PAMAM-D纳米载体介导TFinAODN防治心肌缺血再灌注脂质过氧化损伤的实验研究

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 将100只雄性Lewis大鼠随机等分为5组.除假手术组和缺血再灌注组大鼠静脉分别注入0.9%氯化钠溶液和PAMAM-D外,其余3组分别注入与PAMAM-D相藕联的相应寡核苷酸.分别于缺血90 min和再灌注结束后取5组大鼠的血液检测肌钙蛋白T(TnT)、乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化酶(GSH-PX)的含量.将梗死区及边缘区心肌组织剪下,检测TF基因的转录;另分别检测TF的促凝活性(TF:C)和采用检测TF的抗原含量(TF:Ag).结果 假手术组和其余4组大鼠血液中的TnT、LDH、MDA、SOD和GSH-PX的含量比较,差异有统计学意义(P<0.05或<0.01);而AS/TF防治组与其他3个缺血再灌注组比较,差异有统计学意义(P<0.05或<0.01).假手术组梗死区及边缘区心肌组织中TF基因的转录和表达均弱于其他4组(P<0.01),而AS/TF防治组则较其他3个缺血再灌注组明显减弱(P<0.01).结论 PAMAM-D纳米载体介导的TF AODN通过拮抗组织因子,抑制心肌缺血再灌注过程中的脂质过氧化损伤,对心肌缺血再灌注损伤有明显的保护作用.%Objective To investigate the possible mechanism of polyamidoamine dendrimers (PAMAM-D) mediated antisense oligodeoxynucleotide against tissue factor (TF) in improving myocardial ischemia-reperfusion injury in rats. Methods The 100 male Lewis rats were randomly divided into 5 groups.Except for the rats in sham operation group and ischemia-reperfusion group who were administrated with natural solution and PAMAM-D respectively, the rats in the other three groups received corresponding oligodeoxynucleotide coupled with PAMAM-D. The blood samples were collected at the 90th min of ischemia and the end of reperfusion respectively, and the concentrations of troponin T (TnT), lactate dehydrogenase ( LDH ), malondialdehyde (MDA),superoxide dismutase(SOD) and glutathione peroxidase (GSH-PX) were detected. Myocardial tissues in the infarct and ischemic zone were collected for TF procoagulant activity (TF:C) determination and TF antigen (TF:Ag). Results The concentrations of TnT, LDH, MDA, SOD and GSH-PX were significantly different as compared with those in sham operation group ( P＜0.05 or＜0.01 ) ,however, which in AS/TF intervention group were significantly different as compared with those in the other 3 ischemia -reperfusion injure groups( P＜0.05 or ＜0.01 ). The transcription and expression of TF gene in myocardial tissue of the infarct and ischemic zone were significantly increased in ischemia-reperfusion injure groups ,as compared with those in sham operation group( P＜0.05 or ＜0.01 ), however,which were obviously improved in AS/TF intervention group,as compared with those in the other 3 ischemia-reperfusion injury groups ( P＜ 0.01 ). Conclusion The PAMAM-D mediated TF AODN can relieve the peroxidative injury caused by myocardial ischemia-reperfusion, which has obvious protective effect on myocardial ischemia-reperfusion injury by inhibiting the transcription and expression of TF.