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时间:2020-06-03
《白介素33对乳鼠心肌细胞乏氧/复氧损伤时胞内活性氧自由基生成的影响.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、中国心血管杂志2014年4月第19卷第2期ChinJCardiovascMed,April2014,Vo1.19,No.2·139·.基础研究.白介素33对乳鼠心肌细胞乏氧/复氧损伤时胞内活性氧自由基生成的影响李冠臻张新超【摘要】目的探讨白介素33(IL一33)对乳鼠心肌细胞乏氧/复氧损伤的保护作用及胞内活性氧自由基(ROS)生成的影响。方法分离培养sD乳鼠心肌细胞,将原代培养的心肌细胞随机分为单纯对照(C)组、加药对照(rIL一33100ng/ml,C1)组、乏氧/复氧(A/R)组、A/R+rII
2、一33(1、10、100ngJm1)组,乏氧培养3h后,复氧2h。免疫组化鉴定心肌细胞;MTY法检测细胞存活率;ELISA试剂盒检测乳酸脱氢酶(LDH)漏出率;流式细胞仪检测细胞凋亡(AV—PI双标法)、胞内ROS生成量(ROS—DHE荧光探针)。结果与c组相比,A/R组心肌细胞存活率明显下降(P<0.01)。LDH漏出显著增高(P<0.叭),凋亡率及胞内ROS含量显著增加(P<0.01);与A/R组相比,IL一33治疗组剂量依赖性地提高细胞存活率(P<0.叭),降低LDH漏出率、细胞凋亡率(P<0
3、.01)及胞内ROS含量(P:0.03);加药对照组与C组相比.心肌细胞存活率、凋亡率、胞内ROS含量差异无统计学意义。结论IL一33在心肌乏氧/复氧损伤过程中可以剂量依赖性地发挥抗细胞凋亡作用.其保护作用机制可能与增强心肌抗氧化能力,减少胞内ROS生成有关。【关键词】白细胞介素33;活性氧;肌细胞;心脏;乏氧/复氧损伤InfluenceofIL-33onROSgenerationinneonatalratsduringanoxia/reoxygenation—inducedmyocardialin
4、juryLiGuanzhen,ZhangXinchao.DepartmentofEmergency,BeijingHospital,MinistryofHealth,Bering100730,ChinaCorrespondingauthor:ZhangXinchao,Email:xinchaoz@163.com【Abstract】ObjectiveToinvestigatetheprotectiveeffectofinterleukin一33(IL一33)onanoxia/reoxygenation
5、(A/R)一inducedinjuryanditsinfluenceonreactiveoxygenspecies(ROS)generationinneonatalrats.MethodsPrimarycardiomyocyteswereisolatedfromneonatalSprague—DawIeyratsandwererandomlydividedinto6groups:controlgroup,control+rIL一33100ng/mlgroup.A/RgroupandA/R+rIL一3
6、3(1,10,100ng/m1)groups.A/RgroupandA/R+rIL一33aresubjectedto3hoursofanoxia,followedby2hoursreoxygenation.Cardiomyocyteswereidentifiedbyimmunocytochemicalstaining.TheviabilityofthecellswasexaminedbyMTTassay.Theleveloflactatedehydrogenase(LDH)wasdetectedby
7、aLDHenzymelinkedimmunosorbentassay(ELISA)kit.Theapoptosisofthecardiomyocyteswasobservedviafloweytometry(AnnexinVandpropidiomiodidedoublestainingmethod),ROSlevelsweremeasuredbyROSfluorescentprobe—dihydr0ethidium(DHE).ResultsComparedwithcontrolgroup,thea
8、poptosisrateofthecardiomyocytes,levelofLDHandROSwereremarkablyincreased(aⅡP<0.01),theviabilityofcardiomyocyteswasimpairedsignificantlyinA/Rgroup(P<0.01).ComparedwithA/Rgroup,theapoptosisrateofthecardiomyocytes(P<0.O1),levelofLDHandROSpr
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