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1、促红细胞生成素对缺氧/复氧心肌骨架蛋白作用【摘要】目的从大体和细胞的角度,观察促红细胞生成素(EPO)对缺氧/复氧心肌细胞骨架蛋白:肌动蛋白(a-actininprotein)、微管蛋白(tubulinprotein)和结蛋白(desminprotein)的影响,并初步探讨EPO对缺氧/复氧心肌骨架蛋白作用机制。方法经夹闭气管8min建立窒息性大鼠心脏骤停-心肺复苏动物模型。24只Sprague-Dawley大鼠随机(随机数字法)分为3组:健康对照(N)组、全心缺血/再灌注(I/R)组和EPO组。I/R组于窒息致心搏骤停后8
2、min,予胸外按压和机械通气进行复苏,恢复自主循环(ROSC),监测复苏后心功能状态。EPO组CPR后于ROSC后3min注射EPO5000U/kg)。确定模型成功后观察120min,取心室肌,SABC免疫组化法观察微管蛋白、结蛋白和肌动蛋白变化。分离培养SD大鼠乳鼠心肌细胞,建立缺氧/复氧模型。培养心肌细胞随机分为4组:①正常(C)组;②缺氧/复氧(H/R)组(缺氧10h/复氧4h);③EPO组(缺氧10h/复氧4h后予EPO10U/mL干预),免疫荧光法观察各组细胞微管蛋白、肌动蛋白结构及其荧光强度变化。结果动物实验中,
3、N组、I/R组和EPO组结蛋白、微管蛋白和肌动蛋白的免疫组化平均光密度值(0D)组间比较差异均无统计学意义。细胞研究实验中,H/R组心肌细胞肌动蛋白、微管蛋白荧光染色结构明显破坏,网状结构模糊不清,平均荧光强度值较C组明显减弱,但与EPO组比较差异均无统计学意义。结论缺氧/复氧对心肌损伤作用具有时效性,EPO对缺氧/复氧损伤的心肌骨架蛋白无保护作用。【关键词】促红细胞生成素;缺氧/复氧;骨架蛋白;肌动蛋白;微管蛋白;结蛋白Theeffectoferythropoietinonmyocardialcytoskeletalpro
4、teinsafterhypoxia/reoxygenationinjuryJiangHuilin,WangHuajun,LiBingliu,ChenXiaohui,ZhuYongcheng,LinPeiyi・EmergencyDepartment,TheSecondAffiliatedHospitalofGuangzhouMedicalUniversity,Guangzhou510260,ChinaCorrespondingauthor:ChenXiaohui,Email:cxhgzl68@126.com[Abstract]
5、ObjectiveToobservetheimpactofhypoxia/reoxygenationonmyocardialcytoskeletaiproteins(a-actininprotein,tubulinprotein,desminprotein)andtoinvestigateEPOlesseningthedamageofmyocardialcytoskeletonproteinsinratsprovedbyculturinghypoxia/reoxygenationinjuredmyocardialcellsi
6、npresenceofEPO.MethodsTheratmodelofasphyxia-inducedcardiacarrestwasperformedbyturning-offtheventilatorandclampingtheendotrachealtube・Afterasphyxiafor8minutes,CPRwascarriedout.Atotalof24ratsweredividedintonormalgroup,ischemia/resuscitation(I/R)groupandtheEPOgroup(n二
7、8).Themodelofmyocardialdysfunctionwasdetermined2hoursafterrestorationofspontaneouscirculation(ROSC).TheratsofEPOgroupweregivenEPO5000U/kgafterROSC.Theratheartspecimenswerecollected.Actinin,TubulinandDesminproteinwereobservedbySABCimmunohistochemistry.Theculturedcar
8、diomyocytesweretakenfromneonatairatsandweredividedintothreegroups:thenormalgroup,thehypoxia/reoxygenation(H/R)group(hypoxia10h/reoxygenate4h),the