下肢缺血再灌注损伤模型大鼠接受依达拉奉干预后的超微结构变化  

Ultrastructural changes in a rat model of lower limb ischemia/reperfusion injury undergoing edaravone

作　　者：董素明 常文凯[2] 罗俊茜 薛佳杰 贾英伟[2] 梁炳生[2] 

机构地区：[1]山西医科大学,山西省太原市030001 [2]山西医科大学第二医院骨科,山西省太原市030001

出　　处：《中国组织工程研究》2014年第18期2867-2871,共5页Chinese Journal of Tissue Engineering Research

基　　金：山西省社会发展科技攻关计划项目(20120313021-6);课题名称:新型手术方式及麻醉技术在临床中的应用-依达拉奉灌注+术中临时供血治疗四肢超时限血管损伤的研究~~

摘　　要：背景:肢体缺血再灌注损伤中,氧自由基及细胞凋亡发挥重要的作用,通过抑制氧自由基生成及细胞凋亡,可减轻肢体缺血再灌注损伤。目的:验证依达拉奉在大鼠肢体缺血再灌注损伤方面的应用及疗效。方法:30只雌性SD大鼠,随机取20只,分别用自制气囊袖带环扎大鼠右后肢根部加压至40 kPa达到阻断血流,4 h后松解形成再灌注,制作肢体缺血再灌注损伤模型。造模成功后随机分为2组,依达拉奉灌注组于再灌注前5 min由左股静脉注射依达拉奉3 mg/kg,模型组及剩余10只(正常组)于相同时间点给予等量的生理盐水。于再灌注24 h,取每组大鼠右胫前肌,用透射电镜观察其超微结构变化,并采用RT-PCR对每组大鼠胫前肌中bcl-2 mRNA、bax mRNA进行半定量检测并计算bcl-2/bax比值。结果与结论:①电镜结果显示:依达拉奉灌注组与模型组相比肌纤维较整齐,M线、Z线较清晰,线粒体肿胀减轻、其数量及嵴稍增多。②RT-PCR结果显示,缺血再灌注24 h后,右胫前肌bcl-2 mRNA的相对表达量以及bcl-2 mRNA与bax mRNA的比值:模型组显著低于依达拉奉灌注组(P<0.05),而bax mRNA的相对表达量模型组高于依达拉奉灌注组,且两组均高于正常组(P<0.05)。结果提示自由基清除剂依达拉奉可能通过改善线粒体等超微结构及促进bcl-2 mRNA、抑制bax mRNA的表达来减轻肢体缺血再灌注损伤,这可为肢体缺血再灌注损伤的治疗提供新的选择。BACKGROUND：The oxygen free radicals and apoptosis play an important role in limb ischemia/reperfusion injury, so we can al eviate limb ischemia/reperfusion injury by inhibiting the production of oxygen free radicals and apoptosis. 〈br〉 OBJECTIVE：To discuss the application and effect of edaravone on limb ischemia/reperfusion injury in rats. 〈br〉 METHODS：Of the 30 female Sprague-Dawley rats, 20 rats were randomly selected to make models of limb ischemia/reperfusion injury by ligating the root of right lower limb with a self-made bal oon cuff at 40 kPa pressure to block blood flow for 4 hours and reperfusing. After success model establishment, they were randomly assigned to two groups. In the edaravone perfusion group, edaravone 3 mg/kg was injected via the left femoral vein at 5 minutes before reperfusion. In the model group and normal group （the remaining 10 rats）, an equal volume of physiological saline was given at the same time point. At 24 hours after reperfusion, the right anterior tibial muscle of each group was removed and these ultrastructural changes were observed by transmission electron microscope. Bcl-2 mRNA and Bax mRNA of rat anterior tibial muscle of each group were semiquantitatively＆amp;nbsp;detected with the RT-PCR and the ratio of bcl-2/bax was calculated. 〈br〉 RESULTS AND CONCLUSION：（1）Electron microscope results：compared with the model group, the muscle fibers were neater;the M line and the N line were clearer;the swel ing of mitochondria was al eviated;the numbers of mitochondria and mitochondrial crista were also increased in the edaravone perfusion group. （2）RT-PCR results：At 24 hours after reperfusion, the relative expression of bcl-2 mRNA and the ratio of bcl-2 mRNA to bax mRNA in right anterior tibial muscle were lower in the model group compared with the edaravone perfusion group （P〈0.05）. However, relative expression of bax mRNA was greater in the model group than that in the edaravone perfusion group, which were both higher than the normal g

关 键 词：实验动物 组织构建 依达拉奉 肢体 缺血再灌注损伤 细胞凋亡 bcl-2 氧自由基 

分 类 号：R318[医药卫生—生物医学工程]