缺血前预处理对机体内一氧化氮表达的影响

Effects of ischemic pretreatment on expression of nitric oxide in vivo

目的 观察缺血前预处理对机体内一氧化氮（NO）表达的影响.方法 选择成年健康Wistar沙土鼠50只,建立离体Langendorff灌流模型,按照缺血前处理方法的不同平分为5组,正常对照组选择恒压灌流105 min;缺血再灌注组进行平衡20 min,停灌45 min后再灌注60 min;预处理组在停灌45 min后,再灌注60 min前给予6个循环的30 s缺血（R）/30 s再灌注（Ⅰ）;预处理＋依达拉奉组或丹参组使用含依达拉奉或丹参的灌流液于停灌前灌注10 min,并至再灌注结束.结果 正常对照组、缺血再灌注组、预处理组、预处理＋依达拉奉组与预处理＋丹参组在灌注60 min时冠脉流出液中的乳酸脱氢酶（LDH）活性分别为（352.12±9.63）、（615.36±66.36）、（473.19±50.18）、（412.39±35.48）、（399.54±45.89） mU.与正常对照组比较,缺血再灌注组再灌注60 min时冠脉流出液中LDH活性显著升高（P＜0.05）,而预处理能显著降低冠脉流出液中LDH酶活性（P＜0.05）.正常对照组苏木素-伊红（HE）染色显示心肌细胞有序排列,无炎性细胞浸润;缺血再灌注组心肌细胞排列杂乱,细胞肿胀,有少量炎性细胞浸润;预处理3组的细胞损害程度明显低于缺血再灌注组.与正常对照组比较,再灌注60 min后缺血再灌注组心肌组织内NO含量与总一氧化氮合酶（TNOS）和诱导型一氧化氮合酶（iNOS）活性显著升高（P＜0.05）,而预处理能显著降低心肌组织内NO含量与TNOS和iNOS活性（P＜0.05）;依达拉奉与丹参的处理使预处理对心肌组织内上述指标的降低作用更显著（P＜0.05）.结论 缺血前预处理可减少离体培养心肌组织的LDH释放,从而表现出对心肌细胞的保护作用,该作用可能是通过对NO-iNOS信号通路进行调节而实现的,依达拉奉与丹参的应用能更加有效发挥预处理作用.

Objective To investigate the effects of ischemic pretreatment on the expression of nitric oxide （NO） in vivo.Methods Fifty adult healthy Wistar gerbils were selected,there were established in vitro Langendorff perfusion model,accorded to different levels of ischemic pre-treatment methods were equally divided into 5 groups （the normal control group were selected constant perfusion 105 min; ischemia; And the reperfusion group were balance 20 min,45 min after stopping irrigation irrigation 60 min; The pretreatment group were 45 min after stopping irrigation,60 min before reperfusion given six cycles 30 s R/30 s Ⅰ; The pretreatment ＋ edaravone group and the pretreatment ＋ salvia group were used the Salvia edaravone or salvia perfusion perfusion 10 min before stopping irrigation.Results The coronary effluent lactate dehydrogenase （LDH） activity in the six groups were （352.12 ± 9.63）,（615.36 ± 66.36）,（473.19 ± 50.18）,（412.39 ± 35.48）,（399.54 ± 45.89） mU,Compared with normal control group,the coronary effluent LDH activity in the ischemia-reperfusion group 60 min reperfusion were significantly increased （P 〈 0.05）,whereas the pretreatment can significantly reduce coronary effluent LDH activity （P 〈 0.05）.Hematoxylin and eosin （HE） staining showed that the normal control group were ordered myocardial cells and were no inflammatory cell infiltration; The myocardial ischemia-reperfusion group were showed arranged clutter,cell swelling and were small amount of inflammatory cell infiltration; The pretreatment were showed damage were significantly lower than the ischemic reperfusion group.Compared with normal control group,reperfusion within 60 min after myocardial,the NO content and total nitric oxide synthase （TNOS）,inducible nitric oxide synthase （iNOS） activity of the ischemia-reperfusion group were significantly increased （P 〈0.05）,whereas pretreatment can significantly reduce NO content and TNOS iNOS activity in the myocardial tissue （P 〈 0.05） ; The edaravone and salvia treatment can reduce the NO content and TNOS iNOS activity in the myocardial tissue （P 〈 0.05）.Conclusion Ischemic preconditioning can reduce lactate dehydrogenase release in the myocardial tissue,thus it showed the protective effect on myocardial cells,which it may be through the NO-iNOS signaling pathways regulate,Edaravone and Salvia applications can play more effective role in the pretreatment.