Fetal liver tissues obtained from 28 human fetuses with gestation age from 3 to 6 months and fetal bone marrow from 35 human fetuses from 3 to 7 months were observed by immunochemical staining with anti-platelet GPⅡ ...Fetal liver tissues obtained from 28 human fetuses with gestation age from 3 to 6 months and fetal bone marrow from 35 human fetuses from 3 to 7 months were observed by immunochemical staining with anti-platelet GPⅡ b / Ⅲa monoclonal antibody and ABC technique. In the fetal liver, megakaryocytes were wholly located among growing fetal liver cells and near foci of hemopoiesis. Some megakaryocytes in the fetal liver were small7890- lymphoid-like megakaryocytes. The size of megakaryocytes both in the fetal liver (14.79 ± 4.52μm) and in the fetal bone marrow (16.08±7.39 μm) was small, which did not vary significantly over the gestation age ranging from 3 to 6 or 7 months. However, the maturation stage of megakaryocytes in the fetal liver shifted to more mature stage with the advancement of gestation, although the maturation stage of megakaryocytes in the fetal bone marrow did not change with the advancement of gestation from 4 to 7 months, the megakaryocyte in the fetal bone marrow was less mature展开更多
OBJECTIVE Salvia miltiorrhiza bunge contains various active constituents,some of which have been developed as commercially available medicine.Moreover,some other ingredients in Salvia miltiorrhiza play great roles in ...OBJECTIVE Salvia miltiorrhiza bunge contains various active constituents,some of which have been developed as commercially available medicine.Moreover,some other ingredients in Salvia miltiorrhiza play great roles in anti-platelet activity.The aim of the present study was to investigate the effects and the underlying mechanism of miltirone,a lipophilic compound of Salvia miltiorrhiza Bunge.METHODS The ability of miltirone to modulate platelet function was investigated by a variety of in vitro and in vivo experiments.Platelet aggregation and dense granule secretion induced by various agonists were measured with platelet aggregometer.Clot retraction and spreading were imaged by digital camera and fl uorescence microscope.Ferric chloride-induced carotid injury model and pulmonary thromboembolism model were used to check miltirone effect in vivo.To elucidate the mechanisms of anti-platelet activity of miltirone,flow cytometry and Western blotting were performed.RESULTS Miltirone(2,4,8 μmol·L^(-1)) was shown to suppress platelet aggregation,dense granule and α granule secretion in a dose-dependent manner.Meanwhile,miltirone inhibited the clot retraction and spreading of washed platelets.It reduced the phosphorylation of PLCγ2,PKC,Akt,GSK3β and ERK1/2 in the downstream signal pathway of collagen receptor.It also reduced the phosphorylation of Src and FAK in the integrin αⅡbβ3 mediated "outside-in" signaling,while it did not suppress the phosphorylation of β3.In addition,miltirone prolonged the occlusion time and reduced collagen/epinephrine induced pulmonary thrombi.CONCLUSION Miltirone suppresses platelet "inside-out" and "outside-in" signaling by affecting PLCγ2/PKC/ERK1/2,PI3K/Akt and Src/FAK signaling.Therefore,miltirone might represent a potential anti-platelet candidate for the prevention of thrombotic disorders.展开更多
文摘Fetal liver tissues obtained from 28 human fetuses with gestation age from 3 to 6 months and fetal bone marrow from 35 human fetuses from 3 to 7 months were observed by immunochemical staining with anti-platelet GPⅡ b / Ⅲa monoclonal antibody and ABC technique. In the fetal liver, megakaryocytes were wholly located among growing fetal liver cells and near foci of hemopoiesis. Some megakaryocytes in the fetal liver were small7890- lymphoid-like megakaryocytes. The size of megakaryocytes both in the fetal liver (14.79 ± 4.52μm) and in the fetal bone marrow (16.08±7.39 μm) was small, which did not vary significantly over the gestation age ranging from 3 to 6 or 7 months. However, the maturation stage of megakaryocytes in the fetal liver shifted to more mature stage with the advancement of gestation, although the maturation stage of megakaryocytes in the fetal bone marrow did not change with the advancement of gestation from 4 to 7 months, the megakaryocyte in the fetal bone marrow was less mature
基金supported by National Natural Science Fund of China(81273574) Chinese Herb Key Project by Health and Family Planning Commission of Hubei Province
文摘OBJECTIVE Salvia miltiorrhiza bunge contains various active constituents,some of which have been developed as commercially available medicine.Moreover,some other ingredients in Salvia miltiorrhiza play great roles in anti-platelet activity.The aim of the present study was to investigate the effects and the underlying mechanism of miltirone,a lipophilic compound of Salvia miltiorrhiza Bunge.METHODS The ability of miltirone to modulate platelet function was investigated by a variety of in vitro and in vivo experiments.Platelet aggregation and dense granule secretion induced by various agonists were measured with platelet aggregometer.Clot retraction and spreading were imaged by digital camera and fl uorescence microscope.Ferric chloride-induced carotid injury model and pulmonary thromboembolism model were used to check miltirone effect in vivo.To elucidate the mechanisms of anti-platelet activity of miltirone,flow cytometry and Western blotting were performed.RESULTS Miltirone(2,4,8 μmol·L^(-1)) was shown to suppress platelet aggregation,dense granule and α granule secretion in a dose-dependent manner.Meanwhile,miltirone inhibited the clot retraction and spreading of washed platelets.It reduced the phosphorylation of PLCγ2,PKC,Akt,GSK3β and ERK1/2 in the downstream signal pathway of collagen receptor.It also reduced the phosphorylation of Src and FAK in the integrin αⅡbβ3 mediated "outside-in" signaling,while it did not suppress the phosphorylation of β3.In addition,miltirone prolonged the occlusion time and reduced collagen/epinephrine induced pulmonary thrombi.CONCLUSION Miltirone suppresses platelet "inside-out" and "outside-in" signaling by affecting PLCγ2/PKC/ERK1/2,PI3K/Akt and Src/FAK signaling.Therefore,miltirone might represent a potential anti-platelet candidate for the prevention of thrombotic disorders.
