阻断缺血后处理大鼠心肌线粒体比较蛋白质组学的分析

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1、SSP7803(葡萄糖调节蛋白75，Grp75)，表达下降的3个蛋白分别为SSPl609(--氢硫辛酸脱氢酶，丙酮酸脱氢酶复合体组成部分，DLDH)、SSP2107(rCG44606)、SSP3002(ATP合酶，ATPsynthase)。5．WeStemblot验证DLDH、Grp75在两组中的表达情况，在5-HD+IPO组中DLDH较IPO组下降，Grp75较IPO组上升，这些表达变化与2-DE的结果一致且差异有统计学意义俨<0．05)。结论：1．缺血后处理能明显改善大鼠心脏功能，而5．羟葵酸可部分阻断其保护作用；2．5-HD阻断缺血后处理后：@D

2、LDH表达下调，推测缺血再灌注损伤激活PDK(丙酮酸脱氢酶激酶)促使DLDH磷酸化而下调；@ATP合酶表达下调，其原因可能是膜电位的改变致使ATP合酶亚基的旋转过程受损；⑨G印7S表达上调，推测可能是线粒体Ca2+超载、活性氧的生成增加等因素激活了热休克转录因子，促进Grp75的基因转录和蛋白表达；④IMMT表达上调，可能是由于线粒体内一增加、ATP合成障碍、醛类物质堆积等原因，促使蛋白质发生了代偿性的升高；3．差速离心联合Nycodenz不连续密度梯度离心，可获取较高纯度的心肌线粒体。关键词：心肌保护；5．羟葵酸；缺血后处理；线粒体敏感性钾通道；蛋白

3、质组学；双向电泳6ComparativeproteomicsstudyonmyocardialmitochondrialproteinsinratsfollowingantagonizingischemicpOstconditioningAbstractobjective：Tostudymyocardialmitochondrialproteinsexpressioninratsfollowing5-HDantagonizingischemicpostconditioninginallattempttofindtheprotectiveproteins

4、andmarkerproteinsofischemiareperfusion,providingthebasisforbasicandclinicalstudies．Methods1．Langendorffapparatuswereusedtoestablishthemodelofmyocardialischemiareperfusioninjury．24maleSprague—Dawleyratswererandomlydividedinto4groups(n=6，eachgroup)，i．e．thenormalgroup(Nor)，theI／Rgr

5、oup(I／R)，theischemicpostconditioninggroup(IP0)，5-hydroxydecanoateplusischemicpostconditioninggroup(5HD+IPO)．GroupNorwasperfusedwithKerbs-Henseleit(K-H)solutionfor120minuteswithoutproducingischemia-reperfusioninjury．TheotherthreegroupswereperfusedwimK-Hsolutionabout20minutesforeq

6、uilibration．St．Thomassolutionwasperfusedtostoptheheartbeatingafterequilibration，andthentheheartswereunderwent40minutesglobalischemia．GroupI／RwasperfusedwiththeK-Hsolutionfor60minutes．GroupIPO：after40minutesofglobalischemia,heartswerereperfusedfor10seconds，andarrestedfor10seconds

7、for6cycles，followedbyperfusion嘶mK-Hsolutionfor58minutes．Group5-HD+IPOWasperfusedwith5-HD(100lunol／LofK—Hsolution)for3minutesandthenpostconditioningwasadministeredasthatinGroupIPO，followedbyK-Hsolutionfor5minutes．Thecardiacfunctionindexes(suchasFIR、LVDP、LVDEPandtheMaxdp／dt)ofeach

8、groupattheendpointofequilibrationandreperfusion