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《AMPK信號(hào)通路在七氟醚預(yù)處理增強(qiáng)大鼠心肌細(xì)胞缺氧復(fù)氧時(shí)抗氧化能力中的作用.pdf》由會(huì)員上傳分享,免費(fèi)在線閱讀,更多相關(guān)內(nèi)容在行業(yè)資料-天天文庫(kù)。
1、746中華麻醉學(xué)雜志2014年6月第34卷第6期ChinJAnesthesiol,June2014,Vo1.34,No.6·重癥醫(yī)學(xué)·AMPK信號(hào)通路在七氟醚預(yù)處理增強(qiáng)大鼠心肌細(xì)胞缺氧復(fù)氧時(shí)抗氧化能力中的作用趙建力焦麗媛張彥清王峰李耀李靜靜李博劉保江【摘要】目的探討單磷酸腺苷激活蛋白激酶(AMPK)信號(hào)通路在七氟醚預(yù)處理增強(qiáng)大鼠心肌細(xì)胞缺氧復(fù)氧時(shí)抗氧化能力中的作用。方法采用隨機(jī)數(shù)字表法,將大鼠胚胎心肌細(xì)胞(H9C2細(xì)胞)分為3組(n=12):對(duì)照組(C組)、缺氧復(fù)氧損傷組(H/R組)和七氟醚預(yù)處理(sP組);采用隨機(jī)數(shù)字表法,將AMPK基因敲除(AMPKsiRNA)的H9C2細(xì)胞分為3
2、組(n=12):對(duì)照組(A—C組)、缺氧復(fù)氧損傷組(A—H/R組)和七氟醚預(yù)處理(A.sP組)。H9C2細(xì)胞置于95%N,+5%C02培養(yǎng)箱孵育14h,之后置于95%0:+5%CO培養(yǎng)箱中孵育3h,建立缺氧復(fù)氧模型。sP組H9C2細(xì)胞培養(yǎng)液中加入七氟醚,終濃度為2.0mmol/L,每次孵育15min,共3次,間隔15min進(jìn)行七氟醚預(yù)處理。七氟醚預(yù)處理結(jié)束后15min建立缺氧復(fù)氧模型。復(fù)氧3h時(shí),采用lucigenin增強(qiáng)化學(xué)發(fā)光法和DHE染色測(cè)定超氧陰離子水平,并測(cè)定LDH釋放量和cagpase一3活性。結(jié)果與c組比較,H/R組和sP組H9C2細(xì)胞LDH釋放量增多,caspase一3
3、活性增強(qiáng),超氧陰離子水平升高(P<0.O1);與H/R組比較,sP組H9C2細(xì)胞LDH釋放量減少,easpase.3活性減弱,超氧陰離子水平降低,A.H/R組H9C2細(xì)胞LDH釋放量增多,caspage.3活性增強(qiáng),超氧陰離子水平升高(P<0.01);與sP組比較,A—sP組H9C2細(xì)胞LDH釋放量增多,caspase一3活性增強(qiáng),超氧陰離子水平升高(P<0.01);與A—H/R組比較,A—sP組H9C2細(xì)胞LDH釋放量減少,caspase一3活性減弱,超氧陰離子水平降低(P<0.O1)。結(jié)論AMPK信號(hào)通路參與了七氟醚預(yù)處理增強(qiáng)大鼠心肌細(xì)胞缺氧復(fù)氧時(shí)的抗氧化能力,但并未起主要作用?!?/p>
4、關(guān)鍵詞】蛋白激酶類;麻醉藥,吸入;心肌再灌注損傷;抗氧化劑;缺血預(yù)處理RoleofAMPKsignalingpathwayinsevofluranepreconditioning-inducedenhancementofantioxidantcapacityduringhypoxia-reoxygenationincardiomyocytesofratsZhaoJianli,JiaoLiyuan,ZhangYanqing,Wangng,LiYao,LiJingfing,LiBo,LiuBaojiang.DepartmentofAnesthesiology,F(xiàn)imtHospitalofS
5、hanxiMedicalUniversity,Taiyuan030001,ChinaCorrespondingauthor:LiuBaojiang.Email:baojiangliu@163.com【Abstract】ObjectiveToinvestigatetheroleof5'-AMP.a(chǎn)ctivatedprotein(AMPK)signalingpathwayinsevofluranepreconditioning—inducedenhancementofantioxidantcapacityduringhypoxia—reoxygenation(H/R)inthecardio
6、myocytesoftherats.MethodsTheratembryoniccardiomyocytes(H9C2cells)wererandomlydividedinto3groups(n=12each)usingarandomnumbertable:controlgroup(groupC),H/Rgroup,andsevofluranepreconditioninggroup(groupSP).AMPKgeneknockout(AMPKsiRNA)H9C2cellswererandgmlydividedinto3groups(n:12each)usingarandomnumbe
7、rtable:controlgroup(groupA—C),A—H/Rgroup,andsevofluranepreconditioninggroup(groupA-SP).Thecellswereexposedto95%N2+5%CO2inanincubatorfor14hfollowedbyreoxygenationwith95%O2+5%CO2for3h.InSPgroup,thecellswerepreconditionedwith15