基因甲基化与原发性痛风的相关性研究

发布时间：2018-10-12 15:21

【摘要】：目的:探讨基因(ABCG2、SLC2A9和P2RX7)CpG岛处甲基化与原发性痛风的相关性;并应用iTRAQ标记技术结合液相色谱串联质谱技术,筛选出原发性痛风血清差异蛋白,进一步分析差异蛋白(TBB4A)的编码基因(TUBB4A)的甲基化情况,为原发性痛风的早期诊断和早期治疗提供了坚实的理论依据。方法:1、提取原发性痛风组及健康对照组的全血DNA,应用焦磷酸测序方法进行基因甲基化检测。用t检验或非参数检验进行甲基化水平的分析。2、应用iTRAQ标记技术结合液相色谱串联质谱技术筛选出差异蛋白,并且通过生物学过程、细胞定位、分子功能、KEGG通路及蛋白互作上进行分析生物信息学分析找出有意义的差异蛋白。3、用Elisa技术进行差异蛋白的血清学检测,BCA法进行总蛋白的血清学检测,用t检验或非参数检验进行差异蛋白含量的分析。结果:1、ABCG2 CpG(pos1、2、4、5、6),SLC2A9 CpG(pos1、3、4、5、7)及P2RX7CpG pos2甲基化水平差异有统计学意义(均P0.05)。2、共鉴定出95个差异蛋白(其中50个上调蛋白和45个下调蛋白)和20条显著的KEGG通路。其中,甘油醛-3-磷酸脱氢酶(GAPDH)、α-烯醇酶(ENOA)、磷酸甘油酸激酶(PGK1)、葡萄糖-6-磷酸异构酶(G6PI)和膜突蛋白(MOES)等差异蛋白在原发性痛风发病中可能起到重要的作用。3、蛋白TBB4A在原发性痛风的血清中明显高于健康对照组(P=0.023)。4、TUBB4A CpG pos4甲基化水平差异有统计学意义(P=0.036)。结论:1、基因ABCG2、SLC2A9和P2RX7的甲基化水平与原发性痛风的发病相关,但基因ABCG2的作用机制并未明确。2、蛋白TBB4A在血清中的表达与原发性痛风的发病相关。3、基因TUBB4A的甲基化水平与原发性痛风的发病相关,但其作用机制仍不明。
[Abstract]:Objective: to investigate the correlation between methylation at CpG island of gene (ABCG2,SLC2A9 and P2RX7) and primary gout, and to screen serum differential proteins of primary gout by iTRAQ labeling and liquid chromatography-tandem mass spectrometry (LC-MS). Further analysis of methylation of differential protein (TBB4A) encoding gene (TUBB4A) provides a solid theoretical basis for early diagnosis and early treatment of primary gout. Methods: 1. Gene methylation was detected by pyrosequencing method in whole blood DNA, extracted from primary gout group and healthy control group. T test or nonparametric test was used to analyze methylation level. 2. Differential proteins were screened by iTRAQ labeling and liquid chromatography-tandem mass spectrometry (LC-MS). Molecular function, KEGG pathway and protein interaction were analyzed by bioinformatics to find the significant differential protein. 3. The differential protein was serologically detected by Elisa, and the total protein was detected by BCA. T-test or non-parametric test was used to analyze the differential protein content. Results: 1 the levels of ABCG2 CpG (pos1,2,4,5,6), SLC2A9 CpG (pos1,3,4,5,7) and P2RX7CpG pos2 methylation were significantly different (P0.05). A total of 95 differentially expressed proteins (50 up-regulated proteins and 45 down-regulated proteins) and 20 KEGG pathways were identified. Of which, Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), 伪 -enolase (ENOA), phosphoglycerate kinase (PGK1), glucose-6-phosphate isomerase (G6PI) and membranous protein (MOES) may play an important role in the pathogenesis of primary gout. The serum methylation level of TUBB4A in wind group was significantly higher than that in healthy control group (P0. 023), and the level of CpG pos4 methylation in TUBB4A was significantly higher than that in healthy control group (P0. 036). Conclusion: 1 the methylation level of gene ABCG2,SLC2A9 and P2RX7 is associated with the pathogenesis of primary gout. However, the mechanism of gene ABCG2 is not clear, the expression of protein TBB4A in serum is related to the pathogenesis of primary gout, the methylation level of gene TUBB4A is related to the pathogenesis of primary gout, but the mechanism is still unclear.
【学位授予单位】：宁波大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R589.7

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