吸入高浓度氢气对大鼠脑缺血-再灌注损伤的影响  

Effect of inhaling high concentration hydrogen on cerebral ischemia-reperfusion injury in rats

作　　者：万强 李庭庭 肖媛 张昕 孙丽英 张玉荣 李文志[1] 杨万超[1] Wan Qiang;Li Tingting;Xiao Yuan;Zhang Xin;Sun Liying;Zhang Yurong;Li Wenzhi;Yang Wanchao(Department of Anesthesiology,the Second Affiliated Hospital of Harbin Medical University,Harbin 150086,China)

机构地区：[1]哈尔滨医科大学附属第二医院麻醉科,150086

出　　处：《中国脑血管病杂志》2021年第5期303-310,共8页Chinese Journal of Cerebrovascular Diseases

摘　　要：目的研究吸入高浓度氢气对大鼠脑缺血-再灌注损伤的影响及其机制。方法纳入雄性健康无特定病原体(SPF)级Sprague Dawley(SD)大鼠72只,采用随机数字表法进行完全随机化分组,将实验动物随机分为3组,分别为假手术组(S组)、脑缺血-再灌注组(M组)和氢气治疗组(H组),每组24只。S组结扎并离断左侧颈外动脉,M组与H组建立大鼠大脑中动脉栓塞(MCAO)模型,S组、M组大鼠于再灌注前1 h、再灌注后20 h和44 h 3个时间点吸入45 min空气(21%O_(2)-78%N_(2)),H组于再灌注前1 h、再灌注后20 h和44 h 3个时间点吸入45 min高浓度氢气(42%H_(2)-21%O_(2)-37%N_(2))。于再灌注后48 h断头处死大鼠,观察大鼠脑梗死面积及脑组织水肿情况,采用Western blot法检测细胞焦亡相关蛋白含半胱氨酸的天冬氨酸蛋白水解酶1(Caspase-1)及凋亡相关斑点样蛋白(ASC)含量,采用酶联免疫吸附测定(ELISA)方法检测白细胞介素1β(IL-1β)和IL-18水平,并记录大鼠缺血-再灌注后1、3、5、7 d时神经功能评分[改良神经功能缺损评分(mNSS)、Garcia评分]、体质量变化情况以及生存状况。结果缺血-再灌注后48 h,3组大鼠脑梗死面积及脑含水量比较,差异均有统计学意义(F值分别为289.70、95.90,均P<0.01);与M组比较,H组脑梗死面积及脑含水量均明显减少[分别为(30.5±3.0)%比(38.0±1.9)%,(79.1±0.2)%比(83.3±0.6)%;均P<0.05]。3组大鼠缺血-再灌注后48 h脑组织Caspase-1、ASC表达水平均差异有统计学意义(F值分别为107.10、100.70,均P<0.01)。与S组比较,M组脑组织Caspase-1、ASC表达水平均显著增加(均P<0.05);与M组比较,H组脑组织Caspase-1、ASC水平均明显降低(均P<0.05)。3组大鼠缺血-再灌注后48 h后血清中IL-1β和IL-18表达水平差异均有统计学意义(F值分别为65.58、59.36,均P<0.01)。与S组比较,M组IL-1β和IL-18水平均显著上升[分别为(74.2±1.6)ng/L比(58.3±0.6)ng/L,(176.7±11.3)ng/L比(102.9±4.5)ng/L;均P<0.05];与Objective To study the effect of inhaling high concentration hydrogen on cerebral ischemia-reperfusion injury in rats and the mechanism.Methods Seventy-two male Sprague Dawley(SD)rats,using the random number table for complete randomization grouping,are randomly divided into 3 groups:sham operation group(group S),cerebral ischemia-reperfusion group(group M)and hydrogen treatment group(group H),with 24 rats in each group.The left external carotid artery was ligated and disconnected in group S.Middle cerebral artery occlusion(MCAO)models of rats were established in group M and H.Rats in group S and M inhaled air(21%O_(2)-78%N_(2))for 45 minutes at 3 time points 1 hour before reperfusion,20 h and 44 h after reperfusion.In group H,rats inhaled high concentration H_(2)(42%H_(2)-21%O_(2)-37%N_(2))for 45 min at 1 hour before reperfusion,20 h and 44 h after reperfusion.Rats were sacrificed 48 h after reperfusion.The changes of cerebral infarction area and brain edema were observed.Western blot was used to detect the changes of pyroptosis-related protein apoptosis-associated speck-like protein containing a CARD(ASC)and cysteinyl aspartate specific proteinase-1(Caspase-1),and enzyme linked immunosorbent assay(ELISA)was used to detect the levels of interleukin-1β(IL-1β)and IL-18.Neurological function scores(modified neurological severity scores[mNSS],Garcia score),changes in body weight,and survival status of the rats at 1,3,5 and 7 days after ischemia-reperfusion were recorded.Results Forty-eight hours after ischemia-reperfusion,the three groups had significant differences in cerebral infarction area and brain water content(F values were 289.70 and 95.90,both P<0.01);compared with group M,the cerebral infarction area and brain water content in group H were significantly reduced([30.5±3.0]%vs.[38.0±1.9]%,[79.1±0.2]%vs.[83.3±0.6]%,respectively,both P<0.05).The expression levels of Caspase-1 and ASC in the brain tissue of the three groups of rats at 48 h after ischemia-reperfusion were significantly different(F values w

关 键 词：氢气 脑缺血-再灌注 细胞焦亡 炎性因子 

分 类 号：R743.3[医药卫生—神经病学与精神病学]