Division of Neurosurgery, Department of Surgery, The Chinese University of Hong Kong, Shatin, Hong Kong.
Neurosurgery. 2010 Aug;67(2):357-65; discussion 365-6. doi: 10.1227/01.NEU.0000371983.06278.B3.
Spontaneous intracerebral hemorrhage (ICH) carries a high mortality rate, with survivors commonly left with permanent neurological deficits. Mesenchymal stem cell (MSC) transplantation promotes functional recovery in experimental ICH, and treatment with hepatocyte growth factor (HGF) is beneficial in ischemic stroke.
We hypothesize that transplantation of MSCs with previous transduction of HGF has an additive effect in promoting neurological recovery through myelin and axonal regeneration.
HGF transduction to human umbilical cord-derived MSCs using lentiviral plasmid pWPI-HGF-GFP was prepared. One week after a collagenase-induced ICH, 80 male Sprague-Dawley rats were divided into 3 groups for stereotactic injection of phosphate-buffered saline (group I), MSC transplant (group II), and HGF-transduced MSC transplant (group III), respectively, into the left ventricle. The animals were assessed weekly for 5 weeks using the Rotarod motor function test, at which time they were killed for Luxol fast blue myelin staining and appropriate immunohistochemistry and Western blotting.
Animals receiving transplanted HGF-transduced MSCs (group III) exhibited significantly better motor function recovery than animals treated with MSCs alone (group II), which in turn performed better than the phosphate-buffered saline controls at 2 weeks after transplantation. Luxol fast blue staining of myelin displayed significantly less demyelination and significantly higher reactivity in myelin basic protein and growth-associated protein-43 in immunohistochemistry and Western blotting and significantly reduced myelin-associated glycoprotein activity in group III animals.
Animals transplanted with HGF-transduced MSCs 1 week after experimental ICH were shown to achieve a better neurological recovery. This improved neurological recovery from ICH is attributed to nerve fiber remyelination and axonal regeneration.
自发性脑出血（ICH）死亡率高，幸存者常遗留永久性神经功能缺损。间充质干细胞（MSC）移植可促进实验性 ICH 后的功能恢复，肝细胞生长因子（HGF）治疗有益于缺血性脑卒中。
我们假设，先前转导 HGF 的 MSC 移植通过髓鞘和轴突再生具有促进神经恢复的附加作用。
使用慢病毒质粒 pWPI-HGF-GFP 制备 HGF 转导到人脐带来源的 MSC。胶原酶诱导 ICH 后 1 周，80 只雄性 Sprague-Dawley 大鼠分为 3 组，分别立体定向注射磷酸盐缓冲液（I 组）、MSC 移植（II 组）和 HGF 转导的 MSC 移植（III 组）到左心室。动物每周通过转棒运动功能测试进行评估，持续 5 周，此时处死动物进行卢索快速蓝髓鞘染色和适当的免疫组织化学和 Western blot。
接受移植的 HGF 转导 MSC（III 组）的动物表现出明显更好的运动功能恢复，优于单独接受 MSC 治疗的动物（II 组），后者在移植后 2 周时比磷酸盐缓冲液对照组表现出更好的运动功能恢复。免疫组织化学和 Western blot 中的髓鞘碱性蛋白和生长相关蛋白-43的髓鞘脱失明显减少，髓鞘相关糖蛋白活性明显降低，卢索快速蓝染色显示髓鞘脱失明显减少。
ICH 后 1 周接受 HGF 转导 MSC 移植的动物表现出更好的神经恢复。这种改善的 ICH 神经恢复归因于神经纤维再髓鞘和轴突再生。
