抗血小板GPⅠbα抗体诱导建立原发免疫性血小板减少症模型的研究

A Study on the establishment of immune thrombocytopenia model induced by anti-platelet GP I bα antibodies

目的探讨应用抗血小板膜糖蛋白GPⅠbα抗体AN51、R300建立原发免疫性血小板减少症（ITP）动物模型。方法将20只6-8周豚鼠和裸鼠分别分为对照组和3个实验组，每组5只。豚鼠对照组和实验组分别静脉注射0.2 μg/g IgG和0.05、0.1、0.2 μg/g AN51，裸鼠对照组和实验组分别腹腔注射0.2 μg/g IgG和0.05、0.1、0.2 μg/g R300。在注射后不同时间点对豚鼠和裸鼠行眼眶后静脉丛采血，应用血细胞分析仪检测血小板水平。结果①静脉注射AN51后5 min，0.05、0.1、0.2 μg/g AN51组豚鼠血小板计数分别减低0-5%、50%-60%、70%-80%，0.2 μg/g组下降最为明显，与对照组比较差异有统计学意义（P〈0.001）。②腹腔注射R300后6 h，0.05、0.1、0.2 μg/g R300组裸鼠血小板计数分别下降20%-30%、60%-70%、80%-90%，0.2 μg/g组下降最为明显，与对照组比较差异有统计学意义（P〈0.001）。每天1次持续注射0.2 μg/g R300 1-2周的裸鼠出现典型ITP出血症状（全身大量瘀点、瘀斑，多见于四肢、头部和腹部）。结论0.2 μg/g AN51静脉注射、0.2 μg/g R300腹腔注射可以分别建立豚鼠、裸鼠ITP模型。

ObjectiveTo establish primary immune thrombocytopenia （ITP） animal model induced by anti-platelet membrane glycoprotein GPⅠbα antibodies AN51 and R300.MethodsTwenty 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.Results①Five 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 （P〈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 （P〈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.ConclusionAN51 at 0.2 μg/g and R300 at 0.2 μg/g could establish stable ITP model in guinea pigs and nude mice respectively.