1-磷酸鞘氨醇受体2在大鼠心脏停搏复苏后脑损伤中的作用

Role of shingosine 1-phosphate receptor 2 in brain injury after cardiac arrest and resuscitation in rats

目的评价1-磷酸鞘氨醇受体2(S1PR2)在大鼠心脏停搏复苏后脑损伤中的作用。方法清洁级健康雄性SD大鼠45只,体重280~320 g,8~10周龄,采用随机数字表法分为3组(n=15):假手术组(S组)、心脏停搏复苏组(CAR组)和S1PR2拮抗剂JTE013组(JTE组)。采用经食道心脏起搏法建立大鼠心脏停搏模型,并进行心肺复苏。复苏即刻,JTE013组腹腔注射10 mg/ml JTE-0131 ml;S组和CAR组腹腔注射等容量DMSO。分别于复苏后24和48 h时采用条带移除实验评价神经功能;于复苏后48 h时处死大鼠,检测脑含水量和血脑屏障通透性,采用Western blot法检测皮层组织S1PR2和occludin的表达水平,明胶酶谱法检测皮层组织基质金属蛋白酶-9(MMP-9)的活性,电镜下观察血脑屏障超微结构。结果与S组比较,CAR组和JTE组复苏后24和48 h时条带移除时间延长,脑含水量和血脑屏障通透性增加,皮层组织S1PR2表达上调,MMP-9活性升高,occludin表达下调(P<0.05);与CAR组比较,JTE组复苏后48 h时条带移除时间缩短,脑含水量和血脑屏障通透性降低,皮层组织S1PR2表达下调,MMP-9活性降低,occludin表达上调(P<0.05)。结论S1PR2参与了大鼠心脏停搏复苏后脑损伤过程。

Objective To evaluate the role of sphingosine 1-phosphate receptor 2(S1PR2)in brain injury after cardiac arrest and resuscitation in rats.Methods Forty-five clean-grade healthy male Sprague-Dawley rats,weighing 280-320 g,aged 8-10 weeks,were divided into 3 groups(n=15 each)using a random number table method:sham operation group(group S),cardiac arrest resuscitation group(group CAR),and S1PR2 antagonist JTE013 group(group JTE).Cardiac arrest was induced by transoesophageal cardiac pacing,and cardiopulmonary resuscitation was performed.Immediately after resuscitation,10 mg/ml JTE0131 ml was intraperitoneally injected in JTE013 group,and the equal volume of DMSO was intraperitoneally injected instead in S group and CAR group.The nerve function was evaluated by tape removal test at 24 and 48 h after resuscitation.Rats were sacrificed at 48 h after resuscitation,the brain water content and blood-brain barrier permeability were measured,the expression of S1PR2 and occludin in the cortical tissue was detected by Western blot,the activity of matrix metalloproteinase-9(MMP-9)was detected by gelatin zymography method,and the ultrastructure of blood-brain barrier was observed with an electron microscope.Results Compared with group S,the time to removal of the tape was significantly prolonged at 24 and 48 h after resuscitation,the brain water content and permeability of blood brain barrier were increased,the expression of S1PR2 in cortical tissues was up-regulated,the activity of MMP-9 was increased,and the expression of occludin was down-regulated in CAR and JTE groups(P<0.05).Compared with group CAR,the time to removal of the tape was significantly shortened at 48 h after resuscitation,the brain water content and permeability of blood brain barrier were decreased,the expression of S1PR2 in cortical tissues was down-regulated,the activity of MMP-9 was decreased,and the expression of occludin was up-regulated in group JTE(P<0.05).Conclusion S1PR2 is involved in the brain injury after cardiac arrest and resuscitation in rats.