Key Laboratory of Polymer Ecomaterials, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China; Department of Microbiology and Molecular Genetics, University of the Punjab, 54590 Lahore, Pakistan.
Key Laboratory of Polymer Ecomaterials, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China.
Acta Biomater. 2017 May;54:58-68. doi: 10.1016/j.actbio.2016.12.048. Epub 2016 Dec 24.
Parkinson's Disease (PD) is a chronic neurodegenerative disorder characterized by motor deficits which result from the progressive loss of dopaminergic neurons. Gene therapy using growth factors such as VEGF seems to be a viable approach for potential therapeutic treatment of PD. In this study, we utilized a novel non-viral gene carrier designated as PEI-PLL synthesized by our laboratory to deliver VEGF gene to study its effect by using both cell culture as well as animal models of PD. For cell culture experiments, we utilized 6-hydroxydopamine (6-OHDA) mediated cell death model of MN9D cells following transfection with either a control plasmid or VEGF expressing plasmid. As compared to control transfected cells, PEI-PLL mediated VEGF gene delivery to MN9D cells resulted in increased cell viability, increase in the number of Tyrosine hydroxylase (TH) positive cells and decreased apoptosis following 6-OHDA insult. Next, we studied the therapeutic potential of PEI-PLL mediated VEGF gene delivery in SNPc by using unilateral 6-OHDA Medial forebrain bundle (MFB) lesion model of PD in rats. VEGF administration prevented the loss of motor functions induced by 6-OHDA as determined by behavior analysis. Similarly, VEGF inhibited the 6-OHDA mediated loss of DA neurons in Substantia Nigra Pars Compacta (SNPc) as well as DA nerve fibers in striatum as determined by TH immunostaining. In addition, PEI-PLL mediated VEGF gene delivery also prevented apoptosis and microglial activation in PD rat models. Together, these results clearly demonstrated the beneficial effects of PEI-PLL mediated VEGF gene delivery on dopaminergic system in both cell culture and animal models of PD.
In this report, we exploited the potential of PEI-PLL to deliver VEGF gene for the potential therapeutic treatment of PD by using both cell culture and animal models of PD. To the best of our knowledge, this is the first report describing the use of novel polymeric gene carriers for the delivery of VEGF gene to DA neurons with improved transfection efficiency. Finally, the study will lead to a significant advancement in the field of non-viral PD gene therapy treatment.
帕金森病（PD）是一种慢性神经退行性疾病，其运动缺陷是由于多巴胺能神经元的进行性丧失引起的。使用生长因子（如 VEGF）的基因治疗似乎是治疗 PD 的一种可行方法。在这项研究中，我们利用了我们实验室合成的一种新型非病毒基因载体，即 PEI-PLL，将 VEGF 基因传递到细胞培养和 PD 动物模型中，以研究其效果。对于细胞培养实验，我们利用 6-羟多巴胺（6-OHDA）介导的 MN9D 细胞死亡模型，转染对照质粒或表达 VEGF 的质粒。与对照转染细胞相比，PEI-PLL 介导的 VEGF 基因传递到 MN9D 细胞可增加细胞活力，增加酪氨酸羟化酶（TH）阳性细胞数量，并减少 6-OHDA 损伤后的细胞凋亡。接下来，我们在大鼠 PD 的单侧 6-OHDA 中脑束（MFB）损伤模型中研究了 SNPc 中 PEI-PLL 介导的 VEGF 基因传递的治疗潜力。VEGF 给药可防止行为分析确定的 6-OHDA 诱导的运动功能丧失。同样，VEGF 抑制了 TH 免疫染色确定的 SNpc 中 DA 神经元和纹状体中 DA 神经纤维的 6-OHDA 介导的丧失。此外，PEI-PLL 介导的 VEGF 基因传递还可防止 PD 大鼠模型中的细胞凋亡和小胶质细胞激活。综上所述，这些结果清楚地表明了 PEI-PLL 介导的 VEGF 基因传递对细胞培养和 PD 动物模型中多巴胺能系统的有益影响。
在本报告中，我们利用了 PEI-PLL 的潜力，通过使用 PD 的细胞培养和动物模型，通过细胞培养和动物模型，将 VEGF 基因传递到细胞培养和动物模型中，用于 PD 的潜在治疗。据我们所知，这是第一篇描述使用新型聚合物基因载体将 VEGF 基因递送到 DA 神经元以提高转染效率的研究报告。最后，这项研究将推动非病毒 PD 基因治疗治疗领域的重大进展。
