多不饱和脂肪酸对人脐带间充质干细胞生物学特性的影响

发布时间：2019-02-28 09:54

【摘要】：间充质干细胞(Mesenchymal stem cells, MSCs)是干细胞家族中具有多向分化潜能以及自我更新能力的成体干细胞。自2002年Woodbury等人首次报道将骨髓MSCs体外诱导分化成神经元以来,目前体外诱导MSCs分化成神经元细胞有4种方法：抗氧化剂诱导、细胞因子诱导、中药诱导以及基因编辑诱导。如何使MSCs分化而来的类神经元细胞具有神经细胞的特性,是我们探究的方向。Ω-3及Ω-6族多不饱和脂肪酸(polyunsaturated fatty acid, PUFA)及其代谢产物与细胞结构及功能有关,它们可以通过多种作用机制,促进干细胞的增殖与分化,继而影响干细胞的命运。我们的实验在人脐带间充质干细胞(Human umbilical cord mesenchymal stem cells, hUC-MSCs)神经分化培养体系中增补PUFA,包括二十碳五烯酸(Eicosapentaenoic acid, EPA)及二十二碳五烯酸(Docosapentenoic acid, DPA), MTS assay以及Transwell迁移实验结果表明,EPA和DPA可以促进hUC-MSCs的增殖、迁移；而qRT-PCR以及免疫细胞化学检测表明,增补EPA和DPA后hUC-MSCs分化的类神经元细胞中多种神经元特异性标记的表达高于未增补PUFA的对照组,包括胶质纤维酸性蛋白(Glial fibrillary acidic protein,GFAP)、神经丝蛋白(Neurofilament, NF)、酪氨酸羟化酶(Tyrosine-hydroxylase,TH)、神经元核抗原(Neuronal nuclear antigen, NeuN)、神经元特异性稀醇化酶(Neuron-specific enolase, NES)以及微管结合蛋白2(Microtubule associated protein 2, MAP2)等。这一结果显示EPA和DPA参与hUC-MSCs增殖、迁移及神经分化。本实验的研究成果有望用于细胞替代治疗中,多不饱和脂肪酸增补培养hUC-MSCs后,分化成可行使功能的神经细胞,替代引起病变的已坏死的神经细胞,以达到减缓或治愈神经退行性疾病的目的。
[Abstract]:Mesenchymal stem cells (Mesenchymal stem cells, MSCs) are adult stem cells with multi-directional differentiation potential and self-renewal ability in stem cell family. Since Woodbury and others first reported the differentiation of bone marrow MSCs into neurons in vitro, there are four methods to induce MSCs to differentiate into neurons in vitro: antioxidant induction, cytokine induction, traditional Chinese medicine induction and gene editing induction. How to make the neuron-like cells differentiated from MSCs have the characteristics of neural cells is the research direction. 惟-3 and 惟-6 polyunsaturated fatty acids (polyunsaturated fatty acid, PUFA) and their metabolites are related to the structure and function of cells. They can promote the proliferation and differentiation of stem cells through a variety of mechanisms, and then affect the fate of stem cells. In our experiment, we added PUFA, to the neural differentiation culture system of human umbilical cord mesenchymal stem cells (Human umbilical cord mesenchymal stem cells, hUC-MSCs), including eicosapentaenoic acid (Eicosapentaenoic acid, EPA) and 22 carbapentaenoic acid (Docosapentenoic acid, DPA),). The results of MTS assay and Transwell migration showed that EPA and DPA could promote the proliferation and migration of hUC-MSCs. The results of qRT-PCR and immunocytochemistry showed that the expression of many neuron specific markers in hUC-MSCs differentiated neuron cells supplemented with EPA and DPA was higher than that in the control group without PUFA, including glial fibrillary acidic protein (Glial fibrillary acidic protein,. GFAP), neurofilament protein (Neurofilament, NF), tyrosine hydroxylase (Tyrosine-hydroxylase,TH), neuronal nuclear antigen (Neuronal nuclear antigen, NeuN), neuron specific dilute alcoholase (Neuron-specific enolase, NES) and tubulin binding protein 2 (Microtubule associated protein 2, MAP2, etc. These results suggest that EPA and DPA are involved in the proliferation, migration and neural differentiation of hUC-MSCs. The results of this study are expected to be used in cell replacement therapy. After adding polyunsaturated fatty acids to cultured hUC-MSCs, polyunsaturated fatty acids can differentiate into functional nerve cells and replace necrotic neurons that cause pathological changes. In order to alleviate or cure neurodegenerative diseases.
【学位授予单位】：内蒙古大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R329.2

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