PDGFRa在人神经管畸形胚胎中的表达

发布时间：2018-05-02 22:32

本文选题：神经管畸形(NTDs) + 血小板源性生长因子受体　；参考：《山西医科大学》2005年硕士论文

【摘要】：目的应用全基因组表达芯片研究山西吕梁地区神经管畸形(neural tube defects,NTDs)胚胎脑组织中状念特异性表达基因;将差异表达基因从四氢叶酸代谢水平、基因表达调控蛋白、信号转导途径、凋亡等途径分类;进一步研究于神经系统发育相关的血小板源性生长因子受体α(platelet-derived growth factor receptor-alpha PDGFRα)及其调节因子PAX1、PAX3-FKHR、EGFR在神经管畸形疾病中的作用机制。方法利用全基因组表达芯片高通量筛选出NTDs各发病阶段的异常表达基因;并从生化与分子生物学的角度加以归类,揭示出NTDs的异常基因表达谱;从中选择出PDGFRα基因应用半定量RT-PCR技术扩增PDGFRα及其调节因子PAX1、PAX3-FKHR、EGFR并验证其改变,同时应用免疫组化分析PDGFRα在蛋白质水平的表达。结果芯片结果显示出异常表达的基因22209个,其中2倍表达增高的基因74个,包括氧化磷酸化途径中相关基因3个,锌指结构基因7个,与抗氧化谷光甘肽通路基因2个,与花生四烯酸合成代谢相关的基因2个,MMPs相关的基因4个,G蛋白膜受体家族相关基因6个等。2倍表达降低的387个,其中与核酸代谢通路相关基因8个,炎症反应相关基因7个,参与凋亡途径的基因6个,整合素家族信号系统相关基因2个,三羧酸循环通路相关基因4个,G蛋白膜受体家族相关基因9个等,PDGFRα呈2~(0.9)倍增高;半定量RT-PCR结果显示NTDs胚胎中PDGFRα表达高于对照组(p0.01)结果显示NTDs胚胎中其调节因子PAX1、PAX3-FKHR、EGFR表达高于对照组(p0.01),免疫组化结果显示NTDs组PDGFRα蛋白表达高于对照组(p0.05)。结论基因芯片研究结果显示NTDs脑组织与对照脑组织的基因表达谱相比存在很大差别,证明NTDs的发生和发展是一个复杂的多阶段的过程,是多种相关基因表达失常所致,涉及核酸代谢、信号转导、基因表达调控、凋亡等基因的异常改变;PDGFRα调节细胞的生长、存活,并且对细胞的形态和细胞运动如细胞的分散、趋化、聚集、凋亡等有作用。PDGFRα的过度表达干扰了神经上皮细胞正常的分化、迁移和凋亡,导致膜信号转导系统紊乱,凋亡信号减弱,神经上皮某些细胞持续生长不分化(神经胶质、软骨前细胞等),继而影响正
[Abstract]:Objective to study the expression of specific genes in embryonic brain of neural tube defectsus NTDsin Lv Liang area of Shanxi province by using the whole genome expression chip, and to express the differentially expressed genes from the level of tetrahydrofolic acid metabolism and the expression of regulatory proteins. The signal transduction pathway and apoptosis pathway were classified, and the mechanism of PAX1-PAX3-FKHR-EGFR in neural tube malformation was further studied, which was related to the development of the nervous system, platelet-derived growth factor receptor-alpha PDGFR 伪) and its regulatory factor PAX1-PAX3-FKHR-EGFR. Methods the abnormal expression genes of NTDs were screened by high throughput genomic expression microarray, and the abnormal gene expression profiles of NTDs were revealed from biochemical and molecular biology. The PDGFR 伪 gene was selected to amplify PDGFR 伪 and its regulatory factor PAX1-PAX3-FKHR-EGFR by semi-quantitative RT-PCR technique. The expression of PDGFR 伪 at protein level was analyzed by immunohistochemistry. Results the microarray results showed that there were 22209 abnormal expression genes, among which 74 genes were double expressed, including 3 genes related to oxidative phosphorylation pathway, 7 genes related to zinc finger structure, and 2 genes related to antioxidation glutamate pathway. There were 4 genes related to arachidonic acid biosynthesis and 4 genes related to MMPs, and 387 genes whose expression was reduced by 2.2-fold, including 8 genes related to nucleic acid metabolism pathway and 7 genes related to inflammation. There were 6 genes involved in apoptotic pathway, 2 integrin family signal system related genes, 4 tricarboxylic acid cycle pathway related genes, and 9 G protein membrane receptor family related genes. The results of semi-quantitative RT-PCR showed that the expression of PDGFR 伪 in NTDs embryos was higher than that in control group (p0.01). The expression of PAX1-PAX3-FKHR-EGFR in NTDs embryos was higher than that in control group, and the expression of PDGFR 伪 protein in NTDs group was higher than that in control group. Conclusion the gene expression profile of NTDs brain tissue is different from that of control brain tissue, which indicates that the occurrence and development of NTDs is a complicated and multistage process, which is caused by the abnormal expression of many related genes. Abnormal changes in genes involved in nucleic acid metabolism, signal transduction, gene expression regulation, apoptosis, etc. PDGFR 伪 regulates cell growth, survival, and cell morphology and cell movement such as cell dispersion, chemotaxis and aggregation. The overexpression of PDGFR 伪 interferes with the normal differentiation, migration and apoptosis of neural epithelial cells, which leads to the disturbance of membrane signal transduction system, the weakening of apoptotic signal, and the persistent growth and undifferentiation of some cells of neural epithelium (glia, glia, glia, glia, etc.) Prechondral cells, etc., which in turn affect the positive
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R361

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