Oral tetrahydrobiopterin improves the beneficial effect of adenoviral-mediated eNOS gene transfer after induction of hindlimb ischemia.
Oral tetrahydrobiopterin improves the beneficial effect of adenoviral-mediated eNOS gene transfer after induction of hindlimb ischemia.
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DOI：
10.1038/mt.2010.109
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发表时间：
2010-08
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通讯作者：
Jinglian Yan;Guodong Tie;A. Hoffman;Yagai Yang;P. Nowicki;L. Messina
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作者：
Jinglian Yan;Guodong Tie;A. Hoffman;Yagai Yang;P. Nowicki;L. Messina
我们测试了这样的假设：口服补充内皮一氧化氮合酶 (eNOS) 辅因子四氢生物蝶呤 (BH4) 可提高缺血性大鼠后肢 eNOS 基因转移的治疗效果。 BH4或媒介物在诱导后肢缺血前1周开始，而含有牛eNOS cDNA (AdeNOS)的重组腺病毒或媒介物[磷酸盐缓冲盐水(PBS)]在诱导缺血后10天动脉内注入缺血后肢。与单独接受 AdeNOS 的大鼠相比，接受饮食 BH4 和 eNOS 基因转移联合治疗的大鼠（[eNOS，+BH4]组）在缺血腓肠肌中具有更高的 eNOS 表达、磷酸化 eNOS 表达（Ser1177）、Ca2+ 依赖性 NOS 活性以及亚硝酸盐 + 硝酸盐浓度。与单独接受 AdeNOS 的大鼠相比，[eNOS，+BH4] 组在缺血性腓肠肌中表现出较少的硝基酪氨酸和较高的还原型谷胱甘肽：氧化型谷胱甘肽 (GSH:GSSG) 比例。与单独接受 AdeNOS 的大鼠相比，[eNOS，+BH4] 组在缺血后肢具有更大的血流恢复和更高的毛细血管：肌细胞比率。最后，[eNOS,+BH4]组的后肢肌肉坏死少于单独给予AdeNOS的大鼠。我们得出的结论是，BH4 辅助饮食疗法可增加缺血性腓肠肌内 eNOS 基因转移的有益作用，一氧化氮 (NO) 产生增加、氧化应激减少、血流恢复增强和坏死减少就证明了这一点。
We tested the hypothesis that oral supplementation with the endothelial nitric oxide synthase (eNOS) cofactor tetrahydrobiopterin (BH4) improved the therapeutic efficacy of eNOS gene transfer in the ischemic rat hindlimb. BH4or vehicle were begun 1 week before induction of hindlimb ischemia, whereas recombinant adenovirus containing bovine eNOS cDNA (AdeNOS) or vehicle [phosphate-buffered saline (PBS)] was infused intra-arterially into the ischemic hindlimb 10 days after induction of ischemia. Rats receiving co-treatment with dietary BH4and eNOS gene transfer (the [eNOS, +BH4] group) had greater eNOS expression, phospho-eNOS expression (Ser1177), Ca2+-dependent NOS activity, and nitrite + nitrate concentrations in the ischemic gastrocnemius than did rats receiving AdeNOS alone. The [eNOS, +BH4] group demonstrated less nitrotyrosine and a higher ratio of reduced:oxidized glutathione (GSH:GSSG) in the ischemic gastrocnemius muscle than did rats receiving AdeNOS alone. The [eNOS, +BH4] group had greater flow recovery and a higher capillary:myocyte ratio in the ischemic hindlimb than did rats receiving AdeNOS alone. Finally, the [eNOS,+BH4] group had less necrosis of hindlimb muscles than rats given AdeNOS alone. We conclude that adjunctive dietary therapy with BH4increases the beneficial effects of eNOS gene transfer within the ischemic gastrocnemius muscle, as evidenced by increased nitric oxide (NO) production, diminished oxidative stress, enhanced flow recovery, and reduced necrosis.