地氟烷后处理及JAK2/STAT3通路在小鼠脑缺血再灌注损伤中的作用  

Post-conditioning with desflurane and role of JAK2/STAT3 pathway in alleviating cerebral ischemia reperfusion injury in mice

作　　者：赵晓艳[1] 胡春华 梁轩[1] 潘楚雄 王古岩 ZHAO Xiao-yan;HU Chun-hua;LIANG Xuan;PAN Chu-xiong;WANG Gu-yan(Department of Anesthesiology,Beijing Tongren Hospial,Capital Medical University,Beijing 100730,China;Department of Anesthesiology,Beijing Stomalological Hospilal,Capital Medical University,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京同仁医院麻醉科,北京100730 [2]首都医科大学附属北京口腔医院麻醉科,北京100050

出　　处：《中华实用诊断与治疗杂志》2021年第9期896-900,共5页Journal of Chinese Practical Diagnosis and Therapy

基　　金：国家自然科学基金(81671044)。

摘　　要：目的观察地氟烷后处理的脑缺血再灌注(ischemia reperfusion,IR)损伤小鼠神经行为学及脑组织病理变化,探讨JAK2/STAT3通路在其中的作用。方法SPF级雄性BALB/c小鼠50只,随机分为IR组、IR+地氟烷组、IR+JAK2抑制剂组、IR+地氟烷+JAK2抑制剂组和假手术组各10只。IR组采用线栓法阻塞大脑中动脉60 min后恢复灌注,制备IR模型;IR+地氟烷组制备IR模型后吸入体积分数8%地氟烷60 min;IR+JAK2抑制剂组在大脑中动脉阻塞前即刻腹腔注射JAK2抑制剂AG49040 mg/kg,余操作同IR组;IR+地氟烷+JAK2抑制剂组在大脑中动脉阻塞前即刻腹腔注射AG49040 mg/kg,缺血处理60 min后恢复灌注,吸入体积分数8%地氟烷60 min;假手术组只进行手术操作,不阻塞大脑中动脉,不吸入地氟烷,不腹腔注射AG490。再灌注24 h后,比较5组小鼠神经行为学评分;然后处死小鼠,取脑组织缺血半影区,行TUNEL染色并计算凋亡细胞数,采用Western blot法检测脑组织JAK2、p-JAK2、STAT3、p-STAT3、Bcl-2、Bax蛋白相对表达量并计算p-JAK2/JAK2、p-STAT3/STAT3。结果IR组[(6.83±0.74)分]、IR+地氟烷组[(4.21±0.52)分]、IR+JAK2抑制剂组[(7.90±1.03)分]、IR+地氟烷+JAK2抑制剂组[(5.02±0.54)分]神经行为学评分均高于假手术组(0分)(P<0.05);IR+地氟烷组神经行为学评分低于IR组(P<0.05);IR+JAK2抑制剂组神经行为学评分高于IR组(P<0.05);IR+地氟烷+JAK2抑制剂组神经行为学评分低于IR+JAK2抑制剂组(P<0.05)。IR组[(48.52±4.36)个]、IR+地氟烷组[(23.04±3.35)个]、IR+JAK2抑制剂组[(59.00±6.14)个]、IR+地氟烷+JAK2抑制剂组[(35.63±4.28)个]脑组织缺血半影区凋亡细胞数均多于假手术组[(5.00±1.00)个](P<0.05);IR+地氟烷组脑组织缺血半影区凋亡细胞数少于IR组(P<0.05);IR+JAK2抑制剂组脑组织缺血半影区凋亡细胞数多于IR组(P<0.05);IR+地氟烷+JAK2抑制剂组脑组织缺血半影区凋亡细胞数少于IR+JAK2抑制剂组(P<0.05)。IR组、IR+地氟烷组、IR+Objective To observe the changes of neurological behaviors and brain histopathological tissues after post-conditioning with desflurane(Des)in mice with cerebral ischemia reperfusion(IR)injury,and to investigate the roles of JAK2-STAT3 pathway.Methods Fifty male SPF BALB/c mice were randomly divided into IR group,IR+Des group,IR+AG490(a JAK2 selective inhibitor)group,IR+Des+AG490 group and sham-operation group,with 10 mice in each group.IR injury models were prepared by reperfusion 24 hafter 60-min middle cerebral artery occlusion(MCAO)in IR group.In IR+Des group,8%Des was inhaled for 60 min after reperfusion.In IR+AG490 group,40 mg/kg AG490 was intraperitoneally injected just before MCAO,and the other procedures were as same as in IR group.In IR+Des+AG490 group,40 mg/kg AG490 was intraperitoneally injected just before MCAO and then 8%Des was inhaled for 60 min after reperfusion.In sham-operation group,the common carotid artery was only exposed without being occluded nor AG490 and Des treated.After 24 hreperfusion,the neurobehavioral score was compared among five groups.Then the mice were sacrificed and the peri-ischemic region of the brain was obtained.TUNEL method was used to calculate the number of apoptotic cells and Western blot was used to detect the relative expressions of JAK2,p-JAK2,STAT3,p-STAT3,Bcl-2 and Bax protein to calculate p-JAK2/JAK2 and p-STAT3/STAT3.Results The neurobehavioral score was higher in IR group(6.83±0.74),IR+Des group(4.21±0.52),IR+AG490 group(7.90±1.03)and IR+Des+AG490 group(5.02±0.54)than that in sham-operation group(0)(P<0.05),lower in IR+Des group than that in IR group(P<0.05),higher in IR+AG490 group than that in IR group(P<0.05),and lower in IR+Des+AG490 group than that in IR+AG490 group(P<0.05).The number of apoptotic cells was greater in IR group(48.52±4.36),IR+Des group(23.04±3.35),IR+AG490 group(59.00±6.14)and IR+Des+AG490 group(35.63±4.28)than that in sham-operation group(5.00±1.00)(P<0.05),smaller in IR+Des group than that in IR group(P<0.05),greater in IR+AG490

关 键 词：脑缺血再灌注损伤 地氟烷 JAK2/STAT3通路 小鼠 

分 类 号：R743.3[医药卫生—神经病学与精神病学]