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董素明1，常文凯2，罗俊茜1，薛佳杰1，贾英伟2，梁炳生21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030001Ultrastructuralchangesinaratmodeloflowerlimbischemia/reperfusioninjuryundergoingedaravoneDongSu-ming1,ChangWen-kai2,LuoJun-qian1,XueJia-jie1,JiaYing-wei2,LiangBing-sheng21ShanxiMedicalUniversity,Taiyuan030001,ShanxiProvince,China;2DepartmentofOrthopedics,SecondHospital,ShanxiMedicalUniversity,Taiyuan030001,ShanxiProvince,China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40kPa达到阻断血流，4h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5min由左股静脉注射依达拉奉3mg/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24h，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-2mRNA、baxmRNA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24h后，右胫前肌bcl-2mRNA的相对表达量以及bcl-2mRNA与baxmRNA的比值：模型组显著低于依达拉奉灌注组(P<0.05)，而baxmRNA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-2mRNA、抑制baxmRNA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。中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程全文链接：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；bax；氧自由基；Abstract：BACKGROUND:Theoxygenfreeradicalsandapoptosisplayanimportantroleinlimbischemia/reperfusioninjury,sowecanalleviatelimbischemia/reperfusioninjurybyinhibitingtheproductionofoxygenfreeradicalsandapoptosis.OBJECTIVE:Todiscusstheapplicationandeffectofedaravoneonlimbischemia/reperfusioninjuryinrats.METHODS:Ofthe30femaleSprague-Dawleyrats,20ratswererandomlyselectedtomakemodelsoflimbischemia/reperfusioninjurybyligatingtherootofrightlowerlimbwithaself-madeballooncuffat40kPapressuretoblockbloodflowfor4hoursandreperfusing.Aftersuccessmodelestablishment,theywererandomlyassignedtotwogroups.Intheedaravoneperfusiongroup,edaravone3mg/kgwasinjectedviatheleftfemoralveinat5minutesbeforereperfusion.Inthemodelgroupandnormalgroup(theremaining10rats),anequalvolumeofphysiologicalsalinewasgivenatthesametimepoint.At24hoursafterreperfusion,therightanteriortibialmuscleofeachgroupwasremovedandtheseultrastructuralchangeswereobservedbytransmissionelectronmicroscope.Bcl-2mRNAandBaxmRNAofratanteriortibialmuscleofeachgroupweresemiquantitativelydetectedwiththeRT-PCRandtheratioofbcl-2/baxwascalculated.RESULTSANDCONCLUSION:(1)Electronmicroscoperesults:comparedwiththemodelgroup,themusclefiberswereneater;theMlineandtheNlinewereclearer;theswellingofmitochondriawasalleviated;thenumbersofmitochondriaandmitochondrialcristawerealsoincreasedintheedaravoneperfusiongroup.(2)RT-PCRresults:At24hoursafterreperfusion,therelativeexpressionofbcl-2mRNAandtheratioofbcl-2mRNAtobaxmRNAinrightanteriortibialmusclewer...