Antioxidant enzyme gene delivery to protect from HIV-1 gp120-induced neuronal apoptosis
Antioxidant enzyme gene delivery to protect from HIV-1 gp120-induced neuronal apoptosis
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DOI：
10.1038/sj.gt.3302821
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发表时间：
2006-12-01
期刊：
影响因子：
5.1
通讯作者：
Strayer, D. S.
中科院分区：
文献类型：
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作者：
Agrawal, L.;Louboutin, J-P;Strayer, D. S.
人类免疫缺陷病毒-1(HIV-1)感染中枢神经系统(CNS)可导致神经元丢失和中枢神经系统功能的进行性恶化：HIV-1基因产物，尤其是gp120，可诱导自由基介导的细胞凋亡。活性氧物种(ROS)是这些效应的潜在介体。神经元在暴露于gp120后容易形成ROS，因此可以通过抗氧化酶如铜/锌超氧化物歧化酶(SOD1)和/或谷胱甘肽过氧化物酶(GPX1)来保护神经元免受ROS介导的损伤。这两种酶都能清除氧自由基。由于它们是高效的神经元基因传递工具，重组SV40衍生载体被用于这些研究。用携带人SOD1和/或GPX1基因的rSV40载体转导培养的NT2细胞成熟神经元和原代胎儿神经元，然后暴露于gp120。用末端脱氧核苷酸转移酶介导的缺口末端标记法检测细胞凋亡率。免疫染色检测，两组神经元的转导效率均为95%。用Western blotting和酶活性直接测定法确定转基因的表达。Gp120诱导高比例无保护的NT2-N细胞发生凋亡。在gp120攻击前用SV(SOD1)和SV(GPX1)转导可使神经细胞凋亡减少90%。在依次使用两种载体处理的细胞中，甚至可以看到更大的保护作用。单独给予或联合给予，它们可以保护神经细胞免受HIV1-gp120诱导的细胞凋亡。我们在体内测试了rSV40 S是否能将抗氧化酶输送到中枢神经系统：脑内注射SV(SO1)或SV(GPX1)到大鼠脑尾壳核可获得良好的转基因神经元表达。在体内，使用SV(SOD1)的转导也可以保护gp120诱导的神经元在两者注射到大鼠尾壳核后诱导的细胞凋亡。因此，SOD1和GPX1可以通过SV40载体在体外或体内传递。这种方法可能值得考虑用于HIV-1引起的脑病的治疗。
Human immunodeficiency virus-1 (HIV-1) infection in the central nervous system (CNS)may lead to neuronal loss and progressively deteriorating CNS function: HIV-1 gene products, especially gp120, induce free radical-mediated apoptosis. Reactive oxygen species (ROS), are among the potential mediators of these effects. Neurons readily form ROS after gp120 exposure, and so might be protected from ROS-mediated injury by antioxidant enzymes such as Cu/Zn-superoxide dismutase (SOD1) and/or glutathione peroxidase (GPx1). Both enzymes detoxify oxygen free radicals. As they are highly efficient gene delivery vehicles for neurons, recombinant SV40-derived vectors were used for these studies. Cultured mature neurons derived from NT2 cells and primary fetal neurons were transduced with rSV40 vectors carrying human SOD1 and/or GPx1 cDNAs, then exposed to gp120. Apoptosis was measured by terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay. Transduction efficiency of both neuron populations was > 95%, as assayed by immunostaining. Transgene expression was also ascertained by Western blotting and direct assays of enzyme activity. Gp120 induced apoptosis in a high percentage of unprotected NT2-N. Transduction with SV(SOD1) and SV(GPx1) before gp120 challenge reduced neuronal apoptosis by > 90%. Even greater protection was seen in cells treated with both vectors in sequence. Given singly or in combination, they protect neuronal cells from HIV 1-gp120 induced apoptosis. We tested whether rSV40 s can deliver antioxidant enzymes to the CNS in vivo: intracerebral injection of SV(SOD1) or SV(GPx1) into the caudate putamen of rat brain yielded excellent transgene expression in neurons. In vivo transduction using SV(SOD1) also protected neurons from subsequent gp120-induced apoptosis after injection of both into the caudate putamen of rat brain. Thus, SOD1 and GPx1 can be delivered by SV40 vectors in vitro or in vivo. This approach may merit consideration for therapies in HIV-1-induced encephalopathy.