Muinck Ebo D De, Nagy Norbert, Tirziu Daniela, Murakami Masahiro, Gurusamy Narasimman, Goswami Shyamal K, Ghatpande Satish, Engelman Richard M, Simons Michael, Das Dipak K
Angiogenesis Research Center, Dartmouth Medical School, Hanover, New Hampshire, USA.
Antioxid Redox Signal. 2007 Apr;9(4):437-45. doi: 10.1089/ars.2006.1501.
PR-39, a proline-arginine-rich angiogenic response peptide, has been implicated in myocardial ischemic reperfusion injury. The present study examined the cardioprotective abilities of PR39 gene therapy. Male C57Bl/J6 mice were randomized to intramyocardial injecton of 10(9) p.f.u. adenovirus encoding PR39 (PR39), FGFR1 dominant negative signaling construct (FGFR1-dn), empty vector (EV), or PR39 adenovirus plus 4 microg of plasmid endcoding a HIF1alpha dominant negative construct (PR39 + HIF1alpha-dn). Seven days later, hearts were subjected to 20 min of ischemia (I) and 2 h. reperfusion (R) ex vivo and aortic and coronary flow, left ventricular developed pressure (LVDP), and LVdp/dt were measured. Myocardial infarct (MI) size and cardiomyocyte apoptosis were measured by TTC staining and TUNEL, respectively. PR39 expression was robust up to 14 days after gene transfer and was absent after EV and FGFR1-dn. Hemodynamics showed no differences at baseline, and heart rate remained unchanged in all groups throughout the experiment. After I-R, hemodynamics remained unchanged in PR39 hearts, but deteriorated significantly in the other groups, except for aortic flow, which remained significantly higher in FGFR1-dn than in EV and PR39 + HIF1alpha-dn (p < 0.05), although it was lower than in PR39 (p < 0.05). MI was 8.7 +/- 0.9 % in PR39, 23.8 +/- 1.1% in FGFR1-dn, 29.9 +/- 2.2% in EV, and 30.8 +/- 2.7 % in PR39 + HIF1alpha-dn (PR39 vs. other groups: p < 0.05; FGFR1-dn vs. EV and PR39 + HIF1alpha-dn: p < 0.05). In PR39, HIF-1alpha protein was higher than in FGFR1-dn and EV. Importantly, cotransfection of HIF1alpha-dn with PR39 completely abolished cardioprotection by PR39. Cardioprotection by PR39 is likely conveyed by protective metabolic and survival responses through HIF1-alpha stabilization and not by angiogenesis, because baseline coronary flow was the same in all groups. Abrogation of FGFR1 signaling conveyed an intermediate degree of cardioprotection.
富含脯氨酸 - 精氨酸的血管生成反应肽PR - 39与心肌缺血再灌注损伤有关。本研究检测了PR39基因治疗的心脏保护能力。将雄性C57Bl/J6小鼠随机分为心肌内注射10⁹ 个感染性颗粒的编码PR39的腺病毒（PR39组）、FGFR1显性负性信号构建体（FGFR1 - dn组）、空载体（EV组）或PR39腺病毒加4微克编码HIF1α显性负性构建体的质粒（PR39 + HIF1α - dn组）。7天后，将心脏离体进行20分钟缺血（I）和2小时再灌注（R），并测量主动脉和冠状动脉血流、左心室舒张末压（LVDP）以及LVdp/dt。分别通过TTC染色和TUNEL检测心肌梗死（MI）面积和心肌细胞凋亡。基因转移后14天内PR39表达强劲，而EV组和FGFR1 - dn组则无表达。血流动力学在基线时无差异，且在整个实验过程中所有组的心率均保持不变。缺血 - 再灌注后，PR39组的血流动力学保持不变，但其他组显著恶化，除了主动脉血流，FGFR1 - dn组的主动脉血流仍显著高于EV组和PR39 + HIF1α - dn组（p < 0.05），尽管低于PR39组（p < 0.05）。PR39组的MI为8.7±0.9%，FGFR1 - dn组为23.8±1.1%，EV组为29.9±2.2%，PR39 + HIF1α - dn组为30.8±2.7%（PR39组与其他组比较：p < 0.05；FGFR1 - dn组与EV组和PR39 + HIF1α - dn组比较：p < 0.05）。在PR39组中，HIF - 1α蛋白高于FGFR1 - dn组和EV组。重要的是，HIF1α - dn与PR39共转染完全消除了PR39的心脏保护作用。PR39的心脏保护作用可能是通过HIF1 - α稳定介导的保护性代谢和存活反应实现的，而非通过血管生成，因为所有组的基线冠状动脉血流相同。FGFR1信号的阻断介导了中等程度的心脏保护作用。
