BMSCs向胰岛β-细胞分化机理及其分化细胞治疗犬糖尿病可行性研究

Diabetes has become the third major disease harming the health of humans and animals. Islet transplantation is an ideal treatment for the disease, but it is hampered by donor deficiencies and immune rejection. Bone marrow mesenchymal stem cells (BMSCs) have multiple differentiation potential, low immunogenicity and immunosuppressive capacity after transplantation, are ideal seed cells for the treatment of diabetes. However, the differentiation mechanism of BMSCs to islet β-cells is unclear for now, which results in very low differentiation efficiency of BMSCs induced in vitro and immature differentiated-cells. Furthermore, present studies about transplantation of BMSCs differentiated-cells for treatment of diabetes were only seen in rodent animals, which is not enough to offer references for medical clinical application of large animals and humans. In this study, canine BMSCs are intended to use as seed cells, high-throughput sequencing would be used to detect the expression changes of the new differentially expressed genes and the key genes in the chain cascade during the differentiation of BMSCs to islet β-cells, and to verify the relationship between them, and finally to reveal the differentiation mechanism of BMSCs to islet β-cells in vitro. Then the induction methods and procedures in vitro will be redesigned and optimized based on the mechanism to improve the differentiation efficiency and maturity of differentiated cells, and the feasibility inducing BMSCs into islet β-cells in large quantities in vitro and transplanting differentiated cells to treat canine diabetes would be studied, which would provide a new method for the clinical treatment of animal diabetes and large-scale animal experimental data and models for the treatment of human diabetes.

糖尿病已成为危害人和动物身体健康的第三大疾病，胰岛移植是治疗该病的理想方法，但因供体缺乏和免疫排斥而受阻；骨髓间充质干细胞（BMSCs）免疫原性低、对受体淋巴细胞有抑制能力，是治疗糖尿病最理想的种子细胞，但因BMSCs向胰岛β-细胞分化机理不清，致使其诱导分化效率很低；再者，采用BMSCs分化细胞治疗糖尿病研究仅见于啮齿动物，不足以为大动物和人医临床应用提供参考。本研究拟以犬BMSCs作种子细胞，采用高通量测序技术检测BMSCs向胰岛β-细胞分化过程中新差异表达基因和链式级联调控系统中关键基因的表达变化，分析和验证他们时序表达和相互调控关系，揭示体外诱导下BMSCs向胰岛β-细胞分化机理；依据机理重新制定和优化体外诱导方法和程序，提高其分化效率；进而研究大批量诱导BMSCs向胰岛β-细胞分化和治疗犬糖尿病的可行性，为兽医糖尿病治疗提供新方法，为人医提供大动物试验资料和模型。

糖尿病已成为危害人和动物身体健康的第三大疾病，胰岛移植是治疗该病的理想方法，但因供体缺乏和免疫排斥而受阻；间充质干细胞来源广泛，免疫原性低，且有免疫抑制能力，是治疗糖尿病最理想的种子细胞，但因间充质干细胞向胰岛 β 细胞分化机理不清，致使其诱导分化效率很低，不能提供临床应用。本研究以犬 间充质干细胞作种子细胞，采用高通量测序技术检测了其向胰岛 β 细胞分化过程中新差异表达基因和链式级联调控系统中关键基因的表达变化，分析和验证了他们时序表达和相互调控关系，揭示了体外诱导下间充质干细胞 向胰岛 β 细胞分化机理；并且依据机理重新制定和优化了体外诱导方法和程序，提高了分化效率；此外，成功制备了犬Ⅰ型糖尿病模型，证明了新生胰岛 β 细胞和间充质干细胞共移植治疗Ⅰ型糖尿病模型犬是可行的。本项目为间充质干细胞治疗犬糖尿病奠定了基础，为人医糖尿病细胞替代治疗提供了参考。