Intraventricular Administration Of Bmp-7 In Stroke Anima

中风和相关血管事件可导致中枢神经系统（CNS）的广泛损伤，并导致严重的运动和认知障碍。已经表明，营养因子治疗可以减少损伤的程度并在损伤后恢复受损的神经元。我们先前已经测试了骨形态发生蛋白-7（BMP-7），生长因子的转化生长因子-B超家族的成员，在局灶性脑缺血损伤中的作用。我们发现BMP-7在脑室内给药后具有深刻的神经保护作用。我们现在已经着手测试其在缺血脑中的分布，以确定BMP-7通过脑实质扩散的速度。我们还确定了它是否对损伤后短期神经元中中间丝的表达有影响，作为其恢复活性的部分基础。最后，我们研究了中风后对运动行为的影响。将成年雄性Sprague-Dawley大鼠的右大脑中动脉闭塞60分钟，24小时后，将BMP-7或载体以25 μ l（1 μ g/μ l）的剂量注射到同侧的侧脑室中。在BMP-7给药后1、3、6和24小时处死动物。在脑实质中的定位方面，我们发现BMP-7在注射后1小时主要存在于室管膜中，而在注射后3小时它延伸到一些邻近的神经元。然而，6小时和24小时组的神经元，主要是在纹状体和丘脑有显着的BMP-7免疫反应标记。在接受载体注射的动物中，在注射侧神经丝表达似乎相当弱，而在注射BMP-7后6小时和24小时观察到神经丝染色的显著上调。在24小时观察到显著的神经突生长，具有明显的生长锥，特别是在腹侧纹状体和腹侧纹状体。这些结果表明，BMP-7注射可能有助于表达的中间神经元丝缺血性损伤后，此外，注射的因子可以成为纳入神经元位于几毫米的侧脑室。 用水合氯醛麻醉成年Sprague-Dawley大鼠。大脑中动脉（MCA）被通过右侧颈内动脉插入的细丝短暂闭塞。在60分钟缺血后移除细丝以允许再灌注。在MCA闭塞（MCAo）后24小时处死一些动物以检测BMP-7 mRNA表达。其他动物在MCA闭塞后24小时接受单剂量的静脉内BMP-7或载体，并用于随后的行为研究和BMP-7免疫染色。在未处理的动物中，MCAo后24小时BMP-7 mRNA上调。BMP-7免疫反应性在接受BMP-7静脉注射的动物中，在MCAo后第6天在缺血侧海马/齿状回上呈剂量依赖性增加。接受BMP-7的动物也显示出从第7天到第14天身体不对称性的降低和在MCAo后第14天运动活性的增加。 我们的数据表明，BMP-7，给予胃肠外中风后，可以通过缺血侧的血脑屏障，并诱导中风动物在较长的测试时间的行为恢复。

Stroke and related vascular events can cause widespread damage to the central nervous system (CNS) and result in significant motor and cognitive impairments. It has been suggested that trophic factor treatment may reduce the extent of damage and restore damaged neurons following the injury. We have previously tested the effects of bone morphogenetic protein-7 (BMP-7), a member of the transforming growth factor-B superfamily of growth factors, in focal brain ischemic injury. We found BMP-7 to have profound neuroprotective effects after intraventricular administration. We have now proceeded to test its distribution in the ischemic brain, in order to determine how rapidly BMP-7 may diffuse through brain parenchyma. We also determined whether it has effects on expression of intermediate filaments in neurons short-term after the injury as a partial basis for its restorative activity. Finally we examined effects on motor behavior after stroke. Adult male Sprague-Dawley rats were subjected to a 60-minute occlusion of the right middle cerebral artery, and 24 hours thereafter, injections of BMP-7 or vehicle into the lateral ventricle on the ipsilateral side, at a dose of 25 ul (1 ug/ul), were performed. The animals were sacrificed 1, 3, 6, and 24 hours following the BMP-7 administration. In terms of localization in the brain parenchyma, we found that BMP-7 was mostly present in the ependyma 1 hour following injection, while it extended to a few adjacent neurons at 3 hours post-injection. However, the 6- and 24-hour groups both had significant BMP-7-immunoreactive labeling in neurons, primarily in the striatum and thalamus. Neurofilament expression appeared to be fairly weak on the injected side in animals that had received the vehicle injection, whereas a significant upregulation in neurofilament staining was observed at 6- and 24 hours post-injection of BMP-7. Significant neurite outgrowth, with obvious growth cones, was observed at 24 hours, particularly in the ventral striatum and accumbens. These results suggest that BMP-7 injection may aid in the expression of intermediate neuronal filaments following an ischemic injury, and furthermore that the injected factor can become incorporated into neurons located several millimeters from the lateral ventricle. Adult Sprague-Dawley rats were anesthetized with chloral hydrate. The middle cerebral artery (MCA) was transiently occluded by a filament, inserted through the right internal carotid artery. The filament was removed after 60-min ischemia to allow reperfusion. Some animals were killed 24 hours after MCA occlusion (MCAo) to examine BMP-7 mRNA expression. Other animals received a single dose of intravenous BMP-7 or vehicle at 24 hours after MCA occlusion and were used for subsequent behavioral studies and BMP-7 immunostaining. BMP-7 mRNA was upregulated 24 hours after MCAo in non-treated animals. BMP-7-immunoreactivity was dose-dependently increased on the ischemic side hippocampus/dentate on the 6th day after MCAo in animals receiving intravenous injection of BMP-7. Animals receiving BMP-7 also showed a decrease in body asymmetry from days 7 to day 14 and an increase in locomotor activity on day 14 after MCAo. Our data indicate that BMP-7, given parenterally after stroke, can pass through the blood brain barrier on the ischemic side and induce behavioral recovery in stroke animals at longer testing times.