Li Dayuan, Liu Yong, Chen Jiawei, Velchala Neelima, Amani Fariba, Nemarkommula Aravind, Chen Kui, Rayaz Hassan, Zhang Dazhi, Liu Hongmei, Sinha Anjan K, Romeo Francesco, Hermonat Paul L, Mehta Jawahar L
Gene Therapy Program, Division of Cardiovascular Medicine, University of Arkansas for Medical Sciences, VA Medical Center, Little Rock, AR 72205, USA.
Biochem Biophys Res Commun. 2006 Jun 9;344(3):701-7. doi: 10.1016/j.bbrc.2006.04.010. Epub 2006 Apr 19.
TGFbeta(1) deficiency has been attributed to the development of atherosclerosis. There is, however, little direct evidence for this concept. To examine this hypothesis, low-density lipoprotein receptor knockout (LDLR(-/-)) mice were injected via tail vein with recombinant adeno-associated virus type 2 (rAAV) carrying a bioactive TGFbeta(1) mutant (AAV/TGFbeta1ACT, n=10) or granulocyte-macrophage-colony stimulating factor (AAV/GM-CSF, n=10, a negative control) or saline (n=9, control), and then put on a high cholesterol diet. At 18 weeks, blood lipids were found to be similarly elevated in all LDLR(-/-) mice. TGFbeta1ACT and GM-CSF (DNA, mRNA, and protein) were highly expressed in the tissues of mice given TGFbeta1ACT or AAV/GM-CSF, respectively, showing sustained transfection following gene delivery by the systemic route. Saline-treated and AAV/GM-CSF-treated LDLR(-/-) mice showed extensive areas of atherosclerotic lesion formation. There was evidence of intense oxidative stress (nitrotyrosine staining), inflammation (CD68 staining), and expression of adhesion molecules and the ox-LDL receptor LOX-1 (gene array analysis) in the atherosclerotic tissues. Importantly, atherosclerotic lesion formation was markedly inhibited in the LDLR(-/-) mice given AAV/TGFbeta1ACT. Expression of adhesion molecules and LOX-1, oxidative stress, and inflammatory response all were inhibited in the mice given AAV/TGFbeta1ACT (P<0.05 vs. saline-treated or GM-CSF-treated LDLR(-/-) mice). These data for the first time demonstrate that systemic delivery of TGFbeta1ACT gene via AAV can inhibit formation of atherosclerotic lesions, possibly via anti-inflammatory and anti-oxidant mechanisms. These findings suggest a novel view of TGFbeta(1) in atherogenesis and a potential new gene therapy for treatment of atherosclerosis.
转化生长因子β1（TGFbeta(1)）缺乏被认为与动脉粥样硬化的发生有关。然而，这一观点几乎没有直接证据。为了验证这一假说，通过尾静脉给低密度脂蛋白受体基因敲除（LDLR(-/-)）小鼠注射携带生物活性TGFbeta(1)突变体的重组腺相关病毒2型（rAAV）（AAV/TGFbeta1ACT，n = 10）或粒细胞-巨噬细胞集落刺激因子（AAV/GM-CSF，n = 10，阴性对照）或生理盐水（n = 9，对照），然后给予高胆固醇饮食。18周时，发现所有LDLR(-/-)小鼠的血脂均同样升高。TGFbeta1ACT和GM-CSF（DNA、mRNA和蛋白质）分别在给予TGFbeta1ACT或AAV/GM-CSF的小鼠组织中高表达，表明经全身途径进行基因传递后实现了持续转染。生理盐水处理和AAV/GM-CSF处理的LDLR(-/-)小鼠出现了广泛的动脉粥样硬化病变形成区域。在动脉粥样硬化组织中有强烈氧化应激（硝基酪氨酸染色）、炎症（CD68染色）以及黏附分子和氧化型低密度脂蛋白受体LOX-1表达（基因芯片分析）的证据。重要的是，给予AAV/TGFbeta1ACT的LDLR(-/-)小鼠的动脉粥样硬化病变形成受到显著抑制。给予AAV/TGFbeta1ACT的小鼠中黏附分子和LOX-1的表达、氧化应激及炎症反应均受到抑制（与生理盐水处理或GM-CSF处理的LDLR(-/-)小鼠相比，P<0.05）。这些数据首次证明，通过AAV进行TGFbeta1ACT基因的全身传递可抑制动脉粥样硬化病变的形成，可能是通过抗炎和抗氧化机制。这些发现提示了TGFbeta(1)在动脉粥样硬化发生过程中的新观点以及治疗动脉粥样硬化的潜在新基因疗法。
