方格星虫多能性因子预测及其体表损伤修复的表达研究
发布时间：2018-10-24 09:47
【摘要】：干细胞多能性因子是一类能将体细胞诱导成多能干细胞的转录因子,诱导性多能干细胞与胚胎干细胞极其相似。多能性因子的概念一经提出,其与损伤修复和再生的联系就倍受关注。作为进入重编程的关键因素,其拥有的特质与组织的再生关系密切。在各门类物种中,探索多能性因子与损伤修复和再生的联系,有利于进一步确认多能性因子在再生过程中所处的重要地位,为再生医学的研究提供理论依据。方格星虫进化地位较低,然而其机体损伤修复的过程与高等生物非常相似。利用方格星虫探索多能性因子与损伤修复和再生的联系具有理论意义。本课题将立足于多能性因子,探索其与方格星虫体表损伤修复之间的联系。本课题基于方格星虫转录组的测序结果,进行初步分析,发现24种候选干细胞多能性因子在方格星虫中表达了17种,4种在诱导过程中发挥关键性作用的干细胞多能性因子,在方格星虫中发现了3种,分别是Sox2、c-Myc和Klf4。另外Fthl处于所有组织内的表达水平远远强于别的多能性因子。同时发现在方格星虫中存在4条调控多能性的途径。方格星虫多能性因子同源基因与其再生相关同源基因在部分组织中的表达趋势也存在相似性。根据预测结果从中选取出4种因子,Sox2、Klf4、c-Myc、Fthl以及WNT信号通路中的Wnt-7a、beta-Catenin,进行损伤修复的功能验证。采用手术刀对方格星虫肛门对侧的体壁肌肉组织进行纵向划伤,伤口长度约1 cm,深度约1-2 mm,损伤后分别在0 h、10 min、30 min、12 h、24 h、48 h、72 h采集损伤组织进行功能验证,采用qPCR与免疫组化技术评估6种基因在损伤修复过程中表达量的变化以及组织细胞的增殖情况。实验数据表明,在方格星虫体表损伤修复过程中,这6种基因均能够响应损伤,并且Sox2、Klf4、Fthl、Wnt-7a和beta-Catenin在损伤修复过程中,能够明显的呈现出表达量的上调。从损伤后24 h开始,伤口附近的组织开始出现增殖的现象。根据实验结果可以说明,在方格星虫中,多能性因子与损伤修复确实存在着正相关的联系。结合蚯蚓、蝾螈已有的研究结果推测,多能性因子与再生的联系会随着再生能力的增强而变得更加紧密。
[Abstract]:Stem cell pluripotent factor is a kind of transcription factor which can induce somatic cells into pluripotent stem cells. Induced pluripotent stem cells are very similar to embryonic stem cells. Once the concept of pluripotent factor is put forward, its relation with damage repair and regeneration has attracted much attention. As a key factor in reprogramming, its characteristics are closely related to organizational regeneration. To explore the relationship between pluripotent factors and injury repair and regeneration in various species is helpful to confirm the important position of pluripotent factors in regeneration process and to provide theoretical basis for the research of regenerative medicine. The phylogenetic status is low, but the process of damage repair is very similar to that of higher organisms. It is of theoretical significance to explore the relationship between pluripotent factors and damage repair and regeneration by using the starfish. Based on the pluripotent factor, this paper will explore the relationship between it and the repair of surface damage. Based on the sequencing results of the transcriptome, we found that 24 candidate stem cell pluripotent factors expressed 17 species and 4 stem cell pluripotent factors played a key role in the induction process. Three species, Sox2,c-Myc and Klf4., have been found in this field. In addition, the expression level of Fthl in all tissues is much stronger than other pluripotent factors. At the same time, there are 4 ways to regulate the pluripotency in the starfish. There was also similarity in the expression trend between the homologous genes of the multipotent factors and their regenerative homologous genes in some tissues. According to the predicted results, four factors, Sox2,Klf4,c-Myc,Fthl and Wnt-7a,beta-Catenin, in WNT signaling pathway, were selected to verify the function of damage repair. The muscle tissue of the opposite side of the anus of Scopodium oblongata was scratched longitudinally by scalpel. The wound length was about 1 cm, and the depth of injury was about 1-2 mm,. The injured tissues were collected at 0 h 10 min,30 min,12 h ~ 24 h and 48 h ~ 72 h for functional verification. QPCR and immunohistochemical techniques were used to evaluate the expression of six genes and the proliferation of histocytes in the process of injury and repair. The experimental data showed that the six genes were able to respond to the damage in the process of surface damage repair, and the expression of Sox2,Klf4,Fthl,Wnt-7a and beta-Catenin were up-regulated during the process of damage repair. From 24 hours after injury, the tissue around the wound began to proliferate. According to the experimental results, there is a positive correlation between pluripotent factors and damage repair. Combined with earthworm and salamander, it is speculated that the relationship between pluripotent factor and regeneration will become closer with the increase of regeneration ability.
【学位授予单位】：华侨大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R318;Q78
本文编号：2291035
[Abstract]:Stem cell pluripotent factor is a kind of transcription factor which can induce somatic cells into pluripotent stem cells. Induced pluripotent stem cells are very similar to embryonic stem cells. Once the concept of pluripotent factor is put forward, its relation with damage repair and regeneration has attracted much attention. As a key factor in reprogramming, its characteristics are closely related to organizational regeneration. To explore the relationship between pluripotent factors and injury repair and regeneration in various species is helpful to confirm the important position of pluripotent factors in regeneration process and to provide theoretical basis for the research of regenerative medicine. The phylogenetic status is low, but the process of damage repair is very similar to that of higher organisms. It is of theoretical significance to explore the relationship between pluripotent factors and damage repair and regeneration by using the starfish. Based on the pluripotent factor, this paper will explore the relationship between it and the repair of surface damage. Based on the sequencing results of the transcriptome, we found that 24 candidate stem cell pluripotent factors expressed 17 species and 4 stem cell pluripotent factors played a key role in the induction process. Three species, Sox2,c-Myc and Klf4., have been found in this field. In addition, the expression level of Fthl in all tissues is much stronger than other pluripotent factors. At the same time, there are 4 ways to regulate the pluripotency in the starfish. There was also similarity in the expression trend between the homologous genes of the multipotent factors and their regenerative homologous genes in some tissues. According to the predicted results, four factors, Sox2,Klf4,c-Myc,Fthl and Wnt-7a,beta-Catenin, in WNT signaling pathway, were selected to verify the function of damage repair. The muscle tissue of the opposite side of the anus of Scopodium oblongata was scratched longitudinally by scalpel. The wound length was about 1 cm, and the depth of injury was about 1-2 mm,. The injured tissues were collected at 0 h 10 min,30 min,12 h ~ 24 h and 48 h ~ 72 h for functional verification. QPCR and immunohistochemical techniques were used to evaluate the expression of six genes and the proliferation of histocytes in the process of injury and repair. The experimental data showed that the six genes were able to respond to the damage in the process of surface damage repair, and the expression of Sox2,Klf4,Fthl,Wnt-7a and beta-Catenin were up-regulated during the process of damage repair. From 24 hours after injury, the tissue around the wound began to proliferate. According to the experimental results, there is a positive correlation between pluripotent factors and damage repair. Combined with earthworm and salamander, it is speculated that the relationship between pluripotent factor and regeneration will become closer with the increase of regeneration ability.
【学位授予单位】：华侨大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R318;Q78
【参考文献】
相关期刊论文 前3条
1 沈先荣,蒋定文,贾福星,储智勇,周菊珍,陈美华;海洋星虫提取物的抗疲劳作用研究[J];中华航海医学与高气压医学杂志;2003年02期
2 李凤鲁,孔庆兰,史贵田,王玮,周红,金善福;中国沿海方格星虫属(星虫动物门)的研究[J];青岛海洋大学学报;1990年01期
3 李复雪;高世和;周时强;;福建沿海红树林区的动物资源及其开发利用[J];福建水产;1989年04期
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