RESUMO
The ß1-adrenergic-receptor (ADRB1) antagonist metoprolol reduces infarct size in acute myocardial infarction (AMI) patients. The prevailing view has been that metoprolol acts mainly on cardiomyocytes. Here, we demonstrate that metoprolol reduces reperfusion injury by targeting the haematopoietic compartment. Metoprolol inhibits neutrophil migration in an ADRB1-dependent manner. Metoprolol acts during early phases of neutrophil recruitment by impairing structural and functional rearrangements needed for productive engagement of circulating platelets, resulting in erratic intravascular dynamics and blunted inflammation. Depletion of neutrophils, ablation of Adrb1 in haematopoietic cells, or blockade of PSGL-1, the receptor involved in neutrophil-platelet interactions, fully abrogated metoprolol's infarct-limiting effects. The association between neutrophil count and microvascular obstruction is abolished in metoprolol-treated AMI patients. Metoprolol inhibits neutrophil-platelet interactions in AMI patients by targeting neutrophils. Identification of the relevant role of ADRB1 in haematopoietic cells during acute injury and the protective role upon its modulation offers potential for developing new therapeutic strategies.
Assuntos
Antagonistas de Receptores Adrenérgicos beta 1/farmacologia , Antagonistas de Receptores Adrenérgicos beta 1/uso terapêutico , Metoprolol/farmacologia , Metoprolol/uso terapêutico , Infarto do Miocárdio/tratamento farmacológico , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Neutrófilos/efeitos dos fármacos , Animais , Movimento Celular/efeitos dos fármacos , Células-Tronco Hematopoéticas/metabolismo , Humanos , Metoprolol/administração & dosagem , Camundongos , Infarto do Miocárdio/patologia , Traumatismo por Reperfusão Miocárdica/patologia , Neutrófilos/citologia , Agregação Plaquetária/efeitos dos fármacos , RNA Mensageiro/genética , Receptores Adrenérgicos beta 1/genética , Receptores Adrenérgicos beta 1/metabolismoRESUMO
Hematopoietic stem cell transplantation or bone marrow transplantation is a common approach to reconstitute the immune system of mice that have been subjected to marrow-ablative doses of radiation. This method can be used in the field of atherosclerosis to assess the contribution of hematopoietic cells of a desired genotype to disease pathogenesis. The engraftment of atherosclerosis-prone mice with donor cells that contain genetic alterations in cells of the innate or adaptive immune system has been invaluable to define the role of multiple gene products in atherosclerosis. Here, we describe the different steps involved in the bone marrow transplantation protocol along with specific guidelines regarding the theoretical and technical details of the procedure.
Assuntos
Aterosclerose/cirurgia , Transplante de Medula Óssea/métodos , Transplante de Células-Tronco Hematopoéticas/métodos , Imunidade Adaptativa , Animais , Aterosclerose/genética , Aterosclerose/imunologia , Aterosclerose/metabolismo , Modelos Animais de Doenças , Feminino , Imunidade Inata , Masculino , Camundongos , Quimeras de Transplante , Irradiação Corporal TotalRESUMO
Beyond its well-established roles in mediating leukocyte rolling, E-selectin is emerging as a multifunctional receptor capable of inducing integrin activation in neutrophils, and of regulating various biological processes in hematopoietic precursors. Although these effects suggest important homeostatic contributions of this selectin in the immune and hematologic systems, the ligands responsible for transducing these effects in different leukocyte lineages are not well defined. We have characterized mice deficient in E-selectin ligand-1 (ESL-1), or in both P-selectin glycoprotein-1 (PSGL-1) and ESL-1, to explore and compare the contributions of these glycoproteins in immune and hematopoietic cell trafficking. In the steady state, ESL-1 deficiency resulted in a moderate myeloid expansion that became more prominent when both glycoproteins were eliminated. During inflammation, PSGL-1 dominated E-selectin binding, rolling, integrin activation, and extravasation of mature neutrophils, but only the combined deficiency in PSGL-1 and ESL-1 completely abrogated leukocyte recruitment. Surprisingly, we find that the levels of ESL-1 were strongly elevated in hematopoietic progenitor cells. These elevations correlated with a prominent function of ESL-1 for E-selectin binding and for migration of hematopoietic progenitor cells into the bone marrow. Our results uncover dominant roles for ESL-1 in the immature compartment, and a functional shift toward PSGL-1 dependence in mature neutrophils.