国际麻醉学与复苏杂志   2014, Issue (10): 3-3
    
蛋白酶抑制剂对大鼠呼吸心跳骤停后早期复苏脑内能量代谢的初步研究
王晓燕, 李新宇1()
1.成都医学院第一附属医院
Effect of protease inhibitor on brain energy metabolism in rats at theearly stage of cardiopulmonary resuscitation
 全文:
摘要：

目的　探讨蛋白酶抑制剂乌司他丁（ulinastain， UTI）对大鼠呼吸心跳骤停后早期心肺复苏（cardiopulmonary resuscitation， CPR）脑内能量代谢的影响和脑保护作用。　方法　成年雄性SD大鼠30只，按照随机数字表法分为对照组（C组）、生理盐水复苏组（NS组）和UTI复苏组（UTI组），每组10只。各组均行麻醉，气管插管及颈动脉插管。C组不做模型，UTI组及NS组采用窒息法制作大鼠心跳骤停（cardiac arrest， CA）CPR模型。UTI组于自主循环恢复（restoration of spontaneous circulation， ROSC）后2 min内经颈动脉推注注射用UTI（105 U/kg），NS组推注等量的生理盐水。记录各组窒息至CA时间和复苏开始至ROSC时间，监测各组ROSC后1 h内的平均动脉压（mean arterial pressure， MAP）。C组仅行气管插管及颈动脉插管，不进行窒息CA及CPR操作。C组于气管切开置管、颈动脉置管后1 h，NS组和UTI组于ROSC后1 h，经颈动脉取血和快速断头取脑组织。脑组织进行脑组织三磷酸腺苷（adenosine triphosphate， ATP）和乳酸（lactate， LA）含量测定，光镜和电镜下观察海马组织的病理改变。　结果　① 组间MAP基线值差异无统计学意义（P>0.05）。NS组和UTI组ROSC后2、5、30 min MAP差异无统计学意义（P>0.05）。NS组和UTI组MAP在ROSC后2 min较基线值都有显著升高［NS组，（166±11） mmHg vs（143±12） mmHg，P<0.01；UTI组，（170±11） mmHg vs （141±16） mmHg；1 mmHg=0.133 kPa；P<0.01］；ROSC后5 min MAP［NS组，（91±9） mmHg；UTI组，（96±10） mmHg］呈下降趋势，低于基线值（P<0.01）；ROSC后30 min MAP［NS组，（125±12） mmHg；UTI组，（122±13） mmHg］可恢复并趋于稳定，但仍低于基线值（P<0.05）。② 与C组比较，NS组和UTI组大鼠脑组织LA值在ROSC后1 h均明显升高［C组∶NS组∶UTI组，（0.29±0.13） mmol/g prot vs （0.69±0.14） mmol/g prot vs （0.53±0.08） mmol/g prot，P<0.01）］；与NS组比较，UTI组大鼠脑组织LA含量降低（P<0.05）。与C组比较，NS组和UTI组大鼠脑组织ATP含量在ROSC后1 h均明显下降［C组∶NS组，（26±6） μmol/g prot vs （12±5） μmol/g prot，P<0.01；C组∶UTI组，（26±6） μmol/g prot vs （19±8） μmol/g prot，P<0.05）］；UTI组大鼠脑组织ATP较NS组高（P<0.05）。③ NS组脑组织苏木精-伊红（hematoxylin and eosin， HE）染色和尼氏染色及电镜下观察病理学改变较UTI组和C组严重，UTI组病理学改变接近C组。　结论　UTI在一定程度上可能可以改善CPR后1 h脑组织能量代谢，减少乳酸堆积，减轻脑组织病理学损伤。
关键词： 蛋白酶抑制剂；　乌司他丁；　心肺复苏；　脑；　能量代谢
Abstract:

Ｏｂｊｅｃｔｉｖｅ　To explore the effect of protease inhibitor ulinastatin（UTI） on brain energy metabolism and cerebral protection after rats' respiratory and cardiac arrest with early cardiopulmonary resuscitation.　Methods　Thirty adult male SD rats were divided into three groups accroding to the table of random numbers（n=10）: control group（group C）， normal saline cardiopulmonary resuscitation group（group NS） and ulinastatin cardiopulmonary resuscitation group（group UTI）. Anaesthesia， tracheotomy and carotid artery cannulation were performed in all groups. Asphyxial cardiac arrest and resuscitation model performed in group NS and group UTI expect group C. 105 U/kg ulinastatin and the same amount of saline were injected via arteria carotis within 2 min after restoration of spontaneous circulation（ROSC） in group UTI and group NS. Time periods from asphyxiation to cardiac arrest（CA） and from cardiopulmonary resuscitation（CPR） performing to ROSC were recorded， the mean arterial pressure（MAP） was monitored within 1 h after ROSC. One hour after tracheostomy and vascular centesis in group C and 1 h after ROSC in group NS and group UTI， rats were sacrificed to get blood samples from carotid arteries and decapitated to get brain tissue samples. Adenosine triphosphate（ATP） content and lactic acid（LA） content of brain tissue were tested. Pathological changes of hippocampus were observed with light microscope and transmission electron microscope（TEM）.　Results　① The baseline of MAP were no statistical significance among three groups（P>0.05）. There were no statistical significance of MAP between group NS and group UTI when 2， 5 min and 30 min after ROSC（P>0.05）. MAP of 2 min after ROSC in group NS and group UTI were higher than those of baseline ［group NS： （166±11） mmHg vs （143±12） mmHg， P<0.01， group UTI： （170±11） mmHg vs （141±16） mmHg， 1 mmHg=0.133 kPa， P<0.01］， however， MAP of 5 min after ROSC in group NS（91±9） mmHg and group UTI（96±10） mmHg showed downtrend， which were lower than those ofbaseline（P<0.01）. Thirty minutes after ROSC, MAP in group NS（125±12） mmHg and group UTI（122±13） mmHg recovered but still lower than those of the baseline（P<0.05）. ② Comparing to group C， the contents of LA in brain samples of the other two groups were increased significantly at 1 h after ROSC［group C ∶ group NS ∶ group UTI， （0.29±0.13） mmol/g prot vs （0.69±0.14） mmol/g prot vs （0.53±0.08） mmol/g prot， P<0.01］， however， the content of LA was lower in group UTI than that in group NS（P<0.05）. Comparing to group C， the contents of ATP in brain samples of the other two groups were decreased significantly at 1 h after ROSC［group C ∶ group NS， （26±6） μmol/g prot vs （12±5） μmol/g prot， P<0.01， group C ∶ group UTI， （26±6） μmol/g prot vs （19±8） μmol/g prot， P<0.05］， the content of ATP were higher in group UTI than that in group NS（P<0.05）. ③ Observations by light microscope with hematoxylin-eosin（HE） staining and nissl staining and by transmission electron microscope revealed that the pathological changes in group NS were more serious than that of group UTI， and the pathological changes in group UTI were nearly to those of group UTI.　Conclusions　To some extent, the improving of brain energy metabolism, the decrease of lactic acidosis and amelioration of the brain pathological changes one hour after CPR may contribute to the effects of UTI.
Key words: Protease inhibitor；　Ulinastatin；　Cardiopulmonary resuscitation；　Cerebral；　Energy metabolism