Department of Physiology and Pharmacology, The University of Toledo, Toledo, OH 43614, USA.
Hum Gene Ther. 2011 Mar;22(3):271-82. doi: 10.1089/hum.2010.059. Epub 2011 Jan 27.
We examined the hypothesis that vascular and renal dysfunction caused by angiotensin II (Ang II) through increased levels of blood pressure, inflammatory cytokines, and oxidative stress in Sprague-Dawley rats can be prevented by lentiviral-mediated delivery of endothelial heme oxygenase (HO)-1. We targeted the vascular endothelium using a lentiviral construct expressing human HO-1 under the control of the endothelium-specific promoter VE-cadherin (VECAD-HO-1) and examined the effect of long-term human HO-1 expression on blood pressure in Ang II-mediated increases in blood pressure and oxidant stress. A bolus injection of VECAD-HO-1 into the renal artery resulted in expression of human HO-1 for up to 6-9 weeks. Sprague-Dawley rats were implanted with Ang II minipumps and treated with lentivirus carrying either the HO-1 or green fluorescent protein. Renal tissue from VECAD-HO-1-transduced rats expresses human HO-1 mRNA and proteins without an effect on endogenous HO-1. Infusion of Ang II increased blood pressure (p < 0.001) but decreased vascular relaxation in response to acetylcholine, endothelial nitric oxide synthase (eNOS) and phosphorylated eNOS (peNOS) levels, and renal and plasma levels of adiponectin (p < 0.05); in contrast, plasma tumor necrosis factor-α and monocyte chemoattractant protein-1 levels increased. Ang II-treated animals had higher levels of superoxide anion and inducible nitric oxide synthase and increased urinary protein and plasma creatinine levels. Lentiviral transduction with the VECAD-HO-1 construct attenuated the increase in blood pressure (p < 0.05), improved vascular relaxation, increased plasma adiponectin, and prevented the elevation in urinary protein and plasma creatinine in Ang II-treated rats. Endothelial-specific expression of HO-1 also reduced oxidative stress and levels of inflammatory cytokines resulting in increased expression of the anti-apoptotic proteins phosphorylated AKT, phosphorylated AMP-activated protein kinase, peNOS, and eNOS. Collectively, these findings demonstrate that endothelial-specific increases in HO-1 expression attenuate Ang II hypertension and the associated vascular dysfunction that is associated with increases in adiponectin and peNOS and reductions in oxidative stress and levels of inflammatory cytokines.
我们研究了这样一个假设，即血管和肾脏功能障碍是由血管紧张素 II（Ang II）引起的，其机制可能是通过升高血压、炎症细胞因子和氧化应激水平。而 Sprague-Dawley 大鼠的这些功能障碍可以通过慢病毒介导的内皮血红素加氧酶（HO-1）传递来预防。我们使用受血管内皮细胞特异性启动子 VE-cadherin（VECAD-HO-1）调控的慢病毒构建体靶向血管内皮细胞，并研究了长期表达人 HO-1 对 Ang II 介导的血压升高和氧化应激的影响。将 VECAD-HO-1 弹丸式注射到肾动脉中，可使人类 HO-1 的表达持续长达 6-9 周。Sprague-Dawley 大鼠被植入 Ang II 微泵并接受携带 HO-1 或绿色荧光蛋白的慢病毒治疗。转导 VECAD-HO-1 的大鼠的肾组织表达人 HO-1 mRNA 和蛋白，但对内源性 HO-1 没有影响。Ang II 的输注增加了血压（p<0.001），但降低了乙酰胆碱、内皮型一氧化氮合酶（eNOS）和磷酸化 eNOS（peNOS）的血管舒张反应以及肾和血浆中的脂联素水平（p<0.05）；相反，血浆肿瘤坏死因子-α和单核细胞趋化蛋白-1 水平增加。Ang II 处理的动物有更高水平的超氧阴离子和诱导型一氧化氮合酶，并增加了尿蛋白和血浆肌酐水平。用 VECAD-HO-1 构建体的慢病毒转导减轻了血压升高（p<0.05），改善了血管舒张，增加了血浆脂联素，并防止了 Ang II 处理的大鼠中尿蛋白和血浆肌酐的升高。内皮特异性 HO-1 表达的增加还降低了氧化应激和炎症细胞因子水平，导致抗凋亡蛋白磷酸化 AKT、磷酸化 AMP 激活的蛋白激酶、peNOS 和 eNOS 的表达增加。总之，这些发现表明，内皮特异性 HO-1 表达的增加可减轻 Ang II 高血压和相关的血管功能障碍，这与脂联素和 peNOS 的增加以及氧化应激和炎症细胞因子水平的降低有关。
