RESUMEN
Melanoma is a very aggressive tumor that arises from melanocytes. Late stage and widely spread diseases do not respond to standard therapeutic approaches. The kallikrein-kinin system (KKS) participates in biological processes such as vasodilatation, pain and inflammatory response. However, the role of KKS in tumor formation and progression is not completely understood. The role of the host kinin B1 receptor in melanoma development was evaluated using a syngeneic melanoma model. Primary tumors and metastasis were respectively induced by injecting B16F10 melanoma cells, which are derived from C57BL/6 mice, subcutaneously or in the tail vein in wild type C57BL/6 and B1 receptor knockout mice (B1(-/-)). Tumors developed in B1(-/-) mice presented unfavorable prognostic factors such as increased incidence of ulceration, higher levels of IL-10, higher activation of proliferative pathways such as ERK1/2 and Akt, and increased mitotic index. Furthermore, in the metastasis model, B1(-/-) mice developed larger metastatic colonies in the lung and lower CD8(+)immune effector cells when compared with WT animals. Altogether, our results provide evidences that B1(-/-) animals developed primary tumors with multiple features associated with poor prognosis and unfavorable metastatic onset, indicating that the B1 receptor may contribute to improve the host response against melanoma progression.
Asunto(s)
Regulación Neoplásica de la Expresión Génica , Neoplasias Pulmonares/genética , Melanoma Experimental/genética , Receptor de Bradiquinina B1/genética , Neoplasias Cutáneas/genética , Animales , Progresión de la Enfermedad , Femenino , Interleucina-10/genética , Interleucina-10/metabolismo , Sistema Calicreína-Quinina/genética , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/secundario , Melanoma Experimental/metabolismo , Melanoma Experimental/secundario , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Proteína Quinasa 1 Activada por Mitógenos/genética , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/genética , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Índice Mitótico , Proteínas Proto-Oncogénicas c-akt/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Receptor de Bradiquinina B1/deficiencia , Transducción de Señal , Neoplasias Cutáneas/metabolismo , Neoplasias Cutáneas/patologíaRESUMEN
The effects of kinin B1 receptor (B1 R) deletion were examined on femur bone regeneration in streptozotocin (STZ)-type 1 diabetes. Diabetes induction in wild-type C57/BL6 (WTC57BL6) mice led to decrease in body weight and hyperglycemia, compared to the non-diabetic group of the same strain. The lack of B1 R did not affect STZ-elicited body weight loss, but partially prevented hyperglycemia. Diabetic mice had a clear delay in bone regeneration, and displayed large areas of loose connective tissue within the defects, with a reduced expression of the mineralization-related protein osteonectin, when compared to the non-diabetic WTC57/BL6. The non-diabetic and diabetic B1 R knockout (B1 RKO) mice had bone regeneration levels and osteonectin expression comparable to that seen in control WTC57/BL6 mice. WTC57/BL6 STZ-diabetic mice also showed a marked reduction of collagen contents, with increased immunolabeling for the apoptosis marker caspase-3, whereas diabetic B1 RKO had collagen levels and caspase-3 activity comparable to those observed in non-diabetic WTC57/BL6 or B1 RKO mice. No significant difference was detected in the number of tartrate-resistant acid phosphatase (TRAP)-stained cells, or in RANK/RANKL/OPG system immunolabeling throughout the experimental groups. Data bring novel evidence on the relevance of kinin B1 R under type 1 diabetes with regards to its role in bone regeneration.
Asunto(s)
Regeneración Ósea , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 1/metabolismo , Fracturas del Fémur/metabolismo , Fémur/metabolismo , Curación de Fractura , Receptor de Bradiquinina B1/deficiencia , Animales , Apoptosis , Caspasa 3/metabolismo , Colágeno/metabolismo , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/patología , Diabetes Mellitus Experimental/fisiopatología , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 1/patología , Diabetes Mellitus Tipo 1/fisiopatología , Fracturas del Fémur/genética , Fracturas del Fémur/patología , Fracturas del Fémur/fisiopatología , Fémur/patología , Fémur/fisiopatología , Masculino , Ratones Endogámicos C57BL , Ratones Noqueados , Osteonectina/metabolismo , Receptor de Bradiquinina B1/genética , Transducción de Señal , Factores de TiempoRESUMEN
Kinins B1 and B2 receptors (B1R and B2R) are classically associated with inflammation, but our group has recently demonstrated new roles for B1R in metabolism using a knockout model (B1 (-/-)). B1 (-/-) mice display improvement on leptin and insulin sensitivity and is protected from high fat diet (HFD)-induced obesity. Here, we evaluate the hepatic effects of the B1R ablation and its role on hepatic function. Despite no expression of hepatic B1R, HFD-induced hepatic lipid accumulation was lower than in control animals. B1 (-/-) mice also presented lower hepatic lipogenesis and SCD1 protein content in the liver. When stimulated with exogenous leptin, B1 (-/-) mice exhibited increased hepatic pJAK2. Similarly, leptin signaling was enhanced in the liver of ob/ob-B1 (-/-) mice, as demonstrated by increased levels of pSTAT3 compared to ob/ob. Plasma concentrations of intercellular adhesion molecule 1, fetuin A, leukemia inhibitory factor, tissue inhibitor of metalloprotease-1, resistin, and oncostatin M were reduced in B1 (-/-). Finally, B1 (-/-) mice have increased gene expression of hepatic B2 receptor, but no difference in leptin receptor expression. Our results show that B1 (-/-) mice are protected from non-alcoholic fatty liver disease (NAFLD) after HFD treatment. Since B1R expression was not observed in the liver after HFD, we propose that the cross talk between the adipose tissue and the liver, mainly through leptin, is an important factor contributing to the observed results. Besides that, several other inflammatory mediators already correlated with NAFLD or liver function were found to be altered in our model. Taken together, our data suggest that B1R plays an important role in hepatic steatosis development.
Asunto(s)
Hígado Graso/metabolismo , Leptina/metabolismo , Hígado/metabolismo , Receptor de Bradiquinina B1/deficiencia , Adipoquinas/sangre , Animales , Dieta Alta en Grasa , Masculino , Ratones , Ratones Noqueados , Enfermedad del Hígado Graso no Alcohólico , Receptor de Bradiquinina B1/genética , Receptor de Bradiquinina B2/metabolismo , Receptores de Leptina/metabolismo , Estearoil-CoA Desaturasa/metabolismoRESUMEN
The Kallikrein-Kinin System (KKS) has been implicated in several aspects of metabolism, including the regulation of glucose homeostasis and adiposity. Kinins and des-Arg-kinins are the major effectors of this system and promote their effects by binding to two different receptors, the kinin B2 and B1 receptors, respectively. To understand the influence of the KKS on the pathophysiology of obesity and type 2 diabetes (T2DM), we generated an animal model deficient for both kinin receptor genes and leptin (obB1B2KO). Six-month-old obB1B2KO mice showed increased blood glucose levels. Isolated islets of the transgenic animals were more responsive to glucose stimulation releasing greater amounts of insulin, mainly in 3-month-old mice, which was corroborated by elevated serum C-peptide concentrations. Furthermore, they presented hepatomegaly, pronounced steatosis, and increased levels of circulating transaminases. This mouse also demonstrated exacerbated gluconeogenesis during the pyruvate challenge test. The hepatic abnormalities were accompanied by changes in the gene expression of factors linked to glucose and lipid metabolisms in the liver. Thus, we conclude that kinin receptors are important for modulation of insulin secretion and for the preservation of normal glucose levels and hepatic functions in obese mice, suggesting a protective role of the KKS regarding complications associated with obesity and T2DM.
Asunto(s)
Glucosa/metabolismo , Homeostasis/fisiología , Hígado/metabolismo , Receptor de Bradiquinina B1/deficiencia , Receptor de Bradiquinina B1/metabolismo , Receptor de Bradiquinina B2/deficiencia , Receptor de Bradiquinina B2/metabolismo , Animales , Glucemia/metabolismo , Composición Corporal/genética , Composición Corporal/fisiología , Homeostasis/genética , Hiperglucemia/sangre , Hiperglucemia/genética , Hiperglucemia/metabolismo , Resistencia a la Insulina/genética , Resistencia a la Insulina/fisiología , Sistema Calicreína-Quinina/genética , Sistema Calicreína-Quinina/fisiología , Ratones , Ratones Noqueados , Ratones Obesos , Fosforilación , Receptor de Bradiquinina B1/genética , Receptor de Bradiquinina B2/genéticaRESUMEN
BACKGROUND AND PURPOSE: In the current study, we investigated the role of both kinin B1 and B2 receptors in peripheral neuropathy induced by the chronic treatment of mice with paclitaxel a widely used chemotherapeutic agent. EXPERIMENTAL APPROACH: Chemotherapy-evoked hyperalgesia was induced by i.p. injections of paclitaxel (2 mg·kg⻹) over 5 consecutive days. Mechanical and thermal hyperalgesia were evaluated between 7 and 21 days after the first paclitaxel treatment. KEY RESULTS: Treatment with paclitaxel increased both mechanical and thermal hyperalgesia in mice (C57BL/6 and CD1 strains). Kinin receptor deficient mice (B1, or B2 receptor knock-out and B1B2 receptor, double knock-out) presented a significant reduction in paclitaxel-induced hypernociceptive responses in comparison to wild-type animals. Treatment of CD1 mice with kinin receptor antagonists (DALBK for B1 or Hoe 140 for B2 receptors) significantly inhibited both mechanical and thermal hyperalgesia when tested at 7 and 14 days after the first paclitaxel injection. DALBK and Hoe 140 were also effective against paclitaxel-induced peripheral neuropathy when given intrathecally or i.c.v. A marked increase in B1 receptor mRNA was observed in the mouse thalamus, parietal and pre-frontal cortex from 7 days after the first paclitaxel treatment. CONCLUSIONS AND IMPLICATIONS: Kinins acting on both B1 and B2 receptors, expressed in spinal and supra-spinal sites, played a crucial role in controlling the hypernociceptive state caused by chronic treatment with paclitaxel.
Asunto(s)
Analgésicos/farmacología , Antagonistas del Receptor de Bradiquinina B1 , Antagonistas del Receptor de Bradiquinina B2 , Bradiquinina/análogos & derivados , Paclitaxel/toxicidad , Enfermedades del Sistema Nervioso Periférico/tratamiento farmacológico , Animales , Bradiquinina/farmacología , Hiperalgesia/inducido químicamente , Hiperalgesia/tratamiento farmacológico , Hiperalgesia/genética , Hiperalgesia/metabolismo , Cininas/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Enfermedades del Sistema Nervioso Periférico/inducido químicamente , Enfermedades del Sistema Nervioso Periférico/genética , Enfermedades del Sistema Nervioso Periférico/metabolismo , ARN Mensajero/genética , Receptor de Bradiquinina B1/deficiencia , Receptor de Bradiquinina B1/genética , Receptor de Bradiquinina B1/metabolismo , Receptor de Bradiquinina B2/deficiencia , Receptor de Bradiquinina B2/genética , Receptor de Bradiquinina B2/metabolismoRESUMEN
BACKGROUND AND PURPOSE: Activation of the proteinase-activated receptor-2 (PAR-2) induces scratching behaviour in mice. Here, we have investigated the role of kinin B(1) and B(2) receptors in the pruritogenic response elicited by activators of PAR-2. EXPERIMENTAL APPROACH: Scratching was induced by an intradermal (i.d.) injection of trypsin or the selective PAR-2 activating peptide SLIGRL-NH(2) at the back of the mouse neck. The animals were observed for 40 min and their scratching response was quantified. KEY RESULTS: I.d. injection of trypsin or SLIGRL-NH(2) evoked a scratching behaviour, dependent on PAR-2 activation. Mice genetically deficient in kinin B(1) or B(2) receptors exhibited reduced scratching behaviour after i.d. injection of trypsin or SLIGRL-NH(2). Treatment (i.p.) with the non-peptide B(1) or B(2)receptor antagonists SSR240612 and FR173657, respectively, prevented the scratching behaviour caused by trypsin or SLIGRL-NH(2). Nonetheless, only treatment i.p. with the peptide B(2)receptor antagonist, Hoe 140, but not the B(1)receptor antagonist (DALBK), inhibited the pruritogenic response to trypsin. Hoe 140 was also effective against SLIGRL-NH(2)-induced scratching behaviour when injected by i.d. or intrathecal (i.t.) routes. Also, the response to SLIGRL-NH(2) was inhibited by i.t. (but not by i.d.) treatment with DALBK. Conversely, neither Hoe 140 nor DALBK were able to inhibit SLIGRL-NH(2)-induced scratching behaviour when given intracerebroventricularly (i.c.v.). CONCLUSIONS AND IMPLICATIONS: The present results demonstrated that kinins acting on both B(1) and B(2) receptors played a crucial role in controlling the pruriceptive signalling triggered by PAR-2 activation in mice.
Asunto(s)
Conducta Animal , Prurito/metabolismo , Receptor de Bradiquinina B1/metabolismo , Receptor de Bradiquinina B2/metabolismo , Receptor PAR-2/metabolismo , Animales , Antipruriginosos/administración & dosificación , Conducta Animal/efectos de los fármacos , Bradiquinina/administración & dosificación , Bradiquinina/análogos & derivados , Antagonistas del Receptor de Bradiquinina B1 , Antagonistas del Receptor de Bradiquinina B2 , Dioxoles/administración & dosificación , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Inyecciones Intradérmicas , Inyecciones Intraperitoneales , Inyecciones Intraventriculares , Inyecciones Espinales , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Oligopéptidos/administración & dosificación , Umbral del Dolor , Prurito/inducido químicamente , Prurito/genética , Prurito/prevención & control , Prurito/psicología , Quinolinas/administración & dosificación , Receptor de Bradiquinina B1/deficiencia , Receptor de Bradiquinina B1/genética , Receptor de Bradiquinina B2/deficiencia , Receptor de Bradiquinina B2/genética , Receptor PAR-2/agonistas , Sulfonamidas/administración & dosificación , Tripsina/administración & dosificaciónRESUMEN
Angiotensin I-converting enzyme (ACE), a common element of renin-angiotensin system (RAS) and kallikrein-kinin system (KKS), is involved in myelopoiesis modulation, mainly by cleaving the tetrapeptide N-acetyl-seryl-aspartyl-lysyl-proline (AcSDKP). Based on this finding and in our results showing B1 and B2 kinin receptors expression in murine bone marrow (BM) cells, we evaluated the ACE influence on myelopoiesis of kinin B1 receptor knockout mice (B1KO) using long-term bone marrow cultures (LTBMCs). Captopril and AcSDKP were used as controls. Enhanced ACE activity, expressed by non-hematopoietic cells (Ter-199(-) and CD45(-)), was observed in B1KO LTBMCs when compared to wild-type (WT) cells. ACE hyperfunction in B1KO cells was maintained when LTBMCs from B1KO mice were treated with captopril (1.0microM) or AcSDKP (1.0nM). Although no alterations were observed in ACE mRNA and protein levels under these culture conditions, 3.0nM of AcSDKP increased ACE mRNA levels in WT LTBMCs. No alteration in the number of GM-CFC was seen in B1KO mice compared to WT animals, even when the former were treated with AcSDKP (10microg/kg) or captopril (100mg/kg) for 4 consecutive days. Hematological data also revealed no differences between WT and B1KO mice under basal conditions. When the animals received 4 doses of lipopolysaccharide (LPS), a decreased number of blood cells was detected in B1KO mice in relation to WT. We also found a decreased percentage of Gr1(+)/Mac-1(+), Ter119(+), B220(+), CD3(+), and Lin(-)Sca1(+)c-Kit(+) (LSK) cells in the BM of B1KO mice compared to WT animals. Low AcSDKP levels were observed in BM cultures from B1KO in comparison to WT cultures. We conclude that ACE hyperfunction in B1KO mice resulted in faster hydrolysis of AcSDKP peptide, which in turn decreased in BM tissues allowing HSC to enter the S stage of the cell cycle.
Asunto(s)
Mielopoyesis/efectos de los fármacos , Oligopéptidos/farmacología , Peptidil-Dipeptidasa A/metabolismo , Receptor de Bradiquinina B1/metabolismo , Animales , Células de la Médula Ósea/citología , Células de la Médula Ósea/metabolismo , Captopril/farmacología , Granulocitos/citología , Granulocitos/efectos de los fármacos , Sistema Calicreína-Quinina , Lipopolisacáridos/toxicidad , Macrófagos/citología , Macrófagos/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Peptidil-Dipeptidasa A/genética , Receptor de Bradiquinina B1/deficiencia , Receptor de Bradiquinina B1/genética , Sistema Renina-AngiotensinaRESUMEN
Previously we have demonstrated that bradykinin B1 receptor deficient mice (B1KO) were protected against renal ischemia and reperfusion injury (IRI). Here, we aimed to analyze the effect of B1 antagonism on renal IRI and to study whether B1R knockout or antagonism could modulate the renal expression of pro and anti-inflammatory molecules. To this end, mice were subjected to 45 minutes ischemia and reperfused at 4, 24, 48 and 120 hours. Wild-type mice were treated intra-peritoneally with antagonists of either B1 (R-954, 200 microg/kg) or B2 receptor (HOE140, 200 microg/kg) 30 minutes prior to ischemia. Blood samples were collected to ascertain serum creatinine level, and kidneys were harvested for gene transcript analyses by real-time PCR. Herein, B1R antagonism (R-954) was able to decrease serum creatinine levels, whereas B2R antagonism had no effect. The protection seen under B1R deletion or antagonism was associated with an increased expression of GATA-3, IL-4 and IL-10 and a decreased T-bet and IL-1beta transcription. Moreover, treatment with R-954 resulted in lower MCP-1, and higher HO-1 expression. Our results demonstrated that bradykinin B1R antagonism is beneficial in renal IRI.
Asunto(s)
Antagonistas del Receptor de Bradiquinina B1 , Enfermedades Renales/tratamiento farmacológico , Daño por Reperfusión/tratamiento farmacológico , Animales , Bradiquinina/análogos & derivados , Bradiquinina/uso terapéutico , Caspasa 3/metabolismo , Muerte Celular , Cartilla de ADN , Factor de Transcripción GATA3/uso terapéutico , Eliminación de Gen , Regulación de la Expresión Génica , Humanos , Interleucina-10/uso terapéutico , Interleucina-4/uso terapéutico , Riñón/fisiología , Riñón/fisiopatología , Enfermedades Renales/mortalidad , Enfermedades Renales/patología , Masculino , Ratones , Ratones Noqueados , Receptor de Bradiquinina B1/deficiencia , Receptor de Bradiquinina B1/genética , Daño por Reperfusión/mortalidad , Daño por Reperfusión/patología , SobrevivientesRESUMEN
OBJECTIVE: Kinins mediate pathophysiological processes related to hypertension, pain, and inflammation through the activation of two G-protein-coupled receptors, named B(1) and B(2). Although these peptides have been related to glucose homeostasis, their effects on energy balance are still unknown. RESEARCH DESIGN AND METHODS: Using genetic and pharmacological strategies to abrogate the kinin B(1) receptor in different animal models of obesity, here we present evidence of a novel role for kinins in the regulation of satiety and adiposity. RESULTS: Kinin B(1) receptor deficiency in mice (B(1)(-/-)) resulted in less fat content, hypoleptinemia, increased leptin sensitivity, and robust protection against high-fat diet-induced weight gain. Under high-fat diet, B(1)(-/-) also exhibited reduced food intake, improved lipid oxidation, and increased energy expenditure. Surprisingly, B(1) receptor deficiency was not able to decrease food intake and adiposity in obese mice lacking leptin (ob/ob-B(1)(-/-)). However, ob/ob-B(1)(-/-) mice were more responsive to the effects of exogenous leptin on body weight and food intake, suggesting that B(1) receptors may be dependent on leptin to display their metabolic roles. Finally, inhibition of weight gain and food intake by B(1) receptor ablation was pharmacologically confirmed by long-term administration of the kinin B(1) receptor antagonist SSR240612 to mice under high-fat diet. CONCLUSIONS: Our data suggest that kinin B(1) receptors participate in the regulation of the energy balance via a mechanism that could involve the modulation of leptin sensitivity.
Asunto(s)
Grasas de la Dieta , Leptina/farmacología , Obesidad/prevención & control , Receptor de Bradiquinina B1/deficiencia , Tejido Adiposo/anatomía & histología , Animales , Composición Corporal , Calorimetría Indirecta , Ratones , Ratones Endogámicos C57BL , Ratones NoqueadosRESUMEN
Increased brain deposition of amyloid beta protein (Abeta) and cognitive deficits are classical signs of Alzheimer's disease (AD) that have been widely associated to inflammatory response. We have recently shown that a single i.c.v. injection of aggregated beta-amyloid peptide-(1-40) (Abeta(1-40)) (400 pmol/mouse) results in marked deficits of learning and memory in mice which are related to oxidative stress and synaptic dysfunction. In the present study, we investigated by means of genetic or pharmacological approaches the role of kinin system in the Abeta(1-40) cognitive effects on the water maze paradigm. Spatial learning and memory deficits observed at 7 days following Abeta(1-40) treatment were significantly reduced by the i.c.v. administration of the selective kinin B(2) receptor antagonist d-Arg-[Hyp(3),Thi(5),D-Tic(7),Oic(8)]-BK (Hoe 140). A similar effect was found in mice lacking kinin B(2) receptor. On the other hand, genetic deletion of the inducible kinin B(1) receptor or its blockage by i.c.v. injection of des-Arg(9)-[Leu(8)]-BK antagonist attenuated only the long-term (30 days after treatment) cognitive deficits induced by Abeta(1-40). Moreover, treatment with Abeta(1-40) resulted in a sustained increase in the expression of the kinin B(1) receptor in the hippocampus and prefrontal cortex of mice, while it did not alter the expression of the kinin B(2) receptor in these brain areas. These findings provide convincing evidence that kinins acting via activation of B(1) and B(2) receptors in the CNS exert a critical role in the spatial learning and memory deficits induced by Abeta peptide in mice. Therefore, selective kinin receptor antagonists, especially the new orally active non-peptide antagonists, might represent drugs of potential interest for the treatment of AD.
Asunto(s)
Antagonistas del Receptor de Bradiquinina B1 , Antagonistas del Receptor de Bradiquinina B2 , Trastornos del Conocimiento/genética , Trastornos del Conocimiento/terapia , Receptor de Bradiquinina B1/deficiencia , Receptor de Bradiquinina B2/deficiencia , Enfermedad de Alzheimer/complicaciones , Péptidos beta-Amiloides , Análisis de Varianza , Animales , Conducta Animal , Bradiquinina/administración & dosificación , Bradiquinina/análogos & derivados , Corteza Cerebral/metabolismo , Trastornos del Conocimiento/etiología , Trastornos del Conocimiento/patología , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Conducta Exploratoria/efectos de los fármacos , Hipocampo/metabolismo , Masculino , Aprendizaje por Laberinto/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Fragmentos de Péptidos , Tiempo de Reacción/efectos de los fármacos , Tiempo de Reacción/genética , Receptor de Bradiquinina B1/genética , Receptor de Bradiquinina B2/genética , Factores de TiempoRESUMEN
The peripheral injection of phorbol myristate acetate (PMA) into the mouse paw induces nociception mediated through activation of protein kinase C (PKC). In the present study, we examine the contribution of kinin B1 receptor to PMA-induced nociception. Nociception was assessed after intraplantar injection of PMA or the B1 receptor agonist des-Arg9-bradykinin in mice. Mechanisms of nociception were studied using the combination of knockout mice, selective drugs, and measurement of B1 receptor mRNA and protein levels. Peripheral injection of PMA (50 pmol/paw) induced a nociceptive behaviour that was abolished by selective B1 receptor antagonist des-Arg9-Leu8-bradykinin or by the B1 receptor gene deletion. Moreover, PMA treatment did not alter B1 receptor mRNA levels, but greatly increased B1 receptor protein levels in the mouse paw. The injection of des-Arg9-bradykinin did not cause nociception in naive mice, but produced marked nociception in animals previously treated with a low dose of PMA (0.5 nmol/paw). The co-treatment of PMA with selective PKC or protein synthesis inhibitors, but not with p38 mitogen activated protein kinase (MAPK) or transcription inhibitors significantly reduced des-Arg9-bradykinin-induced nociception. On the other hand, the co-administration of selective PKC or p38 MAPK inhibitors, but not of protein synthesis or transcription inhibitors, reduced des-Arg9-bradykinin-induced nociception when evaluated in PMA pre-injected animals. These results suggest that the B1 receptor exerts a critical role in the nociception caused by PKC activation in peripheral tissues. Since the PKC pathway is downstream of several pro-inflammatory mediators, B1 receptor stimulation appears to contribute to the acute inflammatory pain process.
Asunto(s)
Dolor/metabolismo , Proteína Quinasa C/metabolismo , Receptor de Bradiquinina B1/fisiología , Animales , Conducta Animal/efectos de los fármacos , Bradiquinina/análogos & derivados , Bradiquinina/uso terapéutico , Carcinógenos , Relación Dosis-Respuesta a Droga , Interacciones Farmacológicas , Activación Enzimática/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/genética , Lipopolisacáridos/farmacología , Masculino , Ratones , Ratones Noqueados , Dolor/inducido químicamente , Dolor/tratamiento farmacológico , Dimensión del Dolor , ARN Mensajero/metabolismo , Receptor de Bradiquinina B1/deficiencia , Acetato de Tetradecanoilforbol/efectos adversos , Factores de TiempoRESUMEN
The kinins have an important role in control of the cardiovascular system. They have been associated with protective effects in the heart tissue. Kinins act through stimulation of two 7-transmembrane G protein-coupled receptors, denoted B(1) and B(2) receptors. However, the physiological relevance of B(1) receptor in the heart has not been clearly established. Using B(1) kinin receptor gene knock-out mice we tested the hypothesis that the B(1) receptor plays an important role in the control of baseline cardiac function. We examined the functional aspects of the intact heart and also in the isolated cardiomyocytes to study intracellular Ca(2+) cycling by using confocal microscopy and whole-cell voltage clamp techniques. We measured heart rate, diastolic and systolic tension, contraction and relaxation rates and, coronary perfusion pressure. Whole-cell voltage clamp was performed to measure L-type Ca(2+) current (I(Ca,L)). The hearts from B(1)(-/-) mice showed smaller systolic tension. The average values for WT and B(1)(-/-) mice were 2.6+/-0.04 g vs. 1.6+/-0.08 g, respectively. This result can be explained, at least in part, by the decrease in the Ca(2+) transient (3.1+/-0.06 vs. 3.4+/-0.09 for B(1)(-/-) and WT, respectively). There was an increase in I(Ca,L) at depolarized membrane potentials. Interestingly, the inactivation kinetics of I(Ca,L) was statistically different between the groups. The coronary perfusion pressure was higher in the hearts from B(1)(-/-) mice indicating an increase in coronary resistance. This result can be explained by the significant reduction of eNOS (NOS-3) expression in the aorta of B(1)(-/-) mice. Collectively, our results demonstrate that B(1) receptor exerts a fundamental role in the mammalian cardiac function.
Asunto(s)
Frecuencia Cardíaca/fisiología , Corazón/fisiología , Contracción Miocárdica/fisiología , Miocitos Cardíacos/metabolismo , Receptor de Bradiquinina B1/metabolismo , Animales , Calcio/metabolismo , Circulación Coronaria/fisiología , Expresión Génica , Silenciador del Gen , Ventrículos Cardíacos/citología , Masculino , Potenciales de la Membrana/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Miocitos Cardíacos/citología , Óxido Nítrico Sintasa de Tipo III/genética , Óxido Nítrico Sintasa de Tipo III/metabolismo , Técnicas de Placa-Clamp , Perfusión , ARN Mensajero/metabolismo , Receptor de Bradiquinina B1/deficiencia , Receptor de Bradiquinina B1/genéticaRESUMEN
A transgenic mouse model, deficient in kinin B(1) receptor (B(1)(-/-)) was used to evaluate the role of B(2) receptor in the smooth muscle stomach fundus. The results showed that the potency of bradykinin (BK) to induce contraction in the gastric tissue was maintained whereas the efficacy was markedly reduced. The angiotensin converting enzyme (ACE) inhibitor captopril potentiated BK-induced effect in wild type (WT) but not in B(1)(-/-) fundus. However, ACE activity detected by the convertion of Ang I to Ang II was inhibited by captopril in both types of gastric tissues. Taking into account the hypothesis that captopril and ACE bind to the B(2) receptor, we suggest that this complex was not formed in the stomach deficient in B(1) receptor. Therefore, our finding strongly support the hypothesis that in smooth muscles that constitutively express the kinin B(1) and B(2) receptors, an interaction between captopril and ACE, B(1) and B(2) receptors should occur forming a complex protein interaction for the potentiating effect of ACE on kinin receptors.
Asunto(s)
Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Captopril/farmacología , Mucosa Gástrica/metabolismo , Contracción Muscular/efectos de los fármacos , Contracción Muscular/genética , Receptor de Bradiquinina B1/genética , Animales , Sinergismo Farmacológico , Ratones , Ratones Noqueados , Contracción Muscular/fisiología , Receptor de Bradiquinina B1/deficiencia , Receptor de Bradiquinina B1/metabolismoRESUMEN
Injury to peripheral nerves often results in a persistent neuropathic pain condition that is characterized by spontaneous pain, allodynia, and hyperalgesia. Nerve injury is accompanied by a local inflammatory reaction in which nerve-associated and immune cells release several pronociceptive mediators. Kinin B1 receptors are rarely expressed in nontraumatized tissues, but they can be expressed after tissue injury. Because B1 receptors mediate chronic inflammatory painful processes, we studied their participation in neuropathic pain using receptor gene-deleted mice. In the absence of neuropathy, we found no difference in the paw-withdrawal responses to thermal or mechanical stimulation between B1 receptor knock-out mice and 129/J wild-type mice. Partial ligation of the sciatic nerve in the wild-type mouse produced a profound and long-lasting decrease in thermal and mechanical thresholds in the paw ipsilateral to nerve lesion. Threshold changed neither in the sham-operated animals nor in the paw contralateral to lesion. Ablation of the gene for the B1 receptor resulted in a significant reduction in early stages of mechanical allodynia and thermal hyperalgesia. Furthermore, systemic treatment with the B1 selective receptor antagonist des-Arg9-[Leu8]-bradykinin reduced the established mechanical allodynia observed 7-28 d after nerve lesion in wild-type mice. Partial sciatic nerve ligation induced an upregulation in B1 receptor mRNA in ipsilateral paw, sciatic nerve, and spinal cord of wild-type mice. Together, kinin B1 receptor activation seems to be essential to neuropathic pain development, suggesting that an oral-selective B1 receptor antagonist might have therapeutic potential in the management of chronic pain.
Asunto(s)
Neuralgia/etiología , Receptor de Bradiquinina B1/deficiencia , Receptor de Bradiquinina B1/fisiología , Neuropatía Ciática/complicaciones , Animales , Bradiquinina/análogos & derivados , Bradiquinina/farmacología , Bradiquinina/uso terapéutico , Antagonistas del Receptor de Bradiquinina B1 , Femenino , Lateralidad Funcional , Expresión Génica/fisiología , Hiperalgesia/genética , Hiperalgesia/fisiopatología , Masculino , Ratones , Ratones Noqueados/fisiología , Neuralgia/terapia , Dimensión del Dolor/métodos , ARN Mensajero/metabolismo , Receptor de Bradiquinina B1/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Piel/metabolismo , Piel/patología , Temperatura Cutánea/genética , Factores de TiempoRESUMEN
The tissue sclerosis found in epilepsy of limbic origin is characterized by shrunken gliotic hippocampus, granule cell loss in the dentate gyrus and extensive pyramidal cell loss in Ammon's horn. Evidence has indicated that sprouting of dentate granule cell axons into the inner molecular layer of the dentate gyrus is related to hyperexcitability. Trying to understand the role of kinin B1 and B2 receptors in the physiopathology of temporal lobe epilepsy (TLE), the present work was delineated to study the development of the epilepsy model induced by pilocarpine in B1 and B2 knockout mice (B1KO and B2KO, respectively). Behavior parameters, cell death and mossy fiber sprouting were analyzed. B1KO mice showed increased latency for the first seizure, associated to a decreased frequency of spontaneous seizures, when compared with wild-type mice. In addition, B1KO mice showed less cell death in all hippocampal formation associated to a reduced grade of mossy fiber sprouting. Furthermore, B2KO mice presented minor duration of the silent period and an increased frequency of spontaneous seizures, when compared with wild-type mice. B2KO and their control lineage showed similar pattern of cell death in the hippocampus, which was very intense when compared with saline-treated animals. The mossy fiber sprouting was also increased in B2KO mice, when compared to wild-type mice and saline-treated animals. Taken together, these data suggest a deleterious effect for kinin B1 receptor and a protective effect for kinin B2 receptor during the development of the temporal lobe epilepsy.
Asunto(s)
Modelos Animales de Enfermedad , Epilepsia/metabolismo , Pilocarpina/toxicidad , Receptor de Bradiquinina B1/fisiología , Receptor de Bradiquinina B2/fisiología , Animales , Epilepsia/inducido químicamente , Epilepsia/patología , Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Hipocampo/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Receptor de Bradiquinina B1/deficiencia , Receptor de Bradiquinina B2/deficienciaRESUMEN
The administration of bradykinin may attenuate ischemia and reperfusion (I/R) injury by acting on B(2)Rs. Blockade of B(2)R has also been shown to ameliorate lesions associated with I/R injury. In an attempt to explain these contradictory results, the objective of the present work was to investigate the role of and interaction between B(1) and B(2) receptors in a model of intestinal I/R injury in mice. The bradykinin B(2)R antagonist (HOE 140) inhibited reperfusion-induced inflammatory tissue injury and delayed lethality. After I/R, there was an increase in the expression of B(1)R mRNA that was prevented by HOE 140. In mice that were deficient in B(1)Rs (B(1)R(-/-) mice), inflammatory tissue injury was abrogated, and lethality was delayed and partially prevented. Pretreatment with HOE 140 reversed the protective anti-inflammatory and antilethality effects provided by the B(1)R(-/-) phenotype. Thus, B(2)Rs are a major driving force for B(1)R activation and consequent induction of inflammatory injury and lethality. In contrast, activation of B(2)Rs may prevent exacerbated tissue injury and lethality, an effect unmasked in B(1)R(-/-) mice and likely dependent on the vasodilatory actions of B(2)Rs. Blockade of B(1)Rs could be a more effective strategy than B(2) or B(1)/B(2) receptor blockade for the treatment of the inflammatory injuries that follow I/R.