肢体缺血后处理联合乌司他丁对大鼠脑缺血再灌注脑水肿的缓解作用研究

Study on Remission Effects of Limb Ischemia Postconditioning Combined with Ulinastatin on Brain Water Content Induced by Cerebral Ischemia/Reperfusion Injury in Rats

目的:探讨肢体缺血后处理联合乌司他丁对大鼠脑缺血再灌注脑水肿的缓解作用。方法:选取健康雄性成年SD大鼠300只作为研究对象,随机分为假手术组(Sham)、缺血再灌注组(IR)、乌司他丁组(Ul)、肢体缺血后处理组(Lp)、肢体缺血后处理联合乌司他丁组(Lp Ul)。采用线栓闭塞大脑中动脉法建立大鼠局灶性脑缺血再灌注模型;乌司他丁于脑缺血2 h后再灌注即刻尾静脉注射给药;肢体缺血后处理在脑缺血1.5 h后进行,即结扎5 min,再灌注5 min,重复3次。测定五组大鼠神经功能缺损评分、脑梗死体积分数、脑水肿程度、脑组织水通道蛋白4的表达、脑组织细胞凋亡等指标。结果:IR、Ul、Lp和Lp Ul组大鼠的神经功能缺损评分、脑梗死体积分数均依次减少,且两两比较,差异均有统计学意义(P<0.05)。与Sham组大鼠比较,IR、Ul、Lp和Lp Ul组大鼠缺血侧脑含水量均明显增加,差异均有统计学意义(P<0.05);IR、Ul、Lp和Lp Ul组大鼠组内脑缺血2 h后再灌注6、12、24 h时缺血侧脑含水量的差异均有统计学意义(P<0.05)。与Sham组大鼠比较,IR、Ul、Lp和Lp Ul组大鼠缺血侧脑组织水通道蛋白4的表达均明显增加,差异均有统计学意义(P<0.05)。与IR组大鼠比较,Ul、Lp、Lp Ul组大鼠的凋亡细胞阳性率均明显降低,差异均有统计学意义(P<0.05)。结论:肢体缺血后处理联合乌司他丁治疗,可减轻大鼠脑缺血再灌注脑水肿。

OBJECTIVE： To probe into the remission effects of limb ischemia postconditioning combined with ulinastatin on brain water content induced by cerebral ischemia/reperfusion injury in rats. METHODS： 300 healthy male SD rats were randomly selected to be divided into sham operation group（ sham）,ischemia reperfusion group（ IR）,ulinastatin group（ Ul）,limb ischemia postconditioning group（ Lp） and limb ischemia postconditioning combined with ulinastatin group（ Lp Ul）. The focal cerebral ischemia/reperfusion rat model was set up by using the middle cerebral artery occlusion method. After cerebral ischemia of 2 h,ulinastatin was administrated by instillation in tail vein injection; the ischemic postconditioning was conducted after ischemia of 1. 5 h,namely,ligation of 5 min with instillation of 5 min,for repetition of three times. The neurological deficit score,cerebral infarction volume,brain water content,Aquaporin-4（ AQP4） expression and neuron apoptosis were detected. RESULTS： The neurological deficit score,cerebral infarction volume decreased gradually in IR,Ul,Lp and Lp Ul group,with statistically significant difference between two groups（ P〈0. 05）. Compared with Sham group,the ischemic side of brain water content in IR,Ul,Lp and Lp Ul group increased significantly,with statistically significant difference（ P〈0. 05）. There was statistically significant difference among IR,Ul,Lp and Lp Ul group in ischemic side of brain water content with instillation of 6 h,12 h and 24 h after ischemia of 2 h（ P〈0. 05）. Compared with Sham group,the AQP4 expression in IR,Ul,Lp and Lp Ul group increased significantly,with statistically significant difference（ P〈0. 05）. Compared with IR group,the positive rate of apoptosis cells in IR,Ul,Lp and Lp Ul group decreased significantly,with statistically significant difference（ P〈0. 05）. CONCLUSIONS： Limb ischemia postconditioning combined with ulinastatin can relieve the cerebral injury induced by cerebral ischemia reperfusion.