Plasmid-mediated VEGF gene transfer induces cardiomyogenesis and reduces myocardial infarct size in sheep

We have recently reported that in pigs with chronic myocardial ischemia heart transfection with a plasmid encoding the 165 isoform of human vascular endothelial growth factor (pVEGF165) induces an increase in the mitotic index of adult cardiomyocytes and cardiomyocyte hyperplasia. On these bases we hypothesized that VEGF gene transfer could also modify the evolution of experimental myocardial infarct. In adult sheep pVEGF165 (3.8 mg, n=7) or empty plasmid (n=7) was injected intramyocardially 1 h after coronary artery ligation. After 15 days infarct area was 11.3+/-1.3% of the left ventricle in the VEGF group and 18.2+/-2.1% in the empty plasmid group (P<0.02). The mechanisms involved in infarct size reduction (assessed in additional sheep at 7 and 10 days after infarction) included an increase in early angiogenesis and arteriogenesis, a decrease in peri-infarct fibrosis, a decrease in myofibroblast proliferation, enhanced cardiomyoblast proliferation and mitosis of adult cardiomyocytes with occasional cytokinesis. Resting myocardial perfusion (99mTc-sestamibi SPECT) was higher in VEGF-treated group than in empty plasmid group 15 days after myocardial infarction. We conclude that plasmid-mediated VEGF gene transfer reduces myocardial infarct size by a combination of effects including neovascular proliferation, modification of fibrosis and cardiomyocyte regeneration.     

Reduction of infarct size in the rat heart by LPS preconditioning is associated with expression of angiogenic growth factors and increased capillary density.

Inflammation induces the expression of angiogenic growth factors in tissues, which leads to microvascular growth. Bacterial lipopolysaccharide (LPS) provokes a transient inflammatory response in the heart and induces delayed cardiac resistance to post-ischemic contractile dysfunction. In this study, we examined: 1) the effects of LPS on myocardial expression of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), 2) whether an increase in the density of myocardial microvessels follows the expression of angiogenic growth factors, and 3) the effect of LPS on myocardial resistance to infarction and its relationship with microvascular growth. Rats were treated with LPS (from Salmonella typhimurium, 0.5 mg/kg i.p.). The expression of bFGF and VEGF in the myocardium was examined at 6 and 12 h after LPS treatment by immunofluorescent staining. Myocardial capillary and arteriole densities were determined 3 days after LPS treatment by morphometry, using immunofluorescent staining of von Willebrand factor (a marker protein of endothelial cells) and alpha-smooth muscle actin (a marker protein of smooth muscle cells). To examine cardiac resistance to infarction, hearts were subjected to 40 min of regional ischemia and 2 h of reperfusion by reversible occlusion of left coronary artery at 3 days after LPS treatment. LPS induced cardiac bFGF and VEGF at 6 and 12 h after treatment. The expression of these growth factors was followed by an increase in myocardial capillary density (2032 +/- 78/mm2 vs. 1617 +/- 47/mm2 in saline control, P < 0.05), but not arteriole density, at 3 days. Meanwhile, infarct size was significantly reduced by LPS preconditioning (infarct/left ventricle 12.3 +/- 1.04% vs. 21.7 +/- 1.65% in saline control, 43% reduction, P < 0.05). These results suggest that LPS preconditioning induces cardiac bFGF and VEGF, and an increase in myocardial capillary density. This increased myocardial capillary density is associated with a reduced infarct size after in vivo regional ischemia-reperfusion.     

转染血管内皮细胞生长因子基因骨髓间充质干细胞移植对大鼠心肌梗死区血管新生的影响

背景:骨髓间充质干细胞作为基因载体,有利于外源基因保持良好的遗传稳定性,如能将血管内皮细胞生长因子基因转入间充质干细胞内移植到心肌梗死区,可望获得基因和细胞治疗相互协同促进的作用.目的:观察转染血管内皮细胞生长因子基因的骨髓间充质干细胞移植大鼠心肌梗死区血管新生及血管内皮细胞生长因子基因的体内表达.设计、时间及地点:实验于2004/2007年在湖南省儿童医院儿科研究所,中南大学湘雅二医院中心实验室完成的随机对照实验.材料:实验动物为雄性清洁级Wistar近交系大鼠.以冠脉结扎法制备人鼠心肌梗死模型.方法:以密度梯度离心与贴壁培养法获得大鼠骨髓间充质干细胞.当问充质干细胞生长汇合率为80%～90%时以脂转法转染pcDNA3.1-hVEGF165质粒.心肌梗死模型大鼠随机分4组进行干预:联合组于心肌梗死区移植转染血管内皮细胞生长因子基因的间充质干细胞,细胞组移植间充质干细胞、基因组注射脂质体-pcDNA3.1-VEGF165DNA复合物,对照组注射等容积培养液.主要观察指标:移植4周后,各组大鼠心肌梗死区毛细血管密度,血管内皮细胞生长因子基因表达.结果:火鼠心肌梗死区毛细血管密度以联合组和基因组最高,高于细胞组(P=0.001,0.029),细胞组高于对照组(P=0.028).RT-PCR显示血管内皮细胞生长因子基因体内的表达从高到低依次为联合组、基因组、细胞组和对照组.结论:①作为血管内皮细胞生长因子基因的载体,骨髓间充质干细胞有利于其的稳定表达.②转染血管内皮细胞生长因子基因的骨髓间充质干细胞移植有利于大鼠心肌梗死区的血管新生,其疗效优于单独应用基因或细胞治疗.     

Brain natriuretic peptide limits myocardial infarct size dependent of nitric oxide synthase in rats

BACKGROUND: Brain natriuretic peptide (BNP) has recently been shown to have a cardioprotective effect in animal models of myocardial ischemia-reperfusion (I-R) injury. We hypothesized that exogenous BNP limits myocardial infarction on nitric oxide synthase pathway. METHODS: A rat model of myocardial I-R injury was established by ligating the left descending coronary artery for 30 min and then reperfusing for 2 h. BNP was injected with different dose 5 min after the ligation and lasting for 145 min. The myocardial infarct size and the area at risk of ischemia were measured by staining with triphenyltetrazolium chloride (TTC) and Evans blue dye. To examine the role of nitric oxide synthase (NOS), expression of eNOS in the left ventricle was analyzed by western blotting. Nomega-nitro-L-arginine methyl ester (L-NAME; 30 ug/kg), or S-methylisothiourea (SMT; 3 ug/kg) was administrated before I-R with or without BNP. RESULTS: The control infarct-to-risk ratio was 45.1+/-1.72% (means+/-SE). BNP infused 5 min after ischemia limited infarct size in a dosage-dependent manner, with maximal protection observed at 0.01 ug/(kg min) (infarct-to-risk: 24.7+/-1.69%, P<0.01 vs. control), associated with a 10-fold increase of myocardial endothelial nitric oxide synthase above the control value. Protection afforded by BNP was abolished by L-NAME but not by SMT, suggesting the involvement of putative endothelial but not inducible nitric oxide synthase activation. CONCLUSIONS: We conclude that natriuretic peptide/NOS/NO signaling may constitute an important injury-limiting mechanism in myocardium.     

Effect of Terplex/VEGF-165 gene therapy on left ventricular function and structure following myocardial infarction. VEGF gene therapy for myocardial infarction.

BACKGROUND: We used a novel lipopolymeric gene delivery system, TeplexDNA, to transfect myocardium with plasmid vascular endothelial growth factor-165 (pVEGF) and evaluated the ability of pVEGF to preserve left ventricular function and structure after coronary ligation in a rabbit model. METHODS: New Zealand white rabbits underwent circumflex coronary ligation after direct intramyocardial injection of either Terplex alone or Terplex + 50 microg pVEGF-165. Serial echocardiography and histologic studies were performed (n = 12/group). Mortality did not differ between groups. The data is reported as the mean +/- standard deviation. RESULTS: Over the 21 days following coronary ligation, pVEGF-165-treated animals demonstrated significant improvement in fractional shortening (20-25%, p = 0.02), long axis two-dimensional ejection fraction (42-51%, p=0.02) and short axis m-mode ejection fraction (46-54%, p = 0.02). No significant improvements were noted in the control group. VEGF-treated animals had a 50% increase in peri-infarct vessel density and a trend towards a smaller infarct size (20% vs. 29%, p = 0.10). In animals receiving pVEGF-165, the diastolic ventricular area increased from 1.87 +/- 0.24 cm2 prior to ligation to 2.19 +/- 0.23 cm2 at 21 days following ligation, compared to an increase from 1.84 +/- 0.38 to 2.54 +/- 0.55 cm2 over the same period in control animals (p = 0.03). Similarly, the systolic ventricular area in VEGF-165 animals increased from 1.06 +/- 0.26 cm2 prior to ligation to 1.50 +/- 0.29 cm2 at 21 days following ligation, compared to an increase from 1.16 +/- 0.30 to 1.86 +/- 0.43 cm2 over the same period in the control animals (p = 0.04). CONCLUSION: TerplexDNA mediated delivery of plasmid VEGF administered at the time of coronary occlusion improves left ventricular function and reduces left ventricular dilation following myocardial infarction.     

超声二维斑点追踪技术量化评价陈旧性心肌梗死患者左心室整体心肌梗死大小:与磁共振延迟强化成像对照研究

目的 探讨使用2D-STI技术量化评价陈旧性心肌梗死患者左心室整体心肌梗死大小的可行性及有效性.方法 对43例确诊的连续性陈旧性心肌梗死患者行2D-STI、MR延迟强化成像(DE-MRI).2D-STI左心室整体水平指标包括整体纵向应变(GLS)、旋转及扭转指标,而量化评价左心室整体水平心肌梗死大小指标包括整体心肌梗死容积率及梗死质量.结果 在左心室整体水平,GLS与左心室整体心肌梗死容积率显著相关(r=0.620,P=0.008);GLS和心尖最大旋转率是左心室整体心肌梗死容积率的显著预测因子(P=0.005、0.014),标准回归系数(Beta)分别为0.720和0.592;GLS是左心室整体心肌梗死质量的显著预测因子(P=0.024),Beta为0.545.结论 在左心室整体水平,2D-STI技术的左心室GLS可准确量化评价陈旧性心肌梗死患者整体心肌梗死大小,左心室心尖旋转率也能准确评估整体心肌梗死大小.     

Reducible poly(amido ethylenediamine) for hypoxia-inducible VEGF delivery.

Delivery of the hypoxia-inducible vascular endothelial growth factor (RTP-VEGF) plasmid using a novel reducible disulfide poly(amido ethylenediamine) (SS-PAED) polymer carrier was studied in vitro and in vivo. In vitro transfection of primary rat cardiomyoblasts (H9C2) showed SS-PAED at a weighted ratio of 12:1 (polymer/DNA) mediates 16 fold higher expression of luciferase compared to an optimized bPEI control. FACS analysis revealed up to 57+/-2% GFP positive H9C2s. The efficiency of plasmid delivery to H9C2 using SS-PAED was found to depend upon glutathione (GSH) levels inside the cell. SS-PAED mediated delivery of RTP-VEGF plasmid produced significantly higher levels of VEGF expression (up to 76 fold) under hypoxic conditions compared to normoxic conditions in both H9C2 and rat aortic smooth muscle cells (A7R5). Using SS-PAED, delivery of RTP-VEGF was investigated in a rabbit myocardial infarct model using 100 mug RTP-VEGF. Results showed up to 4 fold increase in VEGF protein expression in the region of the infarct compared to injections of SS-PAED/RTP-Luc. In conclusion, SS-PAED mediated therapeutic delivery improves the efficacy of ischemia-inducible VEGF gene therapy both in vitro and in vivo and therefore, has potential for the promotion of neo-vascular formation and improvement of tissue function in ischemic myocardium.     

Moderate normovolaemic haemodilution increases regional myocardial blood flow but has no effect on experimental infarct size

Regional myocardial blood flow was measured with radioactive microspheres before and 1, 3, 6, and 12 h after occlusion of the left anterior descending coronary artery (LAD) in anaesthetized cats. In one group of cats, 1 h after LAD occlusion, moderate normovolaemic haemodilution was started with a dextran 40 solution, which reduced the haematocrit by 20-25% throughout the rest of the experiment. In the other group, the haematocrit was kept at the original level. Infarct size was measured planimetrically 12 h after the coronary ligation, using the tetrazolium staining technique for disclosing the ischaemic myocardium. Haemodilution increased blood flow in the non-ischaemic myocardium and in the so-called 'border zone' comprising both normal, and ischaemic tissue. However, in the severely ischaemic myocardium dependent on collateral blood flow, where there was no admixture of non-ischaemic myocardium, the blood perfusion was not significantly affected by the haemodilution. After 12 h of ischaemia the infarct size in the control group was fully comparable with that in the haemodiluted group: 28 +/- 3 and 27 +/- 4 (percentage of the left ventricle, mean +/- SEM), respectively.     

Echocardiographic evaluation of ventricular remodeling in a mouse model of myocardial infarction.

Gene-targeting in mice is a powerful tool to define molecular mechanisms of ischemic heart disease that determine infarct size, postinfarct left ventricular (LV) remodeling, and arrhythmogenesis. Coronary ligation in mice is becoming a widely used model of myocardial infarction (MI), but the pathophysiologic consequences of MI in mice and its relevance to human MI have not been fully elucidated. To characterize structural and functional changes during evolving MI, we analyzed 2-dimensional-based reconstruction of the left ventricle by noninvasive echocardiography obtained 1 day and 1 week after surgical ligation of the left anterior descending coronary artery in mice. Sequential 2-dimensional short-axis cineloops of the left ventricle were used to measure LV mass, and LV volumes at end-diastole and end-systole. Echocardiographic infarct size was estimated by measuring the volume of akinetic LV segments. Histologic infarct size was measured by planimetry of 9 transverse sections of each heart. There was close correlation between the 2 methods (31% +/- 20% of LV mass and 34% +/- 17% of LV area, respectively; y =.83x + 7.9, r = 0.96, P <.01). LV volumes at end diastole increased significantly between 1 day and 1 week (51 +/- 17 microL vs 78 +/- 46 microL, respectively, P <.05). The relative change in LV volumes at end diastole varied as a function of infarct size (r = 0.93, P <.01). LV mass and the extent of hypertrophy of noninfarcted segments also varied with infarct size (r = 0.92, P <.01; r = 0.90, P <.01, respectively). Thus, echocardiography is an accurate noninvasive tool for determination of infarct size and quantitative characterization of postinfarct remodeling in the mouse model of MI. Alterations in cardiac structure and function after coronary ligation in mice closely resemble pathophysiologic changes in human ischemic heart disease.     

Combined cord blood stem cells and gene therapy enhances angiogenesis and improves cardiac performance in mouse after acute myocardial infarction

BACKGROUND: Gene and stem cell therapies hold promise for the treatment of ischaemic cardiovascular disease. However, combined stem cell and angiogenic growth factor gene therapy for acute ischaemic myocardium has not been previously reported. This study hypothesized that combined stem cell and gene therapy would not only augment new vessels formation but also improve myocardial function in acute ischaemic myocardium. METHODS: Human angiopoietin-1 (Ang1) cDNA and VEGF(165) cDNA were ligated into AAV vector. The purified CD34(+) cells were obtained from human umbilical cord blood samples. Cord blood CD34(+) cells were transduced with AAV vector encoding either the human Ang1 (AAV-Ang1) or VEGF(165) (AAV-VEGF) cDNA alone, or both (AAV-Ang1 plus VEGF). Immediately after ligation of the left anterior descending coronary artery in male SCID mice, culture-expanded CD34(+) cells transduced with AAV-Ang1, AAV-VEGF or AAV-Ang1 plus VEGF were injected intramyocardially at the left anterior free wall. RESULTS: Western blot showed that Ang1 and VEGF protein expressions were enhanced in the CD34(+)cells transduced with AAV-Ang1 and AAV-VEGF, respectively. Infarct size significantly decreased and capillary density significantly increased after treatment with CD34(+)/AAV-Ang1 plus VEGF when compared with treatment by CD34(+) only. Combined therapy with CD34(+) and AAV-Ang1, CD34(+) and AAV-VEGF, CD34(+) and AAV-Ang1 plus VEGF, all showed significantly higher cardiac performance in echocardiography than the therapy with CD34(+) alone 4 weeks after myocardial infarction. CONCLUSIONS: Combined therapy with human umbilical cord blood CD34(+) cells and both Ang1 and VEGF genes reduced infarct size, attenuated the progression of cardiac dysfunction and increased capillary density in acute myocardial infarction in mice.     

Long-term effects of hepatocyte growth factor gene therapy in rat myocardial infarct model

We investigated the long-term effects of human hepatocyte growth factor (HGF) gene therapy in a rat myocardial infarct model. Treatment adenovirus coexpressing the HGF therapeutic gene and the human sodium/iodide symporter (NIS) reporter gene or control adenovirus expressing the NIS gene alone were injected directly into the infarct border zone immediately after permanent coronary ligation in rats (n=6 each). A uniform disease state was confirmed in the acute phase in terms of impaired left ventricular (LV) function by cine magnetic resonance imaging (MRI), large infarct extent by (99m)Tc-tetrofosmin single-photon emission computed tomography (SPECT) and successful gene transfer and expression by (99m)TcO(4)(-) SPECT. After a 10-week follow-up, repeated cine MRI demonstrated no significant difference in the LV ejection fraction between the time points or groups, but a significantly increased end-diastolic volume from the acute to the chronic phase without a significant difference between the groups. Capillary density was significantly higher in the treatment group, whereas arteriole density remained unchanged. Two-photon excitation fluorescence microscopy revealed extremely thin capillaries (2-5?μm), and their irregular networks increased in the infarct border zone of the treated myocardium. Our results indicated that single HGF gene therapy alone induced an immature and irregular microvasculature.     

Effects of positive end-expiratory pressure in an experimental model of acute myocardial infarct in wistar rats

ABSTRACT: Our purpose in this study was to access the pulmonary effects of mechanical ventilation with positive end-expiratory pressure (PEEP; 10 cmH2O) or without PEEP (zero PEEP-ZEEP) in a rat model of acute myocardial infarction that resulted in hypotension but not in pulmonary congestion. METHODS: Wistar rats were anesthetized (1.5% isoflurane) and myocardial infarct was induced by ligature of the anterior interventricular coronary artery. Rats with myocardial infarct were compared with sham-operated (Sham) and closed thorax groups. RESULTS AND CONCLUSION: There was a significant decrease in MAP in the acute myocardial infarct group (92.5 +/- 4.2 mmHg) when compared with closed chest group (113.0 +/- 4.4 mmHg). There was no significant difference between acute myocardial infarct and Sham groups in PEEP or ZEEP. Mechanical ventilation for 120 min resulted in a significant increase in respiratory system elastance in the groups ventilated with ZEEP (2.59 +/- 0.17 and 2.32 +/- 0.17 cmH2O.mL, Sham and acute myocardial infarct groups, respectively). This effect of mechanical ventilation was not observed in the presence of PEEP in both groups. There was no significant increase in the amount of perivascular pulmonary edema measured in all groups studied. Mean airspace linear intercept and lung tissue distortion index also did not show statistically significant difference between Sham and acute myocardial infarct groups. We conclude that in this experimental model of acute myocardial infarct (12.4 +/- 4.1% area of necrotic tissue and 26.4 +/- 4.0% area of ischemic tissue), there was a protective pulmonary effect of PEEP.     

The treatment of acute myocardial infarct with perlinganit

As many as 43 patients with myocardial infarction were under observation. There were 30 men and 13 women aged 36 to 82 years. Of these, 26 patients had large-focal and 17 transmural myocardial infarction. Perlinganit was injected in the form of 0.1% solution by means of a drug-dosing apparatus (DDA-1). The drug infusion lasted 51.1 +/- 4.8 h on the average, the mean infusion rate was 74.5 +/- 3.4 micrograms/min. After 24 hours it was required that in 32 patients the mean infusion rate be increased 1.5-fold. The hemodynamics was assessed with the aid of tetrapolar chest rheography. Well-defined clinical and hemodynamic effects were revealed. In some patients, the clinical signs of left ventricular insufficiency could be eliminated. Contractile function of the left ventricle returns to normal within the first hours of disease. During perlinganit treatment, the antianginal and antiarrhythmic effects were recorded. The data obtained point to the fact that the use of perlinganit for the treatment of acute myocardial infarction is advisable.     

Global longitudinal strain measured by two-dimensional speckle tracking echocardiography is closely related to myocardial infarct size in chronic ischaemic heart disease

2D-STE (two-dimensional speckle tracking echocardiography) is a novel echocardiographic modality that enables angle-independent assessment of myocardial deformation indices. In the present study, we tested whether peak systolic epsilon(parallel) (longitudinal strain) values measured by 2D-STE could identify areas of MI (myocardial infarction) as determined by CE MRI (contrast-enhanced magnetic resonance imaging). Conventional echocardiographic apical long-axis recordings were performed in 38 patients, 9 months after a first MI. Peak systolic epsilon(parallel) measured by 2D-STE in 16 left ventricle segments was compared with segmental infarct mass and transmurality assessed by CE MRI. Segmental values were averaged to global and territorial values for assessment of global function and myocardial function in the coronary distribution areas. CE MRI identified transmural infarction in 27 patients, and a mean infarct size of 36+/-25 g. Peak systolic epsilon( parallel) correlated with the infarct mass at the global level (r=0.84, P<0.001). A strain value of -15% identified infarction with 83% sensitivity and 93% specificity at the global level and 76% and 95% at the territorial level, and a strain value of -13% identified transmural infarction with 80% sensitivity and 83% specificity at the segmental level. Global infarct mass correlates with the wall motion score index (r=0.70, P<0.001), and left ventricular ejection fraction measured by MRI or echocardiography (r=-0.71 and -0.58, both P<0.001). In chronic infarction, peak systolic epsilon(parallel) measured by 2D-STE correlates with the infarct mass assessed by CE MRI at a global level, and separates infarcted from non-infarcted tissue. Global strain is an excellent predictor of myocardial infarct size in chronic ischaemic heart disease.     

Spontaneous delayed recovery of perfusion after thrombolyzed acute myocardial infarction: is it predictable before discharge?

In patients with thrombolyzed acute myocardial infarction, early assessment of the final infarct size is difficult because spontaneous recovery of perfusion and function of the left ventricle may be delayed. This study was undertaken to evaluate the ability of predischarge low-dose dobutamine echocardiography to predict late spontaneous recovery of perfusion assessed by single-photon emission computed tomography after acute myocardial infarction. We prospectively studied 53 consecutive patients with myocardial infarction treated with thrombolysis. Low-dose dobutamine echocardiography and resting (99m)Tc-sestamibi single-photon emission computed tomography (MIBI SPECT) were performed 4 +/- 2 days after infarction. A follow-up SPECT study was carried out in 45 patients after 6 months. Myocardial recovery was defined as a reduction of SPECT defect size by more than 10% at follow-up compared with the early study. In 25 of the 45 patients, the size of the left ventricular perfusion defect decreased significantly from 42% +/- 16% to 27% +/- 10% (group 1), whereas in the remaining 20 patients it showed no significant change (group 2). Predischarge low-dose dobutamine echocardiography showed a significant improvement in wall motion score index compared with baseline in group 1, from 1.62 +/- 0.28 to 1.41 +/- 0.24, P <.001, whereas in group 2 this index remained without significant change. Predischarge low-dose dobutamine echocardiography is an accurate tool for prediction of late recovery of myocardial perfusion after acute myocardial infarction treated with thrombolysis.     

Basic fibroblast growth factor caused no change in collateral flow or infarct size of acutely-infarcted myocardium in rats

The extent to which local administration of basic fibroblast growth factor (bFGF) increased regional myocardial blood flow (Qm) to acutely-infarcted areas of the heart, thereby mediating myocardial salvage, was examined in this study. Myocardial infarction was induced in two groups of rats by ligation of the left coronary artery. The bFGF group (n = 16) received 100 microg bFGF in physiological saline by intramyocardial injection into the infarcted area, while the control group (n = 7) received only saline. The rats were then maintained for four weeks. Among the controls, Qm decreased in the infarcted areas to 6.5+/-6.7% of that in the noninfarcted areas immediately after coronary ligation, then increased to 11.5+/-8.6% during the four-week maintenance period. In the bFGF group, Qm immediately decreased to 17.5+/-14.7% following ligation and remained stable thereafter (18.3+/-9.1%). There were no significant differences between the bFGF and control groups with respect to Qm, the number of viable myocardial cells or the extent of myocardial fibrosis. In this study we failed to show any significant effect of bFGF on coronary angiogenesis in acutely-ischemic myocardium in rats. Application of bFGF using different dosage, different routes of administration, measuring new capillaries morphologically will be needed to confirm the present negative results.     

Steroid administration after myocardial infarction promotes early infarct expansion. A study in the rat.

Whether steroids lead to thinner scars and larger aneurysms by delaying collagen deposition or worsening infarct expansion before significant collagen deposition begins is unknown. Rats underwent either transmural infarction by left coronary ligation or sham operation. Both infarct and sham rats were randomized to methylprednisolone 50 mg/kg i.p. X 4 or saline treatment within 24 h after operation. Sacrifice occurred before (3 d) or after (7 d) collagen deposition typically begins. Despite similar infarct size, infarct wall thickness was 1.35 +/- 0.08 mm in the saline and 0.99 +/- 0.12 mm in the methylprednisolone group (P less than 0.001) at 3 d. This decrease in wall thickness was explained by a decrease in the number of myocytes across the infarct wall (r = 0.99; P less than 0.001), suggesting that steroids promote myocyte slippage. Furthermore, methylprednisolone caused no further infarct thinning or cavity dilatation beyond 3 d. Thus, high-dose methylprednisolone given within 24 h after transmural infarction worsens infarct expansion before collagen is laid down by promoting the slippage of necrotic myocytes.     

Reduction of infarct size in a rat model of regional myocardial ischemia and reperfusion by the synthetic peptide DAHK

OBJECTIVE: The increased mobilization of iron and copper during ischemia is reported to contribute to postischemic injury. DMI-4983 is a small synthetic peptide, Asp-Ala-His-Lys (DAHK), that mimics the high-affinity copper-binding site of the N-terminus of human albumin. This peptide was reported to inhibit copper-induced formation of reactive oxygen species in vitro, reduce interleukin-8 formation in cultured endothelial cells, and improve left ventricular function after global ischemia and reperfusion in the isolated blood-perfused heart of the rat. The aim of this study was to investigate the effects of DMI-4983 on myocardial infarct size caused by regional ischemia and reperfusion in vivo. DESIGN: Rats were subjected to regional myocardial ischemia (25 mins) followed by reperfusion (2 hrs). Randomized groups received either vehicle or DMI-4983 administered as a 5, 10, or 20 mg/kg intravenous bolus along with a 10 mg/kg/hr continuous infusion starting just before reperfusion and continuing throughout the experiment. Infarct size was determined at the end of the experiment. SETTING: Basic research institute and trauma research laboratory. SUBJECTS: Anesthetized male Wistar rats. MEASUREMENTS AND MAIN RESULTS: The area at risk of infarction and hemodynamic variables were similar in all groups. Rats treated with vehicle resulted in an infarct size of 64% +/- 3% of the area at risk of infarction. Intravenous administration of DMI-4983 reduced infarct size to 52% +/- 3%, 50% +/- 2%, and 45% +/- 3% of the area at risk of infarction for 5, 10, and 20 mg/kg, respectively (p < .05 compared with vehicle for each dosage). CONCLUSIONS: Intravenous DMI-4983 administered before the onset of reperfusion and continuously throughout the reperfusion period caused a significant reduction in tissue necrosis in this in vivo model of regional myocardial ischemia and reperfusion, suggesting that DMI-4983 may represent a novel approach for the treatment of myocardial ischemia and reperfusion injury.     

Assessment of infarct size and myocardial function in mice using transesophageal echocardiography.

BACKGROUND: Because transthoracic echocardiography (TTE) has significant limitations in assessing changes consequent to myocardial infarction (MI) in mice, we studied two novel methods to characterize such infarcts. METHODS: Large MIs were produced by proximal left coronary artery ligation, and small MIs by distal left coronary artery ligation. Serum cardiac troponin I levels were measured 24 hours postoperatively. At 2 weeks, mice underwent transesophageal echocardiography (TEE) and TTE. Infarct sizes were determined histologically. RESULTS: Surviving mice were classified according to infarct size. TEE identified all histologically proven large infarcts, and 4 of 5 small infarcts. TTE identified 4 of 5 large infarcts, but only 1 of 5 small infarcts. TEE-derived fractional area change, but not TTE-estimated left ventricular fractional shortening, was significantly different among large, small, and sham infarcts. Cardiac troponin I showed excellent correlation with infarct size and mortality. CONCLUSIONS: Cardiac troponin I was found to predict infarct size and mortality, whereas TEE proved superior to TTE in determining infarct size and/or myocardial function in a murine MI model. These tools should provide more accurate assessments in preclinical studies of ischemic cardiomyopathy.     

Antagonism of leukotriene B4 receptors does not limit canine myocardial infarct size

Injection of leukotriene (LT)B4 (0.1-3 micrograms/kg i.v.) in normal anesthetized dogs produced dose-related leukopenia that was accompanied by arterial hypotension and tachycardia at higher tested doses. LTD4 (0.1-3 micrograms/kg i.v.), in contrast, increased arterial blood pressure, lowered cardiac rate and produced little change in arterial blood leukocyte count. Continuous infusion of LY255283 [(1-(5-ethyl-2-hydroxy-4-(6-methyl-6-(1H-tetrazol-5-yl)- heptyloxy)phenyl)ethanone] (0.33 mg/kg/min i.v.), a selective LTB4 receptor antagonist, resulted in near complete inhibition of leukopenic, hypotensive and tachycardic responses to LTB4 (3 micrograms/kg i.v.) challenge over a 6-hr test period. Persistent antagonism of canine LTB4 receptors was associated with high circulating levels of LY255283 that were bound extensively to plasma proteins. In subsequent experiments, myocardial infarct size was measured following 1 hr of occlusion of the circumflex coronary artery and 5 hr of reperfusion in control dogs infused with vehicle, and in dogs receiving LY255283 (0.33 mg/kg/min i.v.). Drug and vehicle were infused continuously beginning 15 min before coronary artery occlusion. LY255283 treatment essentially did not alter base-line cardiovascular parameters or myocardial oxygen demand when alterations were compared to time-related changes observed in control dogs. LY255283 infusion also did not alter the degree of myocardial ischemia or the intensity and duration of cardiac arrhythmias associated with coronary artery occlusion and reperfusion. Resultant infarct sizes were 43 +/- 5% of the left ventricle placed at risk in control dogs and 32 +/- 5% in dogs given LY255283; this difference was not statistically significant. The extent of left ventricle placed at risk was similar between groups.(ABSTRACT TRUNCATED AT 250 WORDS)