Adeno-associated virus serotype 9-mediated overexpression of extracellular superoxide dismutase improves recovery from surgical hind-limb ischemia in BALB/c mice.
Adeno-associated virus serotype 9-mediated overexpression of extracellular superoxide dismutase improves recovery from surgical hind-limb ischemia in BALB/c mice.
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DOI：
10.1016/j.jvs.2011.03.278
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发表时间：
2011-09
影响因子：
4.3
通讯作者：
Annex, Brian H.
中科院分区：
文献类型：
--
作者：
Saqib, Amina;Prasad, Konkal-Matt R.;Katwal, Arabindra B.;Sanders, John M.;Lye, R. John;French, Brent A.;Annex, Brian H.
新生血管是一种生理性修复过程，部分依赖于一氧化氮。细胞外超氧化物歧化酶（EcSOD）是超氧化物的主要清除剂，因此是一氧化氮生物利用度的重要调节剂，从而防止血管功能障碍。我们假设骨骼肌中EcSOD的过度表达将改善后肢缺血的恢复。将从巨细胞病毒（CMV）启动子表达EcSOD或荧光素酶（对照）的腺相关病毒（AAV）载体交叉包装到AAV 9衣壳中，并IM注射到12周龄小鼠的后肢肌肉中（1×1011个病毒基因组（vg）/肢）。然后在IM注射后诱导缺血。在注射后第0、7和14天通过激光多普勒连续测量肢体灌注，并且值表示为相对于非缺血肢体的比率。Western blotting检测EcSOD表达。毛细血管密度通过血小板内皮细胞粘附分子（PECAM）的免疫组织化学染色来记录。通过末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）试验评估细胞凋亡，并每天目视评估坏死。在第14天，EcSOD治疗组与对照组缺血肌肉中EcSOD表达上调2倍。处理的肌肉中的毛细血管密度（1.65±0.02毛细血管/纤维）比对照肌肉（0.78±0.17毛细血管/纤维，p<0.05）高1.9倍。缺血后第14天，EcSOD小鼠的灌注率恢复是对照小鼠的1.5倍（p<0.05）。EcSOD处理组小鼠的凋亡细胞核百分比为1.3 ± 0.4%，而对照组为4.2± 0.2%（p<0.001）。与对照小鼠相比，EcSOD中的肢体坏死也显著较低。AAV 9介导的骨骼肌中EcSOD的过表达显著改善了小鼠后肢缺血的恢复，这与治疗小鼠中改善的毛细血管密度和灌注比一致。
Neovascularization is a physiological repair process that is partly dependent on nitric oxide. Extracellular superoxide dismutase (EcSOD) is the major scavenger of superoxide and thus is an important regulator of nitric oxide bioavailability and thus protects against vascular dysfunction. We hypothesized that overexpression of EcSOD in skeletal muscle would improve recovery from hind-limb ischemia. Adeno associated virus (AAV) vectors expressing EcSOD or luciferase (control) from the Cytomegalovirus (CMV) promoter were cross-packaged into AAV9 capsids and injected IM into hind-limb muscles (1×1011 viral genomes(vg)/limb) of 12 wk-old mice. Ischemia was then induced after IM injections. Limb perfusion was serially measured by laser Doppler on days 0, 7 & 14 post-injection and values were expressed as a ratio relative to the non-ischemic limb. EcSOD expression was measured by Western blotting. Capillary density was documented by immunohistochemical staining for platelet endothelial cell adhesion molecule (PECAM). Apoptosis was assessed by Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and necrosis was visually evaluated daily. EcSOD expression was 2-fold up-regulated in EcSOD treated vs. control ischemic muscles at day 14. Capillary density was 1.9-fold higher in treated (1.65±0.02 capillaries/fiber) vs. control muscle (0.78±0.17 capillaries/fiber, p<0.05). Recovery of perfusion ratio at day 14 post-ischemia was 1.5-fold greater in EcSOD vs. control mice (p<0.05). The percentage of apoptotic nuclei was 1.3 ± 0.4% in EcSOD treated mice as compared to 4.2± 0.2% in controls (p<0.001). Limb necrosis was also significantly lower in EcSOD vs. control mice. AAV9-mediated overexpression of EcSOD in skeletal muscle significantly improves recovery from hind-limb ischemia in mice, consistent with improved capillary density and perfusion ratios in treated mice.
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影响因子：
30.8
作者：
KAPLITT, MG;LEONE, P;DURING, MJ
通讯作者：
DURING, MJ
影响因子：
14
作者：
Layman, Hans;Spiga, Maria-Grazia;Andreopoulos, Fotios M.
通讯作者：
Andreopoulos, Fotios M.
影响因子：
12.4
作者：
Prasad, Konkal-Matt R.;Xu, Yaqin;French, Brent A.
通讯作者：
French, Brent A.
影响因子：
3.7
作者：
Oshikawa J;Urao N;Kim HW;Kaplan N;Razvi M;McKinney R;Poole LB;Fukai T;Ushio-Fukai M
通讯作者：
Ushio-Fukai M
DOI：
10.1016/j.bbrc.2003.08.134
发表时间：
2003-10-10
影响因子：
3.1
作者：
Fukino, K;Sata, M;Nagai, R
通讯作者：
Nagai, R