亚低温对于心肺复苏后大鼠海马神经细胞ROS产生量和Caspase-3 mRNA表达的影响

Influence of hypothermia on the reactive oxygen species and expression of Caspase-3 mRNA of hippocampal neurons on the rats after cardiopulmonary resuscitation

目的观察亚低温(Hypothermia,HT)对心肺复苏(Cardiopulmonary resuscitation,CPR)后大鼠海马神经细胞活性氧(Reactive oxygen species,ROS)产生量和半胱氨酸天冬氨酸蛋白酶3(Caspase-3)mRNA表达的影响。方法 37只健康成年雄性SD大鼠随机分为3组:空白对照组、常温CPR组、亚低温CPR组。CPR组再分2个亚组:即自主循环恢复(Return of Spontaneous Circulation,ROSC)后12h、24h组,除空白对照组为5只大鼠外,余各亚组大鼠均为8只。亚低温CPR组在ROSC后立即行亚低温干预。到达各观察时相点时立即取材,应用流式法检测大鼠海马单细胞悬液中活性氧水平,逆转录-聚合酶链反应(RT-PCR)技术测定海马神经细胞Caspase-3 mRNA的表达水平。结果亚低温CPR组ROSC后12h、24h的ROS产生量和Caspase-3 mRNA表达与相同时相点常温CPR组比较均显著降低(P<0.05)。结论 HT能减少CPR后大鼠神经细胞ROS的产生,并能通过抑制Caspase-3 mRNA表达而使神经细胞凋亡减少。

Objective To observe the influence of hypothermia （HT） on the reactive oxygen species （ROS） and expres- sion of Caspase-3 mRNA of hippocampal neurons on the rats after cardiopulmonary resuscitation（CPR）. Methods 37 healthy adult male SD rats, being divided to 3 groups： blank control group, normothermia CPR group, hypothermia CPR group. Each CPR group then were divided into 2 subgroups,which were 12h and 24h subgroup after restoration of spontaneous circulation （ROSC） ,there were 8 rats in every subgroups except for 5 rats of blank control group. Hypothermia CPR group accepted hy- pothermia intervene immediately after ROSC. On the time of observation, flow cytometry was used to determine the level of ROS of monoplast suspension of hippocampus,reverse transcription-polymerase chain reaction （RT-PCR） was used to determine the level of expression of Caspase-3 mRNA. Results The level of ROS and expression of Caspase-3 mRNA of hypothermia CPR groups were obviously lower than the normothermia CPR group （P〈0.05）. Conclusion HT can reduce the level of ROS of neurons on rats after CPR, and decrease the neuronal apoptosis via the reduction of expression of Caspase-3 mRNA.