Yoshimura Shin-Ichi, Morishita Ryuichi, Hayashi Katsuhiko, Kokuzawa Jouji, Aoki Motokuni, Matsumoto Kunio, Nakamura Toshikazu, Ogihara Toshio, Sakai Noboru, Kaneda Yasufumi
Department of Neurosurgery, Gifu University School of Medicine, Gifu, Japan.
Hypertension. 2002 May;39(5):1028-34. doi: 10.1161/01.hyp.0000017553.67732.e1.
Although cerebral hypoperfusion caused by cerebral occlusive disease leads to cerebral ischemic events, an effective treatment has not yet been established. Recently, a novel therapeutic strategy for ischemic disease using angiogenic growth factors to expedite and/or augment collateral artery development has been proposed. Therapeutic angiogenesis might be useful for the treatment of cerebral occlusive disease. Hepatocyte growth factor (HGF) is a potent angiogenic factor, in addition to vascular endothelial growth factor (VEGF), whereas in the nervous system HGF also acts as neurotrophic factor. Therefore, we hypothesized that gene transfer of these angiogenic growth factors could induce angiogenesis, thus providing an effective therapy for cerebral hypoperfusion or stroke. In this study, we employed a highly efficient gene transfer method, the viral envelop (Hemagglutinating Virus of Japan [HVJ]-liposome) method, because we previously documented that beta-galactosidase gene could be transfected into the brain by the HVJ-liposome method. Indeed, we confirmed wide distribution of transgene expression using beta-galactosidase via injection into the subarachnoid space. Of importance, transfection of HGF or VEGF gene into the subarachnoid space 7 days before occlusion induced angiogenesis on the brain surface as assessed by alkaline phosphatase staining (P<0.01). In addition, significant improvement of cerebral blood flow (CBF) was observed by laser Doppler imaging (LDI) 7 days after occlusion (P<0.01). Unexpectedly, transfection of HGF or VEGF gene into the subarachnoid space immediately after occlusion of the bilateral carotid arteries also induced angiogenesis on the brain surface and had a significant protective effect on the impairment of CBF by carotid occlusion (P<0.01). Interestingly, coinjection of recombinant HGF with HGF gene transfer revealed a further increase in CBF (P<0.01). Here, we demonstrated successful therapeutic angiogenesis using HGF or VEGF gene transfer into the subarachnoid space to improve cerebral hypoperfusion, thus providing a new therapeutic strategy for cerebral ischemic disease.
尽管由脑闭塞性疾病引起的脑灌注不足会导致脑缺血事件，但尚未确立有效的治疗方法。最近，有人提出了一种使用血管生成生长因子来加速和/或增强侧支动脉发育的缺血性疾病新治疗策略。治疗性血管生成可能对脑闭塞性疾病的治疗有用。肝细胞生长因子（HGF）是一种强大的血管生成因子，除血管内皮生长因子（VEGF）外，而在神经系统中HGF还作为神经营养因子发挥作用。因此，我们假设这些血管生成生长因子的基因转移可以诱导血管生成，从而为脑灌注不足或中风提供有效的治疗方法。在本研究中，我们采用了一种高效的基因转移方法，即病毒包膜（日本血凝病毒[HVJ]-脂质体）方法，因为我们之前记录了β-半乳糖苷酶基因可以通过HVJ-脂质体方法转染到大脑中。事实上，我们通过向蛛网膜下腔注射β-半乳糖苷酶证实了转基因表达的广泛分布。重要的是，在闭塞前7天将HGF或VEGF基因转染到蛛网膜下腔，通过碱性磷酸酶染色评估，诱导了脑表面的血管生成（P<0.01）。此外，在闭塞后7天通过激光多普勒成像（LDI）观察到脑血流量（CBF）有显著改善（P<0.01）。出乎意料的是，在双侧颈动脉闭塞后立即将HGF或VEGF基因转染到蛛网膜下腔也诱导了脑表面的血管生成，并对颈动脉闭塞引起的CBF损伤有显著的保护作用（P<0.01）。有趣的是，重组HGF与HGF基因转移共同注射显示CBF进一步增加（P<0.01）。在这里，我们证明了通过将HGF或VEGF基因转移到蛛网膜下腔成功进行治疗性血管生成以改善脑灌注不足，从而为脑缺血性疾病提供了一种新的治疗策略。
