Corash L, Chen H Y, Levin J, Baker G, Lu H, Mok Y
Blood. 1987 Jul;70(1):177-85.
We have established a murine model and techniques with which to serially study thrombocytopoiesis after induction of experimental immune thrombocytopenia of variable severity and duration. Bone marrow megakaryocyte ploidy distribution was determined by using unfractionated bone marrow, a polyclonal megakaryocyte-specific probe, and two-color, fluorescence-activated flow cytometry. With these techniques, the modal megakaryocyte ploidy class in normal murine bone marrow was 16N. Serial studies of bone marrow megakaryocyte ploidy after the induction of acute, severe thrombocytopenia (platelet count, less than 0.05 X 10(6) microL) demonstrated no detectable change in the ploidy distribution at 12, 24, and 36 hours after the onset of thrombocytopenia. At 48 hours, the modal ploidy class shifted from 16N to 32N, and the 64N class increased significantly (P less than .001). The ploidy distribution returned to normal 120 hours after the onset of thrombocytopenia. A lesser degree of thrombocytopenia (platelet count reduction to 0.100 to 0.200 X 10(6)/microL) delayed the modal ploidy class shift from 16N to 32N until 72 hours after the onset of thrombocytopenia. Chronic, severe thrombocytopenia (platelet count, less than 0.05 X 10(6)/microL for seven days) resulted in a modal ploidy class shift from 16N to 32N during the thrombocytopenic phase and an enhanced increase in the 64N megakaryocyte class during the recovery phase. Mean platelet volume (MPV) was simultaneously measured on isolated total platelet populations after induction of thrombocytopenia. MPV was significantly increased (P less than .001) as early as eight hours after the onset of acute, severe thrombocytopenia, 40 hours before a shift in the ploidy distribution. Mild thrombocytopenia (platelet count reduction to 0.400 X 10(6)/microL) was not associated with a ploidy shift but did result in a significantly increased MPV (P less than .001). These studies demonstrate that the temporal relationship and magnitude of the effects of thrombocytopenia upon megakaryocyte ploidy distribution are dependent upon the degree and the duration of the thrombocytopenic stimulus and that the effects of experimental thrombocytopenia on platelet volume and megakaryocyte ploidy are dissociated.
我们建立了一种小鼠模型和技术，用于连续研究不同严重程度和持续时间的实验性免疫性血小板减少症诱导后的血小板生成。通过使用未分级的骨髓、多克隆巨核细胞特异性探针和双色荧光激活流式细胞术来确定骨髓巨核细胞倍性分布。利用这些技术，正常小鼠骨髓中巨核细胞倍性类别的众数为16N。对急性、严重血小板减少症（血小板计数低于0.05×10⁶/μL）诱导后的骨髓巨核细胞倍性进行的系列研究表明，血小板减少症发作后12、24和36小时，倍性分布没有可检测到的变化。在48小时时，倍性类别的众数从16N转变为32N，并且64N类别显著增加（P＜0.001）。血小板减少症发作后120小时，倍性分布恢复正常。程度较轻的血小板减少症（血小板计数降至0.100至0.200×10⁶/μL）将倍性类别的众数从16N转变为32N的时间推迟到血小板减少症发作后72小时。慢性、严重血小板减少症（血小板计数低于0.05×10⁶/μL持续7天）导致在血小板减少期倍性类别的众数从16N转变为32N，并在恢复期64N巨核细胞类别增加更为明显。在诱导血小板减少症后，对分离的总血小板群体同时测量平均血小板体积（MPV）。早在急性、严重血小板减少症发作后8小时，即倍性分布转变前40小时，MPV就显著增加（P＜0.001）。轻度血小板减少症（血小板计数降至0.400×10⁶/μL）与倍性转变无关，但确实导致MPV显著增加（P＜0.001）。这些研究表明，血小板减少症对巨核细胞倍性分布影响的时间关系和程度取决于血小板减少刺激的程度和持续时间，并且实验性血小板减少症对血小板体积和巨核细胞倍性的影响是分离的。
