Kidins220在胚胎干细胞中的功能及人胚胎干细胞来源间质祖细胞治疗小鼠急性肾损伤研究

发布时间：2018-02-17 06:07

本文关键词： 胚胎干细胞 Kidins220/ARMS 间质祖细胞急性肾损伤细胞治疗　出处：《浙江大学》2012年博士论文　论文类型：学位论文

【摘要】：胚胎干细胞来源于哺乳动物囊胚内细胞团。它在体外培养条件下可以保持自我更新能力和多向分化潜能,是体外发育生物学研究的理想模型,并为再生治疗提供了细胞来源。对促进胚胎干细胞增殖的分子机理的研究是胚胎干细胞生物学研究的核心命题。胚胎干细胞的自我更新和分化受到胞外信号分子和胞内信号通路的协同调控。作为两者的介导者,细胞膜表面的跨膜蛋白对胚胎干细胞的自我更新起着关键作用。因此,对新的胚胎干细胞表面功能性分子的筛选有助于深入理解干细胞增殖相关的调控机理。 Kidins220/ARMS是神经营养因子信号通路的关键调节蛋白,对神经细胞的存活和增殖起到重要作用。但是Kidins220/ARMS在胚胎干细胞上的表达及功能尚未进行研究。本实验室通过对小鼠胚胎干细胞膜表面蛋白质组学数据分析,结合实时定量PCR和ICC鉴定,在胚胎干细胞上首次筛选得到一种高丰度表达的膜表面蛋白Kidins220/ARMS.利用RNA干扰技术,胚胎干细胞中Kidins220/ARMS的表达下调。进一步研究证明Kidins220/ARMS的下调并未影响到小鼠胚胎干细胞的自我更新,但是显著抑制了它的增殖。这说明Kidins220/ARMS对于维持胚胎干细胞正常的增殖非常重要。对Kidins220/ARMS影响胚胎干细胞增殖相关的分子机理作进一步研究有助于加深对干细胞增殖机理的理解,建立新的ESCs培养体系,理解Kidins220/ARMS在小鼠胚胎发育中的生物学意义。急性肾损伤(AKI)是一种严重的临床疾病,具有较高的致残率和致死率。虽然很多药物治疗方法,包括钙通道阻止剂,多巴胺、内皮素的佶抗剂和利尿排泄多肽等已经进入临床实验,但成功率较低,并伴有多种副作用。因此,干细胞移植为急性肾损伤的治疗提供了新的希望,已经有越来越多的研究者将各种来源的干细胞应用于疾病治疗。本实验室通过人胚胎干细胞的定向分化,获得了一种新的干细胞,人胚胎干细胞来源的间质祖细胞(hESC-MPs).它具有自我更新能力,可以向成脂、成骨和成软骨细胞分化。我们首次将hESC-MPs应用于急性肾损伤的治疗。将hESC-MPs移植到顺铂(Cisplatin)诱导的急性肾损伤小鼠模型中,显著改善了受损伤小鼠的肾功能,促进了肾脏组织结构的修复。对人特异的17号染色体α微卫星片段的PCR检测揭示移植的hESC-MPs归巢到了肾脏的损伤区域,促进了肾细胞的增殖和存活。而且,hESC-MPs的移植活化了抗炎症因子IL-10和促存活因子bFGF、FIGF、IGF-1的表达,降低了趋化因子MCP-1和促炎症因子IL-1β、TNF-α、IFN-γ、IL-6和TGF-β的基因表达量。这提示hESC-MPs的免疫调节属性在急性肾损伤的修复中起到了重要作用。这些结果揭示hESC-MPs是一种非常有希望的细胞来源,可用于急性肾损伤和其他复杂的多因素失调性疾病的治疗。
[Abstract]:Embryonic stem cells come from the inner cell mass of mammalian blastocysts. It can maintain self-renewal and multi-differentiation potential in vitro, and is an ideal model for the study of developmental biology in vitro. The molecular mechanism of promoting the proliferation of embryonic stem cells is the core proposition in the biological research of embryonic stem cells. The self-renewal and differentiation of embryonic stem cells are subject to extracellular signaling molecules. And intracellular signaling pathways. As mediators of both, The transmembrane proteins on the membrane surface play a key role in the self-renewal of embryonic stem cells. Therefore, the screening of new functional molecules on the surface of embryonic stem cells is helpful to further understand the regulatory mechanisms related to stem cell proliferation. Kidins220/ARMS is a key regulator of neurotrophic factor signaling pathway. However, the expression and function of Kidins220/ARMS on embryonic stem cells have not been studied. In combination with real-time quantitative PCR and ICC identification, a highly abundant membrane surface protein Kidins220 / ARMSwas obtained from embryonic stem cells for the first time. RNA interference technique was used. Kidins220/ARMS expression was down-regulated in embryonic stem cells. Further studies showed that the down-regulation of Kidins220/ARMS did not affect the self-renewal of mouse embryonic stem cells. But it significantly inhibited its proliferation. This suggests that Kidins220/ARMS is very important for maintaining the normal proliferation of embryonic stem cells. Further study of the molecular mechanism related to the effects of Kidins220/ARMS on the proliferation of embryonic stem cells will contribute to a better understanding of the mechanism of stem cell proliferation. To establish a new ESCs culture system to understand the biological significance of Kidins220/ARMS in mouse embryo development. Acute renal injury (AKI) is a serious clinical disease with a high rate of disability and mortality. Although many drug treatments include calcium channel blockers, dopamine, Et has been used in clinical trials with low success rate and many side effects. Therefore, stem cell transplantation offers new hope for the treatment of acute renal injury. More and more researchers have applied stem cells from various sources to the treatment of diseases. Through the directional differentiation of human embryonic stem cells, we have obtained a new kind of stem cells, human embryonic stem cell derived mesenchymal progenitor cells (hESC-MPsN). Osteogenesis and chondroblast differentiation. We used hESC-MPs for the first time in the treatment of acute renal injury. Transplantation of hESC-MPs into the model of acute renal injury induced by cisplatin significantly improved the renal function of injured mice. The PCR analysis of the specific human chromosome 17 伪 microsatellite fragment revealed that the transplanted hESC-MPs homed to the damaged area of the kidney. Moreover, the transplantation of hESC-MPs activates the expression of anti-inflammatory factor IL-10 and survivor-promoting factor bFGFFIGF-1, and promotes the proliferation and survival of renal cells, and the transplantation of hESC-MPs activates the expression of anti-inflammatory factor IL-10 and survivor-promoting factor bFGFFIGF-1. These results suggest that the immunomodulatory properties of hESC-MPs play an important role in the repair of acute renal injury. These results suggest that hESC-MPs is a very promising cell source. It can be used in the treatment of acute renal injury and other complex multifactorial disorders.
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R329

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