Department of Anatomy and Neurobiology, Tongji University School of Medicine, and Department of Plastic and Reconstructive Surgery, 9th People's Hospital, Shanghai, China.
Neurosurgery. 2011 Mar;68(3):691-704. doi: 10.1227/NEU.0b013e3182098a8a.
Intracerebral hemorrhage (ICH) represents at least 15% of all strokes in the Western population and a considerably higher proportion at 50% to 60% in the Oriental population.
To investigate whether administration of bone marrow stem cells (BMSCs) overexpressing glial cell line-derived neurotrophic factor (GDNF) provides more efficient neuroprotection for rats with ICH and neurons exposed to hypoxia/reoxygenation.
Primary rat BMSCs were transfected with rat GDNF gene using virus vector (GDNF/BMSCs) and blank virus plasmid (BVP/BMSCs). Primary rat cortical neurons of rats were exposed to hypoxia and then reoxygenated with GDNF/BMSCs (GDNF/BMSCs group) or BVP/BMSCs (BMSCs group) treatment for 12 hours and 1, 2, 3, and 5 days. Hoechst 33258 staining was used to evaluate apoptosis. GDNF/BMSCs, BVP/BMSCs, and saline (GDNF/BMSCs, BMSCs, and control groups) were injected into the right striatum 3 days after rat ICH induced by injecting collagenase. Modified neurological severity scores and hematoxylin and eosin staining were performed to evaluate neurological function and lesion volume at 1 and 2 weeks after transplantation. Immunostaining was used to observe differentiation of grafted cells (neurofilament-200 for neurons, glial fibrillary acidic protein for astrocytes). The GDNF level and apoptosis were evaluated by Western blotting and terminal deoxynucleotidyl transferase dUTP nick-end labeling, respectively.
The GDNF/BMSCs group had significantly lowered apoptosis compared with the BMSCs group at the given time. The GDNF/BMSCs group had significantly improved functional deficits and reduced lesion volume compared with the BMSCs group. Stable GDNF expression in the GDNF/BMSCs group was detected at the given time in the host brain. The neurofilament-positive grafted cells in the GDNF/BMSCs group were more numerous than in the BMSCs group. The GDNF/BMSCs group had significantly decreased apoptotic cells compared with the BMSCs group.
These results suggest that GDNF/BMSCs provide better neuroprotection for rats with ICH and neurons exposed to hypoxia/reoxygenation.
脑出血（ICH）占西方人群中风的 15%以上，而在东方人群中则高达 50%至 60%。
研究骨髓间充质干细胞（BMSCs）过表达胶质细胞源性神经营养因子（GDNF）对脑出血大鼠及缺氧/复氧神经元的神经保护作用是否更有效。
采用病毒载体（GDNF/BMSCs）转染大鼠 GDNF 基因，空白病毒质粒（BVP/BMSCs）转染大鼠 BMSCs，建立 GDNF/BMSCs 组和 BVP/BMSCs 组。原代大鼠皮质神经元缺氧/复氧后分别用 GDNF/BMSCs（GDNF/BMSCs 组）和 BVP/BMSCs（BMSCs 组）处理 12 小时和 1、2、3、5 天。Hoechst 33258 染色法检测细胞凋亡。脑出血大鼠造模后 3 天，右纹状体分别注射 GDNF/BMSCs、BVP/BMSCs、生理盐水（GDNF/BMSCs 组、BMSCs 组和对照组）。移植后 1 周和 2 周，进行改良神经功能缺损评分和苏木精-伊红染色，评价神经功能和损伤体积。免疫组化观察移植细胞分化（神经元：神经丝-200；星形胶质细胞：胶质纤维酸性蛋白）。Western blot 法检测 GDNF 水平，末端脱氧核苷酸转移酶 dUTP 缺口末端标记法检测细胞凋亡。
与 BMSCs 组相比，GDNF/BMSCs 组在特定时间的细胞凋亡明显减少。与 BMSCs 组相比，GDNF/BMSCs 组的功能缺陷明显改善，损伤体积减小。在宿主大脑中，GDNF/BMSCs 组在特定时间内检测到稳定的 GDNF 表达。GDNF/BMSCs 组的神经丝阳性移植细胞明显多于 BMSCs 组。与 BMSCs 组相比，GDNF/BMSCs 组的凋亡细胞明显减少。
这些结果表明，GDNF/BMSCs 对脑出血大鼠及缺氧/复氧神经元具有更好的神经保护作用。
