环磷酰胺诱发小鼠血小板减少症模型的建立及其血小板功能的测定

Establishment of a mouse model of cyclophosphamide-induced thrombocytopenia and determination of platelet function in this model

目的建立环磷酰胺诱发急性造血功能抑制的动物模型,探讨血小板在骨髓抑制中发生变化的基础和意义。方法采用环磷酰胺首次大剂量200 mg/kg·b.w.尾静脉注射腺磷酸,次日连续7 d以维持剂量30 mg/kg·b.w.腹腔注射。观察其外周血和骨髓液中细胞数量和形态的变化,用光密度法测定ADP诱导的血小板聚集百分率;玻璃毛细管法测定凝血时间,评价血小板功能的改变。结果用该方法诱导小鼠后,其红细胞、白细胞、血小板和骨髓有核细胞计数均明显减少(P<0.01),血象和骨髓象检查显示正常造血组织被抑制,其中血小板的降低最为显著,达到49%。血小板平均血小板体积和血小板聚集百分率未有改变(P>0.01),凝血时间缩短。结论环磷酰胺可以快速、高效诱导小鼠急性造血功能障碍,尤其血小板减少模型发生时间快,反应强度强,而对血小板功能未有影响,其模型结果稳定,发生率较高。

Objective To establish a mouse model of acute hematopoietic failure and explore the pathological basis ofplatelet changes in bone marrow suppression. Methods An initial large dose of cyclophosphamide (200 mg/kg) wasinjected through the tail veins of the mice, follow by daily intraperitoneal injection starting on the next day for 7 d. Thenumber and morphology of the cells in the peripheral blood and bone marrow were observed by means of cell counting andsmear, respectively, with the platelet aggregation determined using ADP. The coagulation time was measure by turbidimetry.Results The amount of platelets, erythrocytes, leukocytes and the nucleated cells in mice bone marrow was significantlylowered in the mouse models in comparison with the control mice, and the normal hematopoietic tissues were depressed.The blood platelets in the models were lowered by 49%, an amplitude of declination significantly greater than that occurring inthe erythrocytes and leukocytes (by 28% and 25% respectively). Though the mean platelet volume and percent plateletaggregation underwent no obvious changes, the coagulation time was significantly shortened. Conclusion The methods wedescribedforestablishingmousemodelsofhematopoieticfailureinducedbycyclophosphamideisrapidandhighlyefficient, and may facilitate ready preparation of mouse models of thrombocytopenia which respond strongly to cyclopho-sphamide without blood platelets function impairment.