阿尔茨海默病相关基因白细胞介素18（IL18）启动子区多态性对其转录活性的影响
发布时间：2018-11-28 19:20
【摘要】：【研究背景】阿尔茨海默病(Alzheimer's disease, AD)是一种主要在老年期发生的以进行性痴呆为主要特征的神经元退行性疾病。AD的病因尚未明确，众多研究表明其与脑内慢性炎症反应相关。IL18作为一种炎症细胞因子，近年越来越多被证实与AD发病相关。IL18基因调控区存在－607C/A（RS1946518）、－137G/C（RS187238）等SNPs，已有文献报道多态性与老年性痴呆发病间的关系，但未见这两处基因多态性对IL18基因转录活性影响的报道。 【研究目的】本课题通过分子生物学技术，建立IL18－607C/A、IL18－137G/C野生型和突变等位基因的PGL4.10重组试验质粒，在细胞水平研究－607C/A突变、－137G/C突变对IL18基因转录调控的影响。 【方法】选取双荧光报告基因系统（pGL4.10（luc2) Basic及pRL-TK)作为报告基因载体，用基因重组和定点突变技术构建含不同SNP型的IL18启动子报告基因质粒，转染人胚胎肾细胞系293细胞（HEK293)，计算不同SNP各启动子的相对荧光素酶活性（relative luciferase activity，，RLA) 【结果】所构建的各SNP型质粒均具有明确的启动子活性；突变质粒-607A/-137C、-607C/-137G及-607C/-137C与野生质粒-607A/-137G相比，转录活性均有显著性差异，（P0.01）；而单位点突变质粒-607A/-137C、-607C/-137G与两位点突变质粒-607C/-137C转录活性相比均无差异性(P0.05)。 【结论】-607、-137位点碱基的突变均可影响IL18基因的转录，-607A/-137C、-607C/-137G及-607C/-137C均上调了IL18基因的转录活性，致IL18蛋白表达水平增加，促进神经炎症的病理过程，可能导致AD的发生。
[Abstract]:[background] Alzheimer's disease (Alzheimer's disease, AD) is a neuronal degenerative disease characterized mainly by progressive dementia in the elderly. The etiology of AD is not clear. IL18, as an inflammatory cytokine, has been more and more associated with the pathogenesis of AD in recent years. SNPs, exists in the regulatory region of IL18 gene such as 607C/A (RS1946518),-137G/C (RS187238) and so on. The relationship between polymorphism and Alzheimer's disease has been reported, but there is no report on the effect of these two gene polymorphisms on the transcriptional activity of IL18 gene. [objective] by using molecular biology technique, the PGL4.10 recombinant plasmid of IL18-607C/A,IL18-137G/C wild-type and mutant alleles was established, and the 607C/A mutation was studied at the cell level. The effect of 137G/C mutation on the transcriptional regulation of IL18 gene. [methods] double fluorescent reporter gene systems (pGL4.10 (luc2) Basic and pRL-TK) were selected as reporter gene vectors. The IL18 promoter reporter gene plasmids containing different SNP types were constructed by gene recombination and site-directed mutagenesis. Transfected into human embryonic kidney cell line 293 (HEK293), the relative luciferase activity (relative luciferase activity,RLA) of different SNP promoters was calculated. [results] all the SNP type plasmids constructed showed definite promoter activity. The transcriptional activities of mutant plasmids -607A / -137C / -607C / -137G and-607C/-137C were significantly different from those of wild plasmids-607A/-137G (P0.01). However, the transcriptional activity of the unit point mutant plasmide-607A / -137C / -607C / -137G was not different from that of the two locus mutant plasmids-607C/-137C (P0.05). [conclusion] the mutation at -607C ~ 137 can affect the transcription of IL18 gene. -607A / -137C / -137C / -137G and-607C/-137C can up-regulate the transcriptional activity of IL18 gene and increase the expression of IL18 protein. Promoting the pathological process of neuroinflammation may lead to the occurrence of AD.
【学位授予单位】：广州医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R749.16
本文编号：2364028
[Abstract]:[background] Alzheimer's disease (Alzheimer's disease, AD) is a neuronal degenerative disease characterized mainly by progressive dementia in the elderly. The etiology of AD is not clear. IL18, as an inflammatory cytokine, has been more and more associated with the pathogenesis of AD in recent years. SNPs, exists in the regulatory region of IL18 gene such as 607C/A (RS1946518),-137G/C (RS187238) and so on. The relationship between polymorphism and Alzheimer's disease has been reported, but there is no report on the effect of these two gene polymorphisms on the transcriptional activity of IL18 gene. [objective] by using molecular biology technique, the PGL4.10 recombinant plasmid of IL18-607C/A,IL18-137G/C wild-type and mutant alleles was established, and the 607C/A mutation was studied at the cell level. The effect of 137G/C mutation on the transcriptional regulation of IL18 gene. [methods] double fluorescent reporter gene systems (pGL4.10 (luc2) Basic and pRL-TK) were selected as reporter gene vectors. The IL18 promoter reporter gene plasmids containing different SNP types were constructed by gene recombination and site-directed mutagenesis. Transfected into human embryonic kidney cell line 293 (HEK293), the relative luciferase activity (relative luciferase activity,RLA) of different SNP promoters was calculated. [results] all the SNP type plasmids constructed showed definite promoter activity. The transcriptional activities of mutant plasmids -607A / -137C / -607C / -137G and-607C/-137C were significantly different from those of wild plasmids-607A/-137G (P0.01). However, the transcriptional activity of the unit point mutant plasmide-607A / -137C / -607C / -137G was not different from that of the two locus mutant plasmids-607C/-137C (P0.05). [conclusion] the mutation at -607C ~ 137 can affect the transcription of IL18 gene. -607A / -137C / -137C / -137G and-607C/-137C can up-regulate the transcriptional activity of IL18 gene and increase the expression of IL18 protein. Promoting the pathological process of neuroinflammation may lead to the occurrence of AD.
【学位授予单位】：广州医学院
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R749.16
【参考文献】
相关期刊论文 前5条
1 罗文浩,罗韵文,黄本友;丘脑梗死对记忆和智能的影响[J];临床内科杂志;2002年02期
2 彭丹涛;许贤豪;张振馨;魏镜;王鲁宁;谢恒革;王荫华;贾建平;于新;;阿尔茨海默病各种临床记忆检测方法的评价[J];中华老年心脑血管病杂志;2007年05期
3 周爱红;贾建平;闵宝权;杨培洁;;轻度血管性认知障碍神经心理评估量表的构建[J];中华老年心脑血管病杂志;2008年04期
4 李永秋,张昱,马建国;丘脑梗塞的临床与CT对照研究[J];中风与神经疾病杂志;1998年03期
5 周礼亮;高洁;;丘脑卒中患者认知功能改变的临床观察[J];中国神经免疫学和神经病学杂志;2009年01期
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