体外循环血浆对体外培养人脐静脉血管内皮细胞钙池操纵钙通道电.doc

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1、体外循环血浆对体外培养人脐静脉血管内皮细胞钙池操纵钙通道电作者：汪进益，范慧敏，张旭敏，卢蓉，李健，刘中民【摘要】目的探讨经体外循环(cardiopulmonarybypass,CPB)后人体炎性血浆对体外培养的人脐静脉血管内皮细胞(HumanUmbilicalVeinEndothelialCells,HUVECs)的钙池操纵的钙通道电流(store-operatedCa2+currents,Isoc)的影响。方法CPB后血浆采自我科心血管外科手术患者，患者均知情同意。将已培养好的HUVECs按所加入的血浆不同分为4组

2、，每组6例：即组I(加入CPB刚开始Omin时血浆的对照组)、组II(加入CPB后25min的血浆)、组111(加入CPB后50min的血浆)及组IV(加入CPB后50min含左旋精氨酸的血浆)。应用全细胞膜片钳记录技术测定并观察各组HUVECs的Isoc变化。结果在HUVECs上可记录到Isoc；当钳制电位为-100mV时，组I〜组IV中HUVECs的Isoc分别为(-302.38±35.13)pA、(-388.59土58.66)pA、(-577・04±93.69)pA及(-365.42±34.79)pA,各组比较差

3、异有统计学意义(P<；O.05或P<；O.01),其中组IV明显低于组III(P<；0.01)o结论CPB后人体炎性血浆可增强血管内皮细胞的钙通道电流，左旋精氨酸可减轻此作用，这对于进一步深入研究CPB围术期血管内皮细胞钙超载的发生机制和防治具有重要意义。【关键词】体外循环；血管内皮细胞；钙池操纵的钙通道电流；膜片钳技术Abstract：OBJECTIVEToobservetheeffectofinflaminatoryplasmaonstore-operatedCa2+currents(Isoc)ofhu

4、manumbilicalveinendothelialcells(HUVECs)culturedinvitroaftercardiopulmonarybypass(CPB)incardiovascularsurgery.METHODSTheplasmawascollectedfrompatientsofDepartmentofCardiacSurgery,ShanghaiEastHospitalAffiliatedTongjiUniversity・Thepatientsallknewtheprogramandagree

5、dwith・TherandomlyselectedHUVECsweredividedintofourgroupswithsameculturecondition：GroupI(controlgroupaddingtheplasmaafter0minCPB),GroupII(addingtheplasmaafter25minCPB),GroupIII(addingtheplasmaafter50minCPB)andGroupIV(addingtheplasmawithL~arginineafter50minCPB)・Th

6、eIsocofHUVECsineachgroupwasdeterminedbywholecellpatch-clamptechniques・RESULTSThedynamicchangesoftheIsocofthesingleHUVECswasdeterminedbytight-sealwhole-cellpatch-clamptechniques・AtthepotentialofTOOmV,thepeakamplitudeofIsocofHUVECswere(-302.38±35.13)pA,(-388.59±58

7、.66)pA,-(577.04±93.69)pAand(-365.42土34.79)pA,respectivelyinthesegroups・TheIsocwassignificantlylowerinthecontrolgroupthaninothergroups(P<；0.05orP<；0.01).ItwasdistinctlylowerintheGroupIIIthanintheGroupIV(P<；0・01).CONCLUSIONTheinjuryandactivationofvascularend

8、othelialcellscanbewidelyinducedbyCPBwhichcouldmarkedlypotentiatethepeakamplitudeofIsocinadose-dependentmanner,butL-argininecanattenuatethisprocess・Keywords:Cardiopu1m