Ativação Enzimática/fisiologia , Injeções Intraventriculares , Camundongos , Fármacos Neuroprotetores/química , Estresse Oxidativo/efeitos dos fármacos , Convulsões/induzido quimicamente , Triterpenos/químicaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Achillea millefolium L. (Asteraceae), known as yarrow ("mil folhas"), has been used as folk medicine for gastrointestinal disorders, inflammation, anxiety, and insomnia. AIM: To evaluate the potential anxiolytic-like effect of hydroalcoholic extract of Achillea millefolium L. in animal models. METHODS: The present study evaluated the effects of the hydroalcoholic extract from the aerial parts of Achillea millefolium L. in mice subjected to the elevated plus-maze, marble-burying, and open-field tests. Additionally, the GABA(A)/benzodiazepine (BDZ) mediation of the effects of Achillea millefolium was evaluated by pretreatment with the noncompetitive GABA(A) receptor antagonist picrotoxin and the BDZ antagonist flumazenil and by [(3)H]-flunitrazepam binding to the BDZ site on the GABA(A) receptor. RESULTS: Achillea millefolium exerted anxiolytic-like effects in the elevated plus-maze and marble-burying test after acute and chronic (25 days) administration at doses that did not alter locomotor activity. This behavioral profile was similar to diazepam. The effects of Achillea millefolium in the elevated plus-maze were not altered by picrotoxin pretreatment but were partially blocked by flumazenil. Furthermore, Achillea millefolium did not induce any changes in [(3)H]-flunitrazepam binding. CONCLUSION: The results indicate that the orally administered hydroalcoholic extract of Achillea millefolium L. exerted anxiolytic-like effects that likely were not mediated by GABA(A)/BDZ neurotransmission and did not present tolerance after short-term, repeated administration.
Assuntos
Achillea , Ansiolíticos/uso terapêutico , Ansiedade/tratamento farmacológico , Comportamento Animal/efeitos dos fármacos , Fitoterapia , Extratos Vegetais/uso terapêutico , Animais , Ansiolíticos/farmacologia , Benzodiazepinas/farmacologia , Modelos Animais de Doenças , GABAérgicos/farmacologia , Locomoção/efeitos dos fármacos , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Medicina Tradicional , Camundongos , Camundongos Endogâmicos , Componentes Aéreos da Planta , Extratos Vegetais/farmacologiaRESUMO
Gout is characterized by the deposition of monosodium urate (MSU) crystals. Despite being one of the most painful forms of arthritis, gout and the mechanisms responsible for its acute attacks are poorly understood. In the present study, we found that MSU caused dose-related nociception (ED(50) [ie, the necessary dose of MSU to elicit 50% of the response relative to the control value]=0.04 [95% confidence interval 0.01-0.11]mg/paw) and edema (ED(50)=0.08 [95% confidence interval 0.04-0.16]mg/paw) when injected into the hind paw of rats. Treatment with the selective TRPV1 receptor (also known as capsaicin receptor and vanilloid receptor-1) antagonists SB366791 or AMG9810 largely prevented nociceptive and edematogenic responses to MSU. Moreover, the desensitization of capsaicin-sensitive afferent fibers as well as pretreatment with the tachykinin NK(1) receptor antagonist RP 67580 also significantly prevented MSU-induced nociception and edema. Once MSU was found to induce mast cell stimulation, we investigated the participation of these cells on MSU effects. Prior degranulation of mast cells by repeated treatment with the compound 48/80 decreased MSU-induced nociception and edema or histamine and serotonin levels in the injected tissue. Moreover, pretreatment with the mast cell membrane stabilizer cromolyn effectively prevented nociceptive and edematogenic responses to MSU. MSU induced a release of histamine, serotonin, and tryptase in the injected tissue, confirming mast cell degranulation. Furthermore, the antagonism of histaminergic H1 and serotoninergic receptors decreased the edema, but not the nociception of MSU. Finally, the prevention of the tryptase activity was capable of largely reducing both MSU-induced nociception and edema. Collectively, the present findings demonstrate that MSU produces nociceptive and edematogenic responses mediated by TRPV1 receptor activation and mast cell degranulation.
Assuntos
Antioxidantes/efeitos adversos , Edema/induzido quimicamente , Dor/induzido quimicamente , Canais de Cátion TRPV/metabolismo , Ácido Úrico/efeitos adversos , Acrilamidas/uso terapêutico , Anilidas/uso terapêutico , Animais , Antialérgicos/uso terapêutico , Antiasmáticos/administração & dosagem , Compostos Bicíclicos Heterocíclicos com Pontes/uso terapêutico , Capsaicina/administração & dosagem , Cinamatos/uso terapêutico , Cromolina Sódica/administração & dosagem , Modelos Animais de Doenças , Diterpenos/farmacocinética , Edema/tratamento farmacológico , Edema/metabolismo , Edema/prevenção & controle , Gabexato/uso terapêutico , Histamina/metabolismo , Masculino , Mastócitos/efeitos dos fármacos , Metisergida/administração & dosagem , Dor/tratamento farmacológico , Dor/metabolismo , Dor/prevenção & controle , Prometazina/uso terapêutico , Prostaglandina-Endoperóxido Sintases/metabolismo , Ligação Proteica/efeitos dos fármacos , Ratos , Ratos Wistar , Inibidores de Serina Proteinase/administração & dosagem , Serotonina/metabolismo , Antagonistas da Serotonina/administração & dosagem , Trítio/farmacocinéticaRESUMO
Polyamines (putrescine, spermidine and spermine) are important endogenous regulators of ion channels, such as vanilloid (TRPV1), glutamatergic (NMDA or AMPA/kainate) and acid-sensitive (ASIC) receptors. In the present study, we have investigated the possible nociceptive effect induced by polyamines and the mechanisms involved in this nociception in vivo. The subcutaneous (s.c.) injection of capsaicin (as positive control), spermine, spermidine or putrescine produced nociception with ED(50) of 0.16 (0.07-0.39)nmol/paw, 0.4 (0.2-0.7) µmol/paw, 0.3 (0.1-0.9) µmol/paw and 3.2 (0.9-11.5) µmol/paw, respectively. The antagonists of NMDA (MK801, 1 nmol/paw), AMPA/kainate (DNQX, 1 nmol/paw) or ASIC receptors (amiloride, 100 nmol/paw) failed to reduce the spermine-trigged nociception. However, the TRPV1 antagonists capsazepine or SB366791 (1 nmol/paw) reduced spermine-induced nociception, with inhibition of 81 ± 10 and 68 ± 9%, respectively. The previous desensitization with resiniferatoxin (RTX) largely reduced the spermine-induced nociception and TRPV1 expression in the sciatic nerve, with reductions of 82 ± 9% and 67 ± 11%, respectively. Furthermore, the combination of spermine (100 nmol/paw) and RTX (0.005 fmol/paw), in doses which alone were not capable of inducing nociception, produced nociceptive behaviors. Moreover, different concentrations of spermine (3-300 µM) enhanced the specific binding of [(3)H]-RTX to TRPV1 receptor. Altogether, polyamines produce spontaneous nociceptive effect through the stimulation of TRPV1, but not of ionotropic glutamate or ASIC receptors.
Assuntos
Nociceptores/efeitos dos fármacos , Espermina/farmacologia , Canais de Cátion TRPV/efeitos dos fármacos , Canais Iônicos Sensíveis a Ácido , Animais , Poliaminas Biogênicas/farmacologia , Western Blotting , Diterpenos/metabolismo , Diterpenos/farmacologia , Ácido Glutâmico/farmacologia , Masculino , Camundongos , Fibras Nervosas/efeitos dos fármacos , Proteínas do Tecido Nervoso/efeitos dos fármacos , Dor/induzido quimicamente , Dor/prevenção & controle , Medição da Dor/efeitos dos fármacos , Receptores de AMPA/efeitos dos fármacos , Receptores de Ácido Caínico/efeitos dos fármacos , Receptores de N-Metil-D-Aspartato/efeitos dos fármacos , Canais de Sódio/efeitos dos fármacos , Espermina/administração & dosagemRESUMO
The kallikrein-kinin system (KKS) has been described as an important mediator of physiologic processes. Kallikreins use kininogen (KNG) as substrate to generate bradykinin, the main active peptide of the KKS that acts through two types of receptors, the B(1)R and the B(2)R. The goal of this study was to characterize some components of the KKS in different compartments of the ovary during the bovine ovulation process. The KNG, B(1)R and B(2)R mRNA expression patterns were assessed in theca and granulosa cells, as well as the bradykinin concentration and kallikrein-like activity in follicular fluid of bovine periovulatory follicles. To obtain a periovulatory follicle (≥12 mm), twenty-seven cows were submitted to estrus synchronization protocol and ovariectomized by colpotomy at 0, 3, 6, 12 or 24h after a GnRH-analog injection (gonadorelin; 100 µg, IM). Follicular fluid was aspirated for enzymatic assays while granulosa and theca cells were harvested for mRNA analysis. The mRNA expressions in follicular cells were evaluated by real-time RT-PCR and data representation related to the cyclophilin housekeeping gene. The bradykinin concentration and kallikrein-like activity were measured in follicular fluid by enzymatic immunoassay and selective substrate cleavage, respectively. The B(2)R expression in theca cells and B(1)R expression in theca and granulosa cells showed different profiles during the periovulatory period (P<0.05). The bradykinin concentration and kallikrein-like activity in the follicular fluid were different (P<0.05) due to the time during the ovulation process. KNG mRNA expression was similar for both follicular cell types (P>0.05). Taken together, these results provide an important characterization of the presence and possible KKS regulation during the bovine ovulation.