Pluronic L64-mediated stable HIF-1α expression in muscle for therapeutic angiogenesis in mouse hindlimb ischemia.
Pluronic L64-mediated stable HIF-1α expression in muscle for therapeutic angiogenesis in mouse hindlimb ischemia.
复制标题
Pluronic (R) L64 介导的肌肉中稳定的 HIF-1 α 表达用于治疗小鼠后肢缺血的血管生成
DOI：
10.2147/ijn.s65353
复制
发表时间：
2014
影响因子：
8
通讯作者：
Gu Z
中科院分区：
文献类型：
--
作者：
Song H;Liu S;Li C;Geng Y;Wang G;Gu Z
肌内注射质粒 DNA (pDNA) 来表达治疗蛋白是治疗许多疾病的一种有前途的方法。然而，治疗应用通常受到基因传递效率和表达水平的阻碍。在本研究中，对基于pDNA的基因治疗系统中的关键因素（例如基因递送材料、治疗基因及其调控元件）进行了优化，以建立治疗小鼠后肢缺血的集成系统。结果表明，Pluronic® L64 (L64) 是一种高效、安全的基因导入小鼠骨骼肌的材料。它还显示出以浓度依赖性方式促进体内血管生成的内在能力，这可能是通过激活活化 B 细胞 (NF-κB) 调节的血管生成因子的核因子 kappa 轻链增强子来实现的。 0.1% L64 与杂合基因启动子 (pSC) 的组合增加了基因表达水平，延长了基因表达持续时间，并增加了转染肌纤维的数量。在小鼠缺血肢体中，由L64和基于pSC的编码缺氧诱导因子1-α三突变体（HIF-1αtri）的表达质粒组成的基因药物（pSC-HIF1αtri/L64）提高了稳定HIF-1α的表达，进而提高了多种血管生成因子的表达。结果，缺血肢体的功能恢复加速，足部坏死减少，血液再灌注加快，毛细血管密度更高。这些结果表明pSC-HIF1αtri/L64组合为治疗周围血管疾病，特别是严重的肢体缺血提供了潜在且方便的途径。
Intramuscular injection of plasmid DNA (pDNA) to express a therapeutic protein is a promising method for the treatment of many diseases. However, the therapeutic applications are usually hindered by gene delivery efficiency and expression level. In this study, critical factors in a pDNA-based gene therapy system, such as gene delivery materials, a therapeutic gene, and its regulatory elements, were optimized to establish an integrated system for the treatment of mouse hindlimb ischemia. The results showed that Pluronic® L64 (L64) was an efficient and safe material for gene delivery into mouse skeletal muscle. It also showed intrinsic ability to promote in vivo angiogenesis in a concentration-dependent manner, which might be through the activation of nuclear factor kappa-light-chain-enhancer of activated B cell (NF-κB)-regulated angiogenic factors. The combination of 0.1% L64 with a hybrid gene promoter (pSC) increased the gene expression level, elongated the gene expression duration, and enhanced the number of transfected muscle fibers. In mice ischemic limbs, a gene medicine (pSC-HIF1αtri/L64) composed of L64 and pSC-based expression plasmid encoding hypoxia-inducible factor 1-alpha triple mutant (HIF-1αtri), improved the expression of stable HIF-1α, and in turn, the expression of multiple angiogenic factors. As a result, the ischemic limbs showed accelerated function recovery, reduced foot necrosis, faster blood reperfusion, and higher capillary density. These results indicated that the pSC-HIF1αtri/L64 combination presented a potential and convenient venue for the treatment of peripheral vascular diseases, especially critical limb ischemia.