缺氧诱导脂滴相关蛋白在糖尿病合并腹主动脉瘤中的表达

Expression of hypoxia-inducible lipid droplet-associated protein in diabetic patients combined with abdominal aortic aneurysm

摘要:

目的 通过生物信息学方法,寻找糖尿病合并腹主动脉瘤(AAA)患者中差异表达的基因,明确其功能,探讨发病机制。方法 从Gene Expression Omnibus(GEO)数据库中下载人糖尿病GSE21321和人腹主动脉瘤GSE7084数据集,以P＜0.05及表达变化＞2.0倍标准筛选差异基因。联合运用Gencript数据库和predictprotein平台分析蛋白质亚细胞定位,并通过荧光共定位实验进行验证；STRING数据库分析缺氧诱导脂滴相关蛋白(HILPDA)的相互作用。采用 SPSS 13.0 软件进行数据分析,计量资料差异比较采用方差分析。结果 筛选出HILPDA基因(在2型糖尿病中上调11.35倍,腹主动脉瘤中上调6.4倍)；亚细胞定位结果预测HILPDA多分布于细胞核和线粒体中,荧光染色共定位分析证实了HILPDA在血管平滑肌细胞的细胞核和线粒体中有较高的表达；蛋白质互作网络分析证实HILPDA相关蛋白与细胞对缺氧的反应、脂质滴相关。与瘤旁动脉组织(对照组)比较,在糖尿病合并AAA患者组织中HILPDA蛋白的高表达更为显著[(47.2±5.6)% 和(71.5±4.7)%,P<0.01)]。结论 HILPDA基因对糖尿病合并腹主动脉瘤的诊断和治疗的临床研究可能具有潜在的指导价值。

Abstract:

Objective To find the differentially expressed genes (DEGs) in diabetic patients combined with abdominal aortic aneurysms (AAA) through bioinformatics analysis, then clarify their functions, and explore the roles in the pathogenesis. Methods The expression profiles of GSE21321 (diabetes mellitus) and GSE7084 (AAA) were downloaded from the Gene Expression Omnibus (GEO) database. The 2 microarray datasets were integrated to obtain DEGs by screening at P<0.05 and expression changes greater than 2 times. The subcellular localization of obtained proteins was analyzed by using Genscript database and Predictprotein platform, which were further verified with fluorescence co-localization analysis. The protein interactions of hypoxia-inducible lipid droplet-associated (HILPDA)were analyzed by the search tool for the retrival of interacting genes (STRING) database. SPSS statistics 13.0 was used for data analysis. Difference of measurement data was compared with analysis of variance. Results HILPDA was screen out, with up-regulation by 11.35 times in type 2 diabetes mellitus and by 6.4 times in AAA. The results of subcellular localization predicted that HILPDA is mostly distributed in the nucleus and mitochondria. Immunofluorescence co-localization analysis confirmed that HILPDA had a higher expression in the nucleus and the mitochondria of vascular smooth muscle cells; protein-protein interaction (PPI) network analysis confirmed that HILPDA-related proteins are related to cellular response to hypoxia and lipid droplet. HILPDA protein expression was significantly higher in the AAA tissues than the peritumor arterial tissues (control group) [(47.2±5.6)% vs (71.5±4.7)%, P<0.01].Conclusion HILPDA gene might have potential guiding value in the clinical studies concerning the diagnosis and treatment of diabetes accompanied with AAA.