Zhou K X, Yan R, Chen M X, Liu J, Cui Q Y, Hu R P, Liu Y C, Zhang Y, Ruan C G, Dai K S
Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University; Key Laboratory of Thrombosis and Hemostasis, Ministry of Health; Collaborative Innovation Center of Hematology, Soochow University, Suzhou 215006, China.
Zhonghua Xue Ye Xue Za Zhi. 2017 May 14;38(5):390-393. doi: 10.3760/cma.j.issn.0253-2727.2017.05.007.
To establish primary immune thrombocytopenia (ITP) animal model induced by anti-platelet membrane glycoprotein GPⅠbα antibodies AN51 and R300. Twenty guinea pigs (6-8 week) were divided into 4 groups. Five guinea pigs in each group were intravenously injected with different doses of AN51 (0.05, 0.1, 0.2 μg/g) and 0.2 μg/g IgG as control. The whole blood was collected from inner angular venous plexus. Platelets number was determined by an automated cell counter and Swiss-Jim method. Then, the similar protocol was used to establish ITP nude mice model by intraperitoneal injection of different concentrations of anti-platelet GPⅠbα antibody R300, respectively. ①Five minutes after intravenous injection of AN51 at 0.05, 0.1 and 0.2 μg/g, the platelet counts of guinea pigs reduced about 0-5%, 50%-60% and 70%-80% compared to the control group, respectively. The difference was statistically significant (<0.01) . ②Six hours after intraperitoneal injection of R300 at 0.05, 0.1, 0.2 μg/g, the platelet counts of nude mice decreased about 20%-30%, 60%-70% and 80%-90% compared to the control group, respectively. The difference was statistically significant (<0.01) . The nude mice, injected 0.2 μg/g R300 once a day for 2 weeks, showed typical ITP clinical manifestations including large number of petechiaes or ecchymoses on limbs, head and abdomen. AN51 at 0.2 μg/g and R300 at 0.2 μg/g could establish stable ITP model in guinea pigs and nude mice respectively.
建立抗血小板膜糖蛋白GPⅠbα抗体AN51和R300诱导的原发性免疫性血小板减少症(ITP)动物模型。将20只豚鼠(6 - 8周龄)分为4组。每组5只豚鼠分别静脉注射不同剂量的AN51(0.05、0.1、0.2μg/g)以及作为对照的0.2μg/g IgG。从内眦静脉丛采集全血。通过自动血细胞计数器和瑞氏-吉姆萨法测定血小板数量。然后,采用类似方案分别通过腹腔注射不同浓度的抗血小板GPⅠbα抗体R300建立ITP裸鼠模型。①静脉注射0.05、0.1和0.2μg/g的AN51后5分钟,豚鼠的血小板计数与对照组相比分别降低约0 - 5%、50% - 60%和70% - 80%。差异具有统计学意义(<0.01)。②腹腔注射0.05、0.1、0.2μg/g的R300后6小时,裸鼠的血小板计数与对照组相比分别降低约20% - 30%、60% - 70%和80% - 90%。差异具有统计学意义(<0.01)。每天一次腹腔注射0.2μg/g R300,持续2周的裸鼠表现出典型的ITP临床表现,包括四肢、头部和腹部出现大量瘀点或瘀斑。0.2μg/g的AN51和0.2μg/g的R300可分别在豚鼠和裸鼠中建立稳定的ITP模型。