人β-NGF 基因修饰的骨髓间充质干细胞对帕金森病大鼠行为学的影响

摘要

目的：探讨脑内移植人β-神经生长因子（β-NGF）基因修饰的骨髓间充质干细胞（MSCs）对帕金森病（PD）模型大鼠行为学的影响。方法：将成功制备的PD大鼠模型按随机数字法分为β-NGF基因修饰的MSCs移植组（β-NGF-MSCs组）、MSCs移植组（MSCs组）、DMEM/F12培养液纹状体内注射组（DMEM/F12组）和非移植组（PD模型组）。细胞移植后，动态观察PD模型大鼠行为学变化及脑组织形态学的变化。结果：β-NGF基因修饰的MSCs脑内移植能有效改善PD模型大鼠的行为学表现。细胞移植术后第2周、第4周和第6周，β-NGF-MSCs组在阿朴吗啡诱发下30 min内的平均旋转圈数均明显低于MSCs组和PD模型组（P＜0．05）。 DMEM/F12组和PD模型组的旋转行为手术前后比较无明显变化（P＞0．05）。免疫组织化学染色结果显示，移植细胞在PD模型大鼠脑内均能存活，并能够与宿主组织产生整合，具有良好的组织相容性；宿主的脑组织结构无破坏，无胶质瘢痕形成。PD模型大鼠纹状体内移植β-NGF基因修饰的MSCs后，可持续稳定表达β-NGF，其对PD模型大鼠旋转行为的改善明显优于单纯应用MSCs移植治疗。结论：β-NGF基因修饰的MSCs脑内移植能明显改善PD模型大鼠行为学症状，具有潜在的临床应用价值。%AIM:To investigate the effect of human β-nerve growth factor (β-NGF) gene-modified bone mar-row-derived mesenchymal stem cells (MSCs) transplantation on the rotational behavior improvement in a rat model of Par -kinson disease (PD).METHODS:The rat model of PD was established successfully and the animals were divided into 4 groups:β-NGF-MSCs group (transplanted with 5 ×10 5β-NGF-engineered MSCs), MSCs group (transplanted with 5 ×10 5 MSCs), DMEM/F12 group (5 μL transplantation medium was injected in the right striatum of the rats ) and PD model group (without transplantation).The rotational scores were assessed 2 weeks, 4 weeks and 6 weeks after transplantation. At different time points after transplantation , the rats were tested for apomorphine (APO)-induced rotational behavior and the brains of the PD model rats were examined by fluorescence microscopy and immunohistochemical staining .RESULTS:Transplantation of human β-NGF gene-modified MSCs effectively improved the behavioral performance in the rats .At the 2nd, 4th and 6th weeks after cell transplantation, the rotational frequencies after injection of APO decreased significantly inβ-NGF-MSCs group compared with MSCs group and PD group (P<0.05).Both β-NGF gene-modified MSCs and MSCs survived in the brains of PD model rats , had good compatibility with the host cells , and showed no signs of destroying the host and the glial cicatrisation.Theβ-NGF gene-modified MSCs expressed β-NGF stablely in the brains of PD model rats , and showed obvious improvement of the rotational behavior in the PD model rats induced by APO compared with MSCs group.CONCLUSION:The behavior of the rats with PD is significantly improved by transplanting β-NGF-modified MSCs in right striatum, and β-NGF gene therapy has potential clinical value .