血小板聚集在新生犬动脉导管闭合过程中的作用  

Role of platelets aggregation in the process of ductus arteriosus closure of newborn pups

作　　者：任漪 高翔羽[1] 赵丹丹 王乐瑶 黄迪 苏敏[1] Ren Yi;Gao Xiangyu;Zhao Dandan;Wang Leyao;Huang Di;Su Min(Department of Neonatology,Xuzhou Central Hospital(the Xuzhou Clinical College of Xuzhou Medical University),Xuzhou 221009,Jiangsu,China)

机构地区：[1]徐州市中心医院(徐州医科大学徐州临床学院)新生儿科,江苏徐州221009

出　　处：《中华危重病急救医学》2022年第11期1194-1199,共6页Chinese Critical Care Medicine

基　　金：江苏省青年医学重点人才培养项目(QNRC2016385);江苏省徐州市科技局应用基础研究计划项目(KC18034)。

摘　　要：目的探讨血小板聚集在新生犬动脉导管闭合发生发展过程中的作用,以及血小板膜糖蛋白Ⅱb-Ⅲa(GPⅡb-Ⅲa)受体拮抗剂替罗非班的影响。方法选取24月龄比格母犬4只,分别编号为1、2、3、4,分两批在预产期前1~2 d进行剖宫产取出仔犬。母犬1和2为第一批,经剖宫产取出新生犬18只,作为对照组,按出生后时间点分为1、4、12 h 3个亚组,每个亚组6只;新生犬出生后即刻经颈静脉注射生理盐水10 mL/kg。母犬3和4为第二批,经剖宫产取出新生犬19只,作为替罗非班组,按出生后时间点分为1 h(n=6)、4 h(n=6)、12 h(n=7)3个亚组;新生犬出生后即刻经颈静脉注射盐酸替罗非班注射液10 mL/kg(10 mL注射液含替罗非班2.5 mg)。采用超声心动图测量动脉导管内径;手术剥离取出动脉导管并分成两部分,分别采用蛋白质免疫印迹试验(Western blotting)和免疫组化法检测血小板膜GPⅡb-Ⅲa表达。结果对照组1 h亚组中有1只新生犬于出生后0.5 h死亡;替罗非班组19只均顺利完成实验。出生后1 h,对照组和替罗非班组新生犬动脉导管均未闭合,导管内径差异无统计学意义(mm:1.72±0.08比1.70±0.11,P>0.05);出生后4 h,对照组有1只新生犬动脉导管闭合,替罗非班组均未闭合,且动脉导管内径明显大于对照组(mm:1.52±0.15比0.95±0.48,P<0.05);出生后12 h,对照组新生犬动脉导管均闭合,替罗非班组仍有2只未闭合,导管内径分别为1.0 mm和1.1 mm。Western blotting结果显示,两组新生犬出生后1、4、12 h动脉导管内血小板膜GPⅡb-Ⅲa蛋白表达均呈逐渐升高趋势;替罗非班组1 h GPⅡb-Ⅲa蛋白表达明显低于对照组(GPⅡb-Ⅲa/β-actin:0.67±0.07比0.84±0.16,P<0.05),4 h和12 h时GPⅡb-Ⅲa蛋白表达略低于对照组(GPⅡb-Ⅲa/β-actin:4 h为0.85±0.12比0.95±0.11,12 h为1.04±0.16比1.09±0.17,均P>0.05)。免疫组化结果显示,两组新生犬动脉导管内血小板膜GPⅡb-Ⅲa表达的变化趋势与Western bloObjective To investigate the role of platelets aggregation in the developing process of ductus arteriosus closure of newborn pups,and the effect of platelet membrane glycoproteinⅡb-Ⅲa(GPⅡb-Ⅲa)receptor antagonist(tirofiban).Methods Four 24-month-old Beagle bitches were selected and numbered 1,2,3,and 4 respectively,and their pups were removed by cesarean section in two batches 1-2 days before the expected date of delivery.Bitches 1 and 2 were the first batch.Eighteen newborn pups were removed after cesarean section as the control group.They were divided into three subgroups:1-hour subgroup,4-hour subgroup,and 12-hour subgroup according to postnatal time point,with 6 pups in each subgroup.The newborn pups were injected with normal saline 10 mL/kg via jugular vein immediately after birth.Bitches 3 and 4 were the second batch.Nineteen newborn pups were removed by cesarean section as tirofiban group.They were also divided into three subgroups:1-hour subgroup(n=6),4-hour subgroup(n=6),and 12-hour subgroup(n=7)according to the postnatal time point.The newborn pups were injected with tirofiban hydrochloride injection 10 mL/kg(10 mL injection including 2.5 mg of tirofiban)via jugular vein immediately after birth.The diameter of ductus arteriosus was measured by echocardiography.Ductus arteriosus was removed by surgical dissection and divided into two parts.Western blotting and immunohistochemistry were used to detect the expression of platelet membrane GPⅡb-Ⅲa,respectively.Results In the control group,1 newborn pup died at 0.5 hour after birth in the 1-hour subgroup.The experiment was completed by 19 in the tirofiban group.Ductus arteriosus of all pups were not closed in 1-hour subgroups of the two groups,and there was no significant difference in the diameter of ductus arteriosus between the control group and the tirofiban group(mm:1.72±0.08 vs.1.70±0.11,P>0.05).Ductus arteriosus of 1 newborn pup in 4-hour subgroup of the control group was closed,but the ductus arteriosus of all the newborn pups in 4-hour s

关 键 词：血小板膜糖蛋白类 血小板 动物 新生 动脉导管 

分 类 号：R722.1[医药卫生—儿科]