ABSTRACT
Objective To evaluate the delivery of recombinant adeno-associated virus(rAAV)-mediated CD151 gene in promoting neovascularization and coronary collateralization in swines after myocardial infarction.Method Twenty-six chinese minipigs(clean,provided by breeding pig farm of Huazhong Agricultural University) were randomly divided into four groups:(1)normal control group(n=4):swines without surgery.(2)rAAV-GFP group(n=7):The acute myocardial infarction models were produced in swines by ligating the left anterior descending coronary artery(LAD).The success criteria of the models was that ST-segment elevations in leatds I and aVL maintained for 20 minutes.The rAAV-GFP was injected into the left ventricular anterior wall(divided into 10 points,1×10 11 pfu/point).(3)rAAV-CD151 group(n=7):The operation method was the same as rAAV-GFP group.The rAAV-CD151 was injected into the left ventrieular anterior wall(divided into 10 points).(4) rAAV-antiCD151 group(n=8):The operation method was the same as rAAV-GFP group.The rAAV-antiCD151 was injected into the left ventricular anterior wall(divided into 10 points).Eight weeks after coronary artery liga-tion,the expression of CD151 was measured by western blot.Coronary angiography was done to evaluate collateral circulation of the infarct zone of myocardium.The infurct size was determined by staining with triphenyl-tetrazolium chloride(TTC).Statistical analysis wan carried out by using one-way analysis of variances.Results High level of CD151 protein expression was detected.Coronary angiography showed better collateral circulation in the rAAV-CD151 group.The percentage of infarct size was sinificanly lower in the rAAV-CD151 group (12.82±2.26)% than that in the other two groups,and that was higber in rAAV-anti-CD151 group(32.52±3.47)% than that in the rAAV-GFP group(23.14%±2.83%,both P＜0.05).Conelusions CD151 in vivo gene transferred to swines with acute myocardial infarction promotes neovascularization and thereby improves collateral circulation.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the efficacy of CD151 gene delivery in promoting blood perfusion in swines after myocardial infarction.</p><p><b>METHODS</b>Swines received coronary artery ligation and intramyocardial injection with rAAV-CD151, rAAV-anti-CD151 or rAAV-GFP. Eight weeks after vector injection, Western blot, immunostaining and 13N-labeled NH3 PET were performed to detect gene expression and biological effects of various treatments.</p><p><b>RESULTS</b>High level of CD151 protein expression was detected in the rAAV-CD151 group. The capillary density in the rAAV-CD151 group [(83.8 +/- 6.7) n/mm2] was significantly higher than that in the control group [(33.2 +/- 4.5) n/mm2] and rAAV-GFP group [(41.6 +/- 5.6) n/mm2] (all P<0.05); the arteriole density in the rAAV-CD151 group [(16.4 +/- 2.5) n/mm2] was also higher than that in the control group [(6.6 +/- 2.3) n/mm2] and the rAAV-GFP group [(8.4 +/- 1.6) n/mm2] (all P<0.05). However, the lowest capillary density and arteriole density were evidenced in rAAV-anti-CD151 group. Myocardial blood perfusion was significantly increased in rAAV-CD151 group and significantly reduced in rAAV-anti-CD151 group (all P<0.05 vs. control).</p><p><b>CONCLUSION</b>Intramyocardial injection of rAAV-CD151 could enhance the myocardial express of CD151 protein, increase capillary and arteriole densities and improve blood perfusion in swine with myocardial infarction.</p>
Subject(s)
Animals , Female , Humans , Male , Antigens, CD , Genetics , Coronary Occlusion , Therapeutics , Dependovirus , Genetics , Gene Transfer Techniques , Genetic Therapy , Genetic Vectors , Myocardial Infarction , Therapeutics , Neovascularization, Physiologic , Swine , Swine, Miniature , Tetraspanin 24 , Treatment OutcomeABSTRACT
Objective To investigate the effect of CD151 gene therapy on improving myocardial function in swines with myocardial infarction. Methods CD151, antisense CD151 and green fluorescent protein (GFP) were constructed into the recombinant adeno-associated virus (rAAV). Swines were divided into 4 groups: rAAV-GFP group (6 swines), rAAV-CD151 group (6 swines), rAAV-antiCD151 group (6 swines) and control group (6 swines). The swines were performed with coronary artery ligation and intramuscularly injection with rAAV. Eight weeks after vector administration, western blot was used to detect gene expression of CD151. 13N-labeled NH3 positron emission tomography (PET) was used to evaluate myocardial perfusion. Echocardiography was used to assess myocardial function. Results Compared with the control group and the rAAV-GFP group, the rAAV-CD151 group showed higher CD151 protein expression. Compared with the rAAV-GFP group, the defect size of myocardium was decreased[( 11.3±2.4)% vs. (21.1±2.6)%, t= -5.67,P<0.01] and left ventricular ejection fraction (EF), left ventricular fractional shortening (FS), the ratio of anterior lateral wall thickening (△ALWT) and ratio of interventricular septum thickening (△IVST) were significantly improved in rAAV-CD151 group 8 weeks after vector administration [(65.7±4.6)% vs. (54.7±5.3)%, (36.0±2.9)% vs. (27.6±3.1)%,(55.4± 4.9)% vs. (36.8±7.8)%, (35.2±6.0)% vs. (26.7±4.4)%, t=3.98, 3.35, 3.34, 9.27, all P< 0.05]. The level of diastolic ALWT and diastolic IVST was also increased in rAAV CD151 group compared with rAAV-GFP group ( P<0.05).Compared with rAAV-CD151 group, parameters of myocardial function in rAAV-antisense CD151 group were not improved (P<0.05). Conclusions rAAV-CD151 can effectively transfeet the myocardium, increase the expression ofCD151 protein, promote the blood perfusion of myocardium and improve the ventricular function after myocardial infarction.
ABSTRACT
Objective: The signal transduction pathway of phosphatidylinositol-3-kinase(PI3K) may play important roles in promoting angiogenesis induced by growth factors.The involvement of CD151,a member of transmember-4 superfamily,remains unclear.This study aimed to investigate the effects of the recombinant adeno-associated virus mediated CD151(rAAV-CD151) gene delivery on capillary density in myocardial infarction swine and its mechanisms.Methods: We established the swine model of myocardial infarction by coronary artery ligation,and intramuscularly injected rAAV-CD151,rAAV-GFP and rAAV-antiCD151 into the ischemic myocardium.Eight weeks after coronary artery ligation,we detected the expression of CD151,and measured capillary density by immunostaining for the von Willebrand factor.Results: Compared with the control and the rAAV-GFP groups,the rAAV-CD151 group showed a higher CD151 protein expression and capillary density in ischemic myocardium after myocardial infarction(P