右美托咪定通过抑制内质网过度应激减轻肺缺血-再灌注小鼠脑损伤  

Dexmedetomidine reduce brain injury induced by lung ischemia/reperfusion in mices through inhibiting excessive endoplasmic reticulum stress response

作　　者：项冰倩 颜王鑫 高慧[1] 楼国强[1] 郝卯林[1] 戴雍月[1] 王万铁[1] Xiang Bingqian Yan Wangxin Gao Hui Lou Guoqiang Hao Maolin Dai Yongyue Wang Wantie(Institute of Ischemia/Reperfusion Injury Research, Wenzhou Medical University, Wenzhou 325035, Chin Department of Anorectal Surgery, Wenzhou People＇ s Hospital, Wenzhou 325000, China)

机构地区：[1]温州医科大学缺血-再灌注损伤研究所,浙江省温州325035 [2]温州市人民医院肛肠外科,浙江省温州325000

出　　处：《中华急诊医学杂志》2017年第11期1260-1267,共8页Chinese Journal of Emergency Medicine

基　　金：浙江省公益技术应用研究项目（2013C33168）;浙江省新苗人才计划项目（2014R413043）;温州市公益性科技计划项目（Y20140652）

摘　　要：目的评价右美托咪定（DEX）通过抑制过度内质网应激反应（ERS）减轻小鼠肺缺血-再灌注（I/R）诱发脑损伤的影响。 方法实验于温州医科大学缺血-再灌注损伤研究所进行。雄性健康SPF级C57BL/6J小鼠50只，体质量20~24 g，8~10周龄，按随机数字表法分为5组（n=10）：假手术组（Sham组）、肺缺血-再灌注损伤组（I/R组）、阿替美唑组（atipamezole,Atip组）、右美托咪定（dexmedetomidine,DEX组）、右美托咪定＋阿替美唑（DA组）。采用小鼠在体左侧肺门夹闭30 min再灌注180 min方法制备肺缺血-再灌注损伤（I/R）模型。Atip组、DEX组、DA组分别在肺门阻断前30 min腹腔注射阿替美唑（250 μg/kg）、右美托咪定（20 μg/kg）、右美托咪定20 μg/kg＋阿替美唑250 μg/kg，其余处理同I/R组。再灌注结束后取脑组织，光镜下观察脑细胞的形态学改变，检测脑组织Caspase3酶活性，TUNEL法检测脑细胞凋亡指数，Western blot、RT-PCR检测脑组织p-JNK、Caspase12、CHOP、GRP78蛋白及mRNA水平。采用SPSS 19.0软件行计量资料分析，多组间比较采用单因素方差分析，以P＜0.05为差异有统计学意义。 结果与Sham比较，其余组光镜下脑组织有明显损伤，Caspase3酶活性、脑细胞凋亡指数、p-JNK、Caspase12、CHOP、GRP78蛋白及mRNA水平均明显升高（P＜0.01）；与I/R、Atip组和DA组比较，DEX组光镜下脑细胞损伤减轻，Caspase3酶活性、脑细胞凋亡指数、p-JNK、Caspase12、CHOP表达均有明显下降，GRP78表达明显升高，差异有统计学意义（P＜0.01）;I/R、Atip组和DA组两两相互比较，光镜下脑组织损伤程度相似，Caspase3酶活性、脑细胞凋亡指数、p-JNK、Caspase12、CHOP蛋白及mRNA水平差异无统计学意义（P＞0.05），GRP78水平DA组明显升高，差异有统计学意义（P＜0.01）。结论右美托咪定预先给药可减轻小鼠肺缺血-再灌注诱发的脑损伤，其机制可能�Objective To investigate the effect of dexmedetomidine （DEX） on the reduction of brain injury induced by lung ischemia/reperfusion （I/R） in mices through inhibiting excessive endoplasmic reticulum stress response （ERS）. Methods Fifty healthy SPF male C57BL/6J mices, weighing 20 -24 g, aged 8 - 10 weeks, were divided into 5 groups （n = 10 each） using a random number table： sham operation group （ sham group）, lung ischemia/reperfusion group （ I/R group）, atipamezole goup （ Atip group）, dexmedetomidine group （DEX group）, dexmedetomidine plus atipamezole group （DA group）. The model of lung I/R injury was established by clamping the left hilum of lung for 30 min followed by reperfusion for 180 min. In Atip, DEX and DA groups, atipamezole 250 μg/kg, dexmedetomidine 20 μg/kg and dexmedetomidine 20μg/kg plus atipamezole 250μg/kg were injected intraperitoneally, respectively, at 30 min before modeling, other procedures were as the same as the I/R group. At 180 min of reperfusion, the animals were sacrificed and the brain tissues were harvested for the observation of morphological changes. The Caspase-3 activity and the apoptosis index of the brain cells were also determined. The levels of protein and mRNA expression of p-JNK, Caspase-12, CHOP and GRP78 in brain tissues were detected by Western blot and RT-PCR. The datas were analyzed using SPSS 19. 0 software and multiple-group comparisons were performed using one-way ANOVA, and P 〈 0. 05 for the difference was statistically significant. Results Compared with the sham group, the Caspase3 activity and brain cell apoptosis index, the protein levels and mRNA expressions of p-JNK, Caspase12, CHOP, GRP78 were significantly increased （P 〈 0. 01 ）, brain tissues had obvious damage in I/R, Atip, DEX and DA groups; compared with I/R, Atip and DA group, brain tissues damage was obvious reduced in DEX group, and the Caspase3 activity, brain cell apoptosis index, the protein levels and mRNA expression of p-JNK, Caspase12, CHOP in DEX

关 键 词：右美托咪定 缺血-再灌注损伤 小鼠 脑 内质网应激 细胞凋亡 CASPASE3 

分 类 号：R614[医药卫生—麻醉学]