RESUMEN
Expression of mRNA for adenosine receptor subtypes A(1), A(2a), A(2b), and A(3) in normal and lipopolysaccharide (LPS)-activated murine RAW 264.7 macrophages has been investigated using the method of quantitative real-time polymerase chain reaction. The results have shown a very low, unquantifiable expression of adenosine A(1) receptor mRNA in both normal and LPS-activated macrophages. The other three adenosine receptor mRNAs have been found to be expressed at various but always quantifiable levels. Activation of the macrophages by LPS induced upregulation of the expression of adenosine receptor A(2a) and A(2b) mRNA, whereas the expression of adenosine receptor A(3) mRNA was downregulated. Unstimulated macrophages exhibited a high expression of the A(2b) adenosine receptor mRNA. The findings are discussed from the point of view of the antiinflammatory and hematopoiesis-stimulating roles of the adenosine receptor signaling.
Asunto(s)
Hematopoyesis , Lipopolisacáridos/farmacología , Activación de Macrófagos , Macrófagos/metabolismo , ARN Mensajero/metabolismo , Receptores Purinérgicos P1/genética , Animales , Línea Celular , Regulación de la Expresión Génica , Hematopoyesis/efectos de los fármacos , Hematopoyesis/genética , Activación de Macrófagos/efectos de los fármacos , Activación de Macrófagos/genética , Macrófagos/efectos de los fármacos , Ratones , Reacción en Cadena de la Polimerasa , Receptor de Adenosina A1/genética , Receptor de Adenosina A2A/genética , Receptor de Adenosina A2B/genética , Receptor de Adenosina A3/genética , Receptores Purinérgicos P1/efectos de los fármacos , Factores de TiempoRESUMEN
Four mouse bone marrow or thymus cell populations, namely granulopoietic/monocytopoietic, erythropoietic, B-lymphopoietic, and T-lymphopoietic precursor cells have been assayed by RT-PCR technique for the presence and relative amounts of adenosine A(1), A(2a), A(2b), and A(3) receptor mRNA. It has been found that (i) all four populations studied express all four adenosine receptor subtypes, (ii) the A(1), receptor is the least expressed in all populations studied, (iii) the A(3) receptor is markedly expressed in the populations of granulopoietic/monocytopoietic and erythropoietic cells, (iv) the A(2a) receptor is markedly expressed in the populations of B-lymphopoietic and T-lymphopoietic cells, and v) the A(2b) receptor does not predominate in any of the precursor cells studied. Our data offer a new possibility for the assessment of the readiness of these cells to respond, by receptor-mediated mechanisms, to adenosine or its analogs present in the tissues as a result of endogenous processes and/or following their administration.
Asunto(s)
Hematopoyesis , Células Madre Hematopoyéticas/metabolismo , ARN Mensajero/metabolismo , Receptores Purinérgicos P1/genética , Animales , Separación Celular , Femenino , Regulación de la Expresión Génica , Hematopoyesis/genética , Ratones , Ratones Endogámicos C57BL , Reacción en Cadena de la Polimerasa , Receptor de Adenosina A1/genética , Receptor de Adenosina A2A/genética , Receptor de Adenosina A2B/genética , Receptor de Adenosina A3/genéticaRESUMEN
Adenosine A(3) receptor agonist N(6)-(3-iodobenzyl)adenosine-5'-N-methyluronamide (IB-MECA) has been tested from the point of view of potentiating the effects of hematopoietic growth factors interleukin-3 (IL-3), stem cell factor (SCF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and granulocyte colony-stimulating factor (G-CSF) on the growth of hematopoietic progenitor cells for granulocytes and macrophages (GM-CFC) in suspension of normal mouse bone marrow cells in vitro. IB-MECA alone induced no GM-CFC growth. Significant elevation of numbers of GM-CFC evoked by the combinations of IB-MECA with IL-3, SCF, or GM-CSF as compared with these growth factors alone has been noted. Combination of IB-MECA with G-CSF did not induce significantly higher numbers of GM-CFC in comparison with G-CSF alone. Joint action of three drugs, namely of IB-MECA + IL-3 + GM-CSF, produced significantly higher numbers of GM-CFC in comparison with the combinations of IB-MECA + IL-3, IB-MECA + GM-CSF, or IL-3 + GM-CSF. These results give evidence of a significant role of selective activation of adenosine A(3) receptors in stimulation of the growth of granulocyte/ macrophage hematopoietic progenitor cells.
Asunto(s)
Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Células Madre Hematopoyéticas/efectos de los fármacos , Células Madre Hematopoyéticas/metabolismo , Factor de Crecimiento de Hepatocito/farmacología , Interleucina-3/farmacología , Receptor de Adenosina A3/metabolismo , Adenosina/análogos & derivados , Adenosina/farmacología , Animales , División Celular/efectos de los fármacos , Células Cultivadas , Sinergismo Farmacológico , Granulocitos/citología , Células Madre Hematopoyéticas/citología , Técnicas In Vitro , Macrófagos/citología , Masculino , Ratones , Ratones EndogámicosRESUMEN
Meloxicam, a selective inhibitor of cyclooxygenase 2, was tested to determine its ability to modulate hematopoiesis and to influence survival of mid-lethally gamma-irradiated mice. A single dose of meloxicam (20 mg/kg) administered to mice intraperitoneally 1 h before irradiation was shown to enhance serum levels of granulocyte colony-stimulating factor (G-CSF) during the first 24 h after irradiation, to elevate numbers of granulocytic precursor cells in bone marrow and granulocyte counts in peripheral blood on day 10 after irradiation, and to increase 30-day survival of these mice. The results provide new evidence for the protective ability of meloxicam administration to mice irradiated with mid-lethal doses and contribute to the understanding of the mechanisms of this meloxicam action by drawing attention to the possible role of increased endogenous G-CSF production.
Asunto(s)
Inhibidores de la Ciclooxigenasa 2/farmacología , Factor Estimulante de Colonias de Granulocitos/biosíntesis , Protectores contra Radiación/farmacología , Tiazinas/farmacología , Tiazoles/farmacología , Animales , Hematopoyesis/efectos de la radiación , Masculino , Meloxicam , RatonesRESUMEN
Hematopoiesis-modulating action of meloxicam, a cyclooxyge-nase-2 inhibitor, has been evaluated in mice. Increased serum level of granulocyte colony-stimulating factor (G-CSF) after meloxicam administration has been found in sublethally gamma-irradiated animals. In further experiments hematopoiesis-stimulating effects of meloxicam and G-CSF given alone or in combination have been investigated. Granulocyte/macrophage progenitor cells counts were used to monitor these effects. Meloxicam and exogenous G-CSF did not act synergistically when given in combination, but could be mutually substituted during their repeated administration. The results suggest a promising possibility of using meloxicam as an auxiliary drug reducing the high costs of G-CSF therapy of myelosuppression.
Asunto(s)
Médula Ósea/efectos de los fármacos , Inhibidores de la Ciclooxigenasa 2/farmacología , Factor Estimulante de Colonias de Granulocitos/sangre , Hematopoyesis/efectos de los fármacos , Tiazinas/farmacología , Tiazoles/farmacología , Animales , Médula Ósea/efectos de la radiación , Diferenciación Celular , Interacciones Farmacológicas , Quimioterapia Combinada , Factor Estimulante de Colonias de Granulocitos/administración & dosificación , Factor Estimulante de Colonias de Granulocitos/efectos de los fármacos , Granulocitos/citología , Granulocitos/efectos de los fármacos , Granulocitos/efectos de la radiación , Leucopenia/prevención & control , Masculino , Meloxicam , Ratones , Ratones Endogámicos CBA , Irradiación Corporal TotalRESUMEN
Our previous studies have shown that the combined administration of drugs elevating extracellular adenosine, i.e. dipyridamole (DP) and adenosine monophosphate (AMP), enhances murine hematopoiesis and potentiates the action of granulocyte colony-stimulating factor (G-CSF). In this study, colony-stimulating activity (CSA) of blood serum of mice treated with DP+AMP, G-CSF or all these drugs in combination, i.e. the ability of the sera to stimulate the growth of GM-CFC colonies, was assayed in vitro. Furthermore, the concentration of GM-CSF and IL-6 in the sera was determined. Administration of DP+AMP was found to enhance significantly serum CSA at all time intervals of serum sampling including 24 h after the last injection of the tested drugs. Additive effects of DP+AMP and G-CSF on serum CSA were noted at early intervals after administration of the drugs. Furthermore, IL-6 levels were significantly elevated in the sera of mice which were administered DP+AMP either alone or in combination with G-CSF. Our results show that the effects of DP+AMP are indirect, mediated through the induction of some cytokine(s) and/or growth factor(s) and that extracellular adenosine can act in cooperation with G-CSF. These findings contribute to the further elucidation of the role of adenosine in hematopoiesis.
Asunto(s)
Adenosina Monofosfato/metabolismo , Adenosina/metabolismo , Dipiridamol/farmacología , Factor Estimulante de Colonias de Granulocitos/metabolismo , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Hematopoyesis/efectos de los fármacos , Células Madre Hematopoyéticas/efectos de los fármacos , Interleucina-6/metabolismo , Adenosina Monofosfato/farmacología , Animales , Ensayo de Unidades Formadoras de Colonias , Femenino , Factor Estimulante de Colonias de Granulocitos/farmacología , Factor Estimulante de Colonias de Granulocitos y Macrófagos/sangre , Células Madre Hematopoyéticas/metabolismo , Interleucina-6/sangre , Masculino , Ratones , Ratones Endogámicos ICR , Factores de Tiempo , Regulación hacia ArribaRESUMEN
Meloxicam, a selective inhibitor of cyclooxygenase 2, a nonsteroidal anti-inflammatory drug with an improved side-effects profile in terms of gastrointestinal toxicity, has been found to stimulate hematopoiesis in whole-body gamma-irradiated mice. A distinct corroboration of this positive action of meloxicam is an enhancement of the recovery of hematopoietic progenitor cells committed to granulocyte-macrophage and erythroid development, which has been demonstrated in sublethally irradiated animals treated with meloxicam at a dose of 20 mg/kg administered intraperitoneally either singly 1 h before irradiation or repeatedly after radiation exposure. The results suggest that meloxicam can be added to the list of biological response modifiers that can be used in the treatment of hematopoietic damage induced by ionizing radiation.
Asunto(s)
Médula Ósea/efectos de los fármacos , Médula Ósea/efectos de la radiación , Inhibidores de la Ciclooxigenasa 2/administración & dosificación , Rayos gamma/efectos adversos , Hematopoyesis/efectos de los fármacos , Hematopoyesis/efectos de la radiación , Protectores contra Radiación/administración & dosificación , Tiazinas/administración & dosificación , Tiazoles/administración & dosificación , Irradiación Corporal Total/efectos adversos , Animales , Médula Ósea/lesiones , Médula Ósea/patología , Células Cultivadas , Masculino , Meloxicam , Ratones , Dosis de RadiaciónRESUMEN
A single dose of IMUNOR, a low-molecular-weight immunodulator prepared from disintegrated and ultrafiltered pig leukocytes, was found to enhance recovery of murine pool of hemopoietic progenitor cells for granulocytes and macrophages damaged by a single injection of cytotoxic drugs 5-fluorouracil or cisplatin. The best results were obtained after the treatment with IMUNOR on days 3 or 4 after the injection of 5-fluorouracil or cisplatin. These results together with previous findings obtained in our laboratory suggest that IMUNOR has the potential to become a part of treatment schemes in oncological practice aimed at alleviation of myelosuppression evoked by cytotoxic anti-tumor therapy.
Asunto(s)
Antimetabolitos Antineoplásicos , Antineoplásicos , Enfermedades de la Médula Ósea/inducido químicamente , Enfermedades de la Médula Ósea/tratamiento farmacológico , Cisplatino , Fluorouracilo , Factores Inmunológicos/farmacología , Leucocitos/química , Leucocitos/inmunología , Animales , Células de la Médula Ósea/efectos de los fármacos , Células de la Médula Ósea/inmunología , Femenino , Granulocitos/efectos de los fármacos , Granulocitos/inmunología , Hematopoyesis/efectos de los fármacos , Factores Inmunológicos/química , Ratones , Ratones Endogámicos ICR , Células Madre/efectos de los fármacos , Porcinos , UltrafiltraciónRESUMEN
The aim of the studies was to ascertain if adenosine is able to co-operate with selected hematopoietic growth factors and cytokines, namely with granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), stem cell factor (SCF), interleukin-3 (IL-3), and interleukin-11 (IL-11), in inducing the growth of colonies from hematopoietic progenitor cells for granulocytes and macrophages (GM-CFC) from normal bone marrow cells in vitro. Adenosine was found not to produce any colonies when present in the cultures as the only potential stimulator. All the tested cytokines and growth factors were observed to induce the growth of distinct numbers of GM-CFC colonies, with the exception of IL-11. When suboptimal concentrations of the evaluated cytokines and growth factors were tested in the cultures in which various concentrations of adenosine were concomitantly present, mutually potentiating effects were found in the case of IL-3 and SCF. These results confirm the role of adenosine in regulation of granulopoiesis and predict IL-3 and SCF as candidates for further in vivo studies of their combined administration with adenosine.