聚酰胺—胺型树枝状高聚物纳米载体介导的组织因子反义寡脱氧核苷酸防治心肌缺血再灌注损伤机制的初步研究  

Preliminary research on the mechanism of polyamidoamine dendrimers nanometer vector mediated antisense oligodeoxynucleotide against tissue factor improving myocardial ischemia-reperfusion injury

作　　者：李春海[1] 桑翠玲[1] 宋树英[1] 周万先 尹俊[3] 李文慧[1] 

机构地区：[1]河北北方学院附属第二医院心内科,河北张家口075100 [2]河北省张家口市第一医院老干部科,075100 [3]汕头大学医学院第二附属医院血液科,515041

出　　处：《疑难病杂志》2011年第7期517-521,共5页Chinese Journal of Difficult and Complicated Cases

基　　金：河北省张家口市科学技术与发展指导性计划资助项目(No.1021108D)

摘　　要：目的研究聚酰胺—胺型树枝状高聚物(PAMAM-D)纳米载体介导组织因子(TF)反义寡脱氧核苷酸防治大鼠心肌缺血再灌注损伤的分子机制。方法分别设计合成针对大鼠TF的反义寡脱氧核苷酸(AS/TF)、正义寡脱氧核苷酸(S/TF)和错配寡脱氧核苷酸(Sc/TF),分别耦联于PAMAM-D纳米载体,构建ODN-PAMAM-D的聚合物。将100只雄性Lewis大鼠随机等分为假手术组、缺血再灌注组、AS/TF防治组、S/TF对照组和Sc/TF对照组。除假手术组和缺血再灌注组大鼠静脉分别注入生理盐水和PAMAM-D外,其余3组大鼠分别注入与PAMAM-D相耦联的相应寡核苷酸。各组大鼠于注射后10h开胸暴露心脏,除假手术组以外的其他4组大鼠于冠状动脉左前降支中1/2处结扎造成心肌缺血。心肌缺血90min后,解除结扎,再灌注3h。假手术组的大鼠于冠状动脉左前降支中1/2处只穿线,不结扎,持续4h 30min。分别于缺血90min和再灌注结束后取各组大鼠的血液检测肌钙蛋白T(TnT)和乳酸脱氢酶(LDH)的含量。并于实验结束后取梗死区及边缘区心肌组织检测TF基因的转录、TF的促凝活性(TF:C)和TF的抗原含量(TF:Ag)以及活性氧簇(ROS)、蛋白酶激活受体-1(PAR-1)和p38有丝分裂原激活蛋白激酶(p38MAPK)的表达。结果心肌缺血90min及再灌注3h后,缺血再灌注组大鼠血液中的TnT和LDH含量均显著高于假手术组,而AS/TF防治组均明显低于其他3个缺血再灌注组(P<0.05);梗死区及边缘区心肌组织中TF基因的转录和表达均强于假手术组,AS/TF防治组则较其他3个缺血再灌注组明显减弱(P<0.01)。流式细胞学检测显示,心肌缺血再灌注3h后,各缺血再灌注组大鼠心肌组织中ROS、PAR-1和p38 MAPK的表达均显著增多,AS/TF防治组则明显低于缺血再灌注组、S/TF对照组和Sc/TF对照组(P<0.05)。结论 TF不仅介导了凝血过程,而且通过激活PAR-1和p38 MAPK诱发了炎性反应,从而加重了心肌组织损伤。TF是心肌�Objective To investigate the possible mechanism of polyamidoamine dendrimers（PAMAM-D） mediated antisense oligodeoxynucleotide against tissue factor（TF） improving myocardial ischemia reperfusion injury.Methods Antisense oligodeoxynucleotide（AS/TF）,sense oligodeoxynucleotide（S/TF） and scrambled oligodeoxynucleotide against TF （Sc/TF） were designed and synthesized respectively,and then were coupled with PAMAM-D nanometer vector to establish ODN PAMAM D polymers.100 male Lewis rats were randomly divided into sham operation group,ischemia reperfusion group, AS/TF intervention group,S/TF control group and Sc/TF control group.Besides rats in sham operation group and ischemia reperfusion group were administrated with natural solution and PAMAM-D respectively,rats in other three groups received corresponding oligodeoxynucleotide coupled with PAMAM-D.10 hours after injection,the chest of each rat was opened with the heart exposed except from those in sham operation group,rats in other 4 groups underwent myocardial ischemia reperfusion by ligaturing left anterior descending coronary artery for 90 min and removing the ligation for 3 hours.In sham operation group, only a thread was put under the left anterior descending coronary artery of each rat,and no ligation was performed and lasted for 4 hours and 30 min.Blood sample was collected after 90 min of ischemia and the end of reperfusion respectively for the concentrations of troponin T（TnT） and lactate dehydrogenase（LDH） determination.At the end of reperfusion,myocardial tissue in the infarct and ischemic area were collected for TF gene transcription detection,TF procoagulant activity（TF：C） determination and TF antigen（TF：Ag） assay.In addition,reactive oxygen species（ROS）,protease activated receptor-1（PAR- 1） and p38 mitogen activated protein kinase（p38 MAPK） were also determined.Results After 90 min of ischemia and the end of reperfusion,concentrations of TnT and LDH increased obviously,while those in AS/TF intervention gr

关 键 词：组织因子 反义寡脱氧核苷酸 心肌 缺血再灌注 活性氧簇 蛋白酶激活受体-1 p38有丝分裂原激活蛋白激酶 大鼠 

分 类 号：R541[医药卫生—心血管疾病]