Marzban Mohsen, Mousavizadeh Kazem, Bakhshayesh Masoomeh, Vousooghi Nasim, Vakilzadeh Gelareh, Torkaman-Boutorabi Anahita
Department of Neuroscience, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Cellular and Molecular Research Center and Department of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran.
Iran Biomed J. 2018 Sep;22(5):312-21. doi: 10.29252/ibj.22.5.312. Epub 2018 Feb 7.
Bone marrow mesenchymal stem cells (BM-MSCs) elicit neuroprotective effects, and their repair ability has been investigated in different experimental models. We aimed to investigate the effect of multiple i.p. BM-MSCs injections in the cuprizone model of multiple sclerosis in mice.
Adult male C57BL/6 mice (n = 40) were fed a regular diet or a diet containing cuprizone (0.2% w/w) for 6 six weeks. Bone marrow samples were taken from patients with spinal cord injury. BM-MSCs (2 × 106 in 1 milliliter medium) were administered intraperitoneally for two consecutive weeks at the end of the forth weeks of cuprizone administration. Animals (n = 12) were perfused with 10% paraformaldehyde at the end of sixth week. The brains were sectioned coronally in 6-8-μm thickness (-2.3 to 1.8 mm from bregma). The sections were stained by luxol fast blue-cresyl violet, and images were captured via a microscope. Demyelination ratio was estimated in corpus callosum in a blind manner. A quantitative real-time PCR was used to measure the myelin basic protein gene expression at sixth week.
Histologically, cuprizone induced demyelination in the corpus callosum. Demyelinated area was diminished in the corpus callosum of cell-administered group. Cuprizone could decrease myelin-binding protein mRNAs expression in corpus callosum, which was significantly recovered after BM-MSCs injections.
Our data indicated a remyelination potency of multiple i.p. BM-MSCs in the cuprizone model of multiple sclerosis in mice.
骨髓间充质干细胞(BM-MSCs)具有神经保护作用,其修复能力已在不同实验模型中得到研究。我们旨在研究多次腹腔注射BM-MSCs对小鼠铜螯合剂诱导的多发性硬化模型的影响。
将40只成年雄性C57BL/6小鼠分为两组,一组给予常规饮食,另一组给予含铜螯合剂(0.2% w/w)的饮食,持续6周。从脊髓损伤患者处采集骨髓样本。在给予铜螯合剂4周结束时,连续两周腹腔注射BM-MSCs(2×10⁶个细胞/1毫升培养基)。在第6周结束时,对12只动物用10%多聚甲醛进行灌注固定。将大脑冠状切成6 - 8μm厚的切片(距前囟-2.3至1.8 mm)。切片用Luxol快速蓝-甲酚紫染色,通过显微镜采集图像。以盲法估计胼胝体的脱髓鞘比例。在第6周时,采用定量实时PCR检测髓鞘碱性蛋白基因表达。
组织学上,铜螯合剂诱导胼胝体脱髓鞘。细胞给药组胼胝体的脱髓鞘区域减少。铜螯合剂可降低胼胝体中髓鞘结合蛋白mRNA的表达,BM-MSCs注射后该表达显著恢复。
我们的数据表明,多次腹腔注射BM-MSCs对小鼠铜螯合剂诱导的多发性硬化模型具有髓鞘再生能力。