Soleimani Mitra, Ghasemi Nazem, Chamnari Fatemeh Mohammadi
1Department of Anatomical Science and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
2Faculty of Paramedicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Comp Clin Path. 2018;27(4):1023-1028. doi: 10.1007/s00580-018-2696-3. Epub 2018 Mar 19.
Parkinson's disease (PD) is one of the most neurodegenerative disorders which can lead to severe neural disability and neurological defects. Cell-based therapy using fully differentiated cells is a new method for the treatment of this abnormal condition. In the present study, we investigated the effects of 6-bromoindirubin-3'-oxime (BIO) on dopaminergic differentiation of human immortalized RenVm cells in order to obtain a set of fully differentiated cells for transplantation in Parkinson's disease. To this end, the immortalized RenVm cells were induced to dopaminergic differentiation using a neuro basal medium supplemented with N2 and different concentrations (75, 150, 300, 600, and 1200 nM) of BIO for 4, 8, and 12 days. The efficiency of dopaminergic differentiation was determined using immunocytochemistry for tyrosine hydroxylase expressions. In addition, the expression of a β-catenin marker was measured using the western blot technique. The results of immunocytochemistry revealed that the mean percentage of Tuj1- and TH-positive sells in 150- and 300-nM-BIO-treated groups was significantly increased compared to that of other groups ( ≤ 0.01). In addition, the expression of the β-catenin marker was higher in these groups as compared with that of other groups. Overall, BIO through its effect on the Wnt-Frizzled signaling pathway can promote dopaminergic differentiation of RenVm cells in a dose-dependent manner.
帕金森病（PD）是最常见的神经退行性疾病之一，可导致严重的神经功能残疾和神经缺陷。使用完全分化细胞的细胞疗法是治疗这种异常病症的一种新方法。在本研究中，我们研究了6-溴靛玉红-3'-肟（BIO）对人永生化RenVm细胞多巴胺能分化的影响，以便获得一组用于帕金森病移植的完全分化细胞。为此，使用补充有N2和不同浓度（75、150、300、600和1200 nM）BIO的神经基础培养基诱导永生化RenVm细胞进行多巴胺能分化4、8和12天。使用免疫细胞化学检测酪氨酸羟化酶表达来确定多巴胺能分化的效率。此外，使用蛋白质印迹技术测量β-连环蛋白标志物的表达。免疫细胞化学结果显示，与其他组相比，150 nM和300 nM BIO处理组中Tuj1和TH阳性细胞的平均百分比显著增加（≤0.01）。此外，与其他组相比，这些组中β-连环蛋白标志物的表达更高。总体而言，BIO通过其对Wnt-Frizzled信号通路的作用，可以剂量依赖性方式促进RenVm细胞的多巴胺能分化。
