1. Department of Biology, School of Biology, Damghan University, Damghan, Iran ; 2.Institute of Biological Sciences, Damghan University, Damghan, Iran.
Cell J. 2013 Spring;15(1):55-64. Epub 2013 May 5.
There is longstanding experimental and clinical evidence that supports the idea that replacement of dopaminergic (DAergic) neurons can ameliorate functional disabilities of Parkinson's disease (PD). The purpose of the present study is to examine the efficacy of transplantation of rat bone marrow stromal cell (BMSCs)-derived tyrosine hydroxylase-positive (TH(+)) cells induced by deprenyl into 6-hydroxydopamine (6-OHDA)-lesioned rat models, using behavioral tests and immunohistochemical evaluations.
In this experimental study, undifferentiated BrdU-labeled BMSCs were incubated in serum-free medium that contained 10(-8) M deprenyl for 24 hours. Afterwards, BMSCs were cultured for 48 hours in α-minimal essential medium (α-MEM) supplemented with 10% FBS, then differentiated into TH(+) neurons. We randomly divided 24 hemiparkinsonian rats as follows: group 1 (control) received only medium, while groups 2 and 3 were injected with 2×10(5) BMSCs and deprenyl-treated cells in 4 µl medium. Injections were made into the injured strata of the rats. Rotational behavior in response to apomorphine was tested before transplantation and at 2, 4, and 6 weeks post-graft. Animals were then sacrificed, and the brains were extracted for immunohistochemical and electron microscopic studies.
Apomorphine-induced rotation analysis indicated that animals with grafted cells in groups 2 and 3 exhibited significantly less rotational behavior than those in the control group at 2, 4, and 6 weeks after transplantation. Immunohistochemical analysis demonstrated that BrdU-labeled cells expressed specific neuronal markers, such as NF 200 and TH, at the implantation site. The presence of TH(+) cells in conjunction with the reduction in rotation might show the capacity of grafted cells to release dopamine. Ultrastructural analysis revealed the presence of immature neurons and astrocyte-like cells at the graft site.
TH(+) neurons induced by deprenyl can be considered as a cell source for PD autograft therapy.
长期的实验和临床证据支持这样一种观点，即多巴胺能（DAergic）神经元的替代可以改善帕金森病（PD）的功能障碍。本研究的目的是通过行为测试和免疫组织化学评估，检查去甲丙咪嗪诱导的大鼠骨髓基质细胞（BMSCs）衍生的酪氨酸羟化酶阳性（TH（+））细胞移植到 6-羟多巴胺（6-OHDA）损伤大鼠模型中的疗效。
在这项实验研究中，未分化的 BrdU 标记的 BMSCs 在无血清培养基中孵育，其中包含 10（-8）M 去甲丙咪嗪 24 小时。之后，BMSCs 在含 10％FBS 的α最小必需培养基（α-MEM）中培养 48 小时，然后分化为 TH（+）神经元。我们将 24 只半帕金森病大鼠随机分为以下三组：第 1 组（对照组）仅接受培养基，而第 2 组和第 3 组分别接受 2×10（5）个 BMSCs 和去甲丙咪嗪处理的细胞，剂量为 4μl 培养基。注射到大鼠损伤的脑区。在移植前和移植后 2、4 和 6 周，通过阿朴吗啡测试旋转行为。然后处死动物，提取大脑进行免疫组织化学和电子显微镜研究。
阿朴吗啡诱导的旋转分析表明，与对照组相比，移植后 2、4 和 6 周，移植细胞的第 2 组和第 3 组动物的旋转行为明显减少。免疫组织化学分析表明，在植入部位，BrdU 标记的细胞表达特定的神经元标志物，如 NF 200 和 TH。存在 TH（+）细胞与旋转减少可能表明移植细胞释放多巴胺的能力。超微结构分析显示在移植物部位存在未成熟神经元和星形胶质样细胞。
去甲丙咪嗪诱导的 TH（+）神经元可被视为 PD 自体移植治疗的细胞来源。
