重组人促红细胞生成素对家兔心肺复苏后心脏功能影响的实验研究
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摘要
目的复苏后心功能障碍是导致复苏后早期高死亡率的主要原因。本实验目的在于探讨重组人促红细胞生成素(recombinant human erythropoietin,rhEPO)对心肺复苏后家兔心脏功能的影响,明确是否减轻复苏后心肌损伤,改善复苏后心功能障碍,并对其其可能机制进行研究。
方法通过体外致颤法建立家兔心脏骤停心肺复苏模型。18只清洁级健康家兔随机分为三组,A组(n=6),手术对照组,只行麻醉、手术、气管插管,但不致颤；B组(n=6)：肾上腺素组,在复苏时使用标准剂量肾上腺素(30μg/kg)；C组(n=6)：肾上腺素+rhEPO组,在B组基础上,ROSC后5min给予rhEPO(5000IU/kg)。动态观察并记录致颤前,ROSC即刻、15min、30min、60min、120min的左室舒张末压(LVEDP)、左室内压上升和下降最大速率(peak±dp/dt)的变化。复苏后120min处死家兔,检测心肌细胞凋亡和胞内凋亡诱导因子(apoptosis inducing factor,AIF)表达,同时计算心肌细胞凋亡指数(apoptosis index,AI)和AIF表达百分比,并行心肌超微结构检查。
结果1)A组家兔在复苏前和复苏后各观察时间点LVEDP和peak±dp/dt的变化差异均无统计学意义(P均>0.05)；2)与A组比较,B、C两组在复苏成功后各观察点LVEDP均增加(P<0.05或P<0.01),复苏后60min,两组LVEDP呈下降趋势,C组虽较B组明显,但直到120min时才有统计学差异(P<0.01)；3)与A组比较,复苏后B、C两组peak±dp/dt均呈下降趋势(P<0.05或P<0.01),但复苏后60min,C组出现回升,且持续到120min时(P<0.01)；4)三组心肌细胞AI分别为(6.4±2.3)%,(31.2±2.4)%和(12.0±3.2)%。B组AI显著大于C组,差异有统计学意义(P<0.01)；5)三组细胞AIF表达百分比分别为(7.4±2.7)%,(30.2±1.8)%和(18.5±4.7)%,B组显著大于C组,差异有统计学意义(P<0.01)；6)电镜观察显示B、C两组均见心肌细胞肿胀,线粒体及肌丝等超微结构损伤,但以C组损伤较轻。
结论在对室颤导致的心脏骤停进行复苏时,rhEPO可以改善复苏后血流动力学指标,并可抑制复苏后心肌细胞AIF表达及凋亡,减轻心肌组织超微结构损伤,改善复苏后心功能障碍。
Objective Postresuscitation myocardial dysfunction contributes to the high fatality rate following successful resuscitation. The purpose of this animal research was to observe the effects of recombinant human erythropoietin (rhEPO) on postresuscitation cardiac function in the rabbits and further explore the underlying mechanism by which rhEPO exert protective effects on ischemia reperfusion injury myocardium.
Methods After setting up rabbit model of cardiopulmonary resuscitation,18 rabbits were randomly divided into three groups,group A:operation-control group, only anesthesia, surgery, endotracheal intubation, but was not induced ventricular fibrillation; group B:epinephrine group, administration of the standard dose of epinephrine (30μg/kg) during CPR; group C:epinephrine and rhEPO group, on the base of group B,administration of rhEPO(5000IU/kg)after restoration of spontaneous circulation (ROSC) 5 minutes. The left ventricular end-diastolic pressure (LVEDP) and peak first derivative of left ventricular pressure (±dp/dt) were observed. The ischemia reperfusion-induce apoptosis and the apoptosis inducing factor(AIF) were detected by TUNEL and the immunochemical method respectively,and compared the apoptosis index(AI) and the percent of the AIF each groups.The ultrastructure of myocytes was observed by electron telescope.
Results①The LVEDP and peak±dp/dt of group A before and after resuscitation in the recovery of all observation time points were no significant differences(P>0.05).②Compared with group A, the LVEDP of the remaining groups gradully increased in ROSC immediately(P<0.05 or P<0.01), but decreased after ROSC60th,whereas the decreasing in group C was lower than in group B at the time of ROSC 120th(P<0.01).③Compared with group A,the peak±dp/dt of the remaining two groups gradully decreased in ROSC immediately(P<0.05 or P<0.01), whereas the peak±dp/dt in group C began to increase in ROSC 60th until ROSC 120th(P<0.01).④The AIs in three groups were (6.4±2.3)%,(31.2±2.4)%and(12.0±3.2)%,respectively.And compared with each of groups were all having the significant differences(P<0.01).⑤The percentages in three groups were (7.4±2.7)%,(30.2±1.8)%and(18.5±4.7)%,respectively.And compared with each of groups were all having the significant differences(P<0.05or P<0.01).⑥The ultrastructure in group B was protected better than those of group C.
Conclusions rhEPO administered after resuscitation improved postresuscita-tion hemodynamics. It inhibited the expressing of apoptosis inducing factor,the cardiocyte apoptosis and alleviated cardiac ultrastructure damage at the early stage of cardiopulmonary resuscitation after CPR from cardiac arrest,and imporoved postresuscitation myocardial dysfunction.
方法通过体外致颤法建立家兔心脏骤停心肺复苏模型。18只清洁级健康家兔随机分为三组,A组(n=6),手术对照组,只行麻醉、手术、气管插管,但不致颤；B组(n=6)：肾上腺素组,在复苏时使用标准剂量肾上腺素(30μg/kg)；C组(n=6)：肾上腺素+rhEPO组,在B组基础上,ROSC后5min给予rhEPO(5000IU/kg)。动态观察并记录致颤前,ROSC即刻、15min、30min、60min、120min的左室舒张末压(LVEDP)、左室内压上升和下降最大速率(peak±dp/dt)的变化。复苏后120min处死家兔,检测心肌细胞凋亡和胞内凋亡诱导因子(apoptosis inducing factor,AIF)表达,同时计算心肌细胞凋亡指数(apoptosis index,AI)和AIF表达百分比,并行心肌超微结构检查。
结果1)A组家兔在复苏前和复苏后各观察时间点LVEDP和peak±dp/dt的变化差异均无统计学意义(P均>0.05)；2)与A组比较,B、C两组在复苏成功后各观察点LVEDP均增加(P<0.05或P<0.01),复苏后60min,两组LVEDP呈下降趋势,C组虽较B组明显,但直到120min时才有统计学差异(P<0.01)；3)与A组比较,复苏后B、C两组peak±dp/dt均呈下降趋势(P<0.05或P<0.01),但复苏后60min,C组出现回升,且持续到120min时(P<0.01)；4)三组心肌细胞AI分别为(6.4±2.3)%,(31.2±2.4)%和(12.0±3.2)%。B组AI显著大于C组,差异有统计学意义(P<0.01)；5)三组细胞AIF表达百分比分别为(7.4±2.7)%,(30.2±1.8)%和(18.5±4.7)%,B组显著大于C组,差异有统计学意义(P<0.01)；6)电镜观察显示B、C两组均见心肌细胞肿胀,线粒体及肌丝等超微结构损伤,但以C组损伤较轻。
结论在对室颤导致的心脏骤停进行复苏时,rhEPO可以改善复苏后血流动力学指标,并可抑制复苏后心肌细胞AIF表达及凋亡,减轻心肌组织超微结构损伤,改善复苏后心功能障碍。
Objective Postresuscitation myocardial dysfunction contributes to the high fatality rate following successful resuscitation. The purpose of this animal research was to observe the effects of recombinant human erythropoietin (rhEPO) on postresuscitation cardiac function in the rabbits and further explore the underlying mechanism by which rhEPO exert protective effects on ischemia reperfusion injury myocardium.
Methods After setting up rabbit model of cardiopulmonary resuscitation,18 rabbits were randomly divided into three groups,group A:operation-control group, only anesthesia, surgery, endotracheal intubation, but was not induced ventricular fibrillation; group B:epinephrine group, administration of the standard dose of epinephrine (30μg/kg) during CPR; group C:epinephrine and rhEPO group, on the base of group B,administration of rhEPO(5000IU/kg)after restoration of spontaneous circulation (ROSC) 5 minutes. The left ventricular end-diastolic pressure (LVEDP) and peak first derivative of left ventricular pressure (±dp/dt) were observed. The ischemia reperfusion-induce apoptosis and the apoptosis inducing factor(AIF) were detected by TUNEL and the immunochemical method respectively,and compared the apoptosis index(AI) and the percent of the AIF each groups.The ultrastructure of myocytes was observed by electron telescope.
Results①The LVEDP and peak±dp/dt of group A before and after resuscitation in the recovery of all observation time points were no significant differences(P>0.05).②Compared with group A, the LVEDP of the remaining groups gradully increased in ROSC immediately(P<0.05 or P<0.01), but decreased after ROSC60th,whereas the decreasing in group C was lower than in group B at the time of ROSC 120th(P<0.01).③Compared with group A,the peak±dp/dt of the remaining two groups gradully decreased in ROSC immediately(P<0.05 or P<0.01), whereas the peak±dp/dt in group C began to increase in ROSC 60th until ROSC 120th(P<0.01).④The AIs in three groups were (6.4±2.3)%,(31.2±2.4)%and(12.0±3.2)%,respectively.And compared with each of groups were all having the significant differences(P<0.01).⑤The percentages in three groups were (7.4±2.7)%,(30.2±1.8)%and(18.5±4.7)%,respectively.And compared with each of groups were all having the significant differences(P<0.05or P<0.01).⑥The ultrastructure in group B was protected better than those of group C.
Conclusions rhEPO administered after resuscitation improved postresuscita-tion hemodynamics. It inhibited the expressing of apoptosis inducing factor,the cardiocyte apoptosis and alleviated cardiac ultrastructure damage at the early stage of cardiopulmonary resuscitation after CPR from cardiac arrest,and imporoved postresuscitation myocardial dysfunction.
引文
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3.叶任高,钟南山.内科学(第七版)[M].北京：人民卫生出版社,2008：229-232.
4.王一镗,沈洪.心肺脑复苏(第二版)[M].上海：上海科学技术出版社,2007：20-21.
5.王其新.现代临床急救进展[M].北京：人民军医出版社,2003：1-20.
6. The American Heart Association.2005 American Heart Association Guidelines for Cardiapulmonary Resuscitation and Emergency Cardiovascular Care [J]. Circulation,2005,112(24Suppl):IV 19-20.
7. Mary F,Vinay M,Robert W,et al.Major changes in the 2005 AHA Guideline for CPR and ECC reaching the tipping for change[J].Circulation,2005,114(4):206-211.
8.胡辉莹,钟世镇.心肺复苏中胸外按压作用及研究进展[J].中国急救医学,2006,26(12)：928-930.
9. Wanchun Tang,方向韶.心肺脑复苏研究的现状和对策[J].中华急诊医学杂志,2007,16(1)：7-9.
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