前列腺素E_1/目标温度控制联合干预措施对心肺复苏后大鼠微血栓形成抑制效应的初步探讨

Preliminary Study of Inhibitory Effect of Prostaglandin E1 Combined with Target Temperature Management on the Micro-thrombogenesis of ROSC Rats

目的探讨前列腺素E1(PGE1)和目标温度控制(TTM)联合策略对心跳骤停后成功复苏大鼠的微血栓广泛形成的抑制效应。方法采用经食道交流电致颤方式,建立大鼠心肺复苏模型。设立空白对照组(B组,n=14),另将56只成功复苏的恢复自主循环(ROSC)大鼠随机分为4组:ROSC对照组(R组,n=14)、PGE1干预组(P组,n=14)、TTM干预组(T组,n=14)和PGE1/TTM联合干预组(PT组,n=14)。分别在0.5、4、8h3个时间点,对每组5只大鼠进行血小板计数、凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)、纤维蛋白原(Fib)、血栓调节蛋白(TM)和D-dimer的ELISA检测。每个时间点,每组处死3只大鼠,进行心肌组织苏木精伊红(HE)染色以及血管内皮钙黏蛋白(VE-cadherin)mRNA的PCR检测。实验结束时,对每组处死的大鼠进行心肌组织VE-cadherin/血管内皮生长因子受体(VEGFR)荧光双染。结果 PGE1、TTM及其联合干预措施可以不同程度地减少心肌组织微血栓的形成,缓解微血管内皮细胞VE-cadherin蛋白的破坏,并在复苏后0.5h有效抑制VE-cadherin mRNA表达水平的上升(P<0.05),且以PT组更为明显。3种干预方式都能减缓复苏后血浆TM和D-dimer水平的陡然上升(P<0.05),且联合干预措施较单独干预措施的抑制效应更加突出(P<0.05)。结论PGE1和TTM能够通过不同的抗凝途径和内皮细胞保护效应来改善微血管内血栓的广泛形成,且二者联合治疗可能存在协同作用。

Objective To investigate the inhibitory effect of Prostaglandin E1（PGE1）combined with target temperature management（TTM）on the micro-thrombogenesis of ROSC rat.Methods Ventricular fibrillation（VF）was induced by transoesophageal alternating current was used to establish cardiopulmonary resuscitation（CPR）model in rats.Blank control group was set（group B,n=14）.56 successfully resuscitated rats regained ROSC were divided into 4groups：return of spontaneous circulation（ROSC）group（R group,n=14）,PGE1 intervention group（P group,n=14）,TTM intervention group（T group,n=14）and PGE1/TTM joint intervention group（PT group,n = 14）.Blood platelet count,prothrombin time（PT）,activated partial thromboplastin time（APTT）,fibrinogen（Fib）,thrombomodulin（TM）and D-dimer were evaluated with ELISA using blood samples from 5rats of each group at 0.5,4and 8hours after ROSC.Three rats of every group were sacrificed at each time point to assess hematoxylin and eosin（HE）staining and the vascular endothelial（VE）-cadherin mRNA expression with PCR.In the end,VE-cadherin/ vascular endothelial growth factor receptor（VEGFR）double fluorescent immunohistochemistry staining were also examined.Results PGE1,TTM and PGE1/TTM could significantly diminish the micro-thrombogenesis in the heart tissue,reduce the VE-cadherin protein loss of micro-endothelial cells,and lower the VE-cadherin mRNA expression at 0.5hafter ROSC（P〈0.05）.PT group showed most dramatic effect.All the three interventions brought down the sharp rise of TM and D-dimer levels right after CPR（P〈0.05）,and PGE1/TTM seemed to have better effects on these parameters than the single interventions did（P〈0.05）.Conclusion Both PGE1 and TTM could alleviate the micro-thrombogenesis through different anti-coagulative pathways and the protective effect of the ischemic/reperfusion injury of the endothelium from ROSC rat after CPR separately,while the PGE1/TTM combined intervention might have synergistic better effect than either single one.