糖化低密度脂蛋白对人脐静脉内皮细胞影响的数字基因表达谱分析  

Digital gene expression profile analysis of glycated low-density lipoprotein impacted on human umbilical vein endothelial cells

作　　者：张冰雨[1] 曾瑞翔 雷涛[2] 

机构地区：[1] 同济大学附属同济医院内分泌科,上海200065 [2] 上海中医药大学附属普陀医院内分泌科

出　　处：《中华内分泌代谢杂志》2016年第11期940-945,共6页Chinese Journal of Endocrinology and Metabolism

基　　金：基金项目：863科研专项基金（2014AA022304）;上海市卫生局科研课题（20124256）

摘　　要：目的：从基因水平探讨糖化低密度脂蛋白（ LDL ）对血管内皮细胞的作用。方法用20 mmol/L葡萄糖将2 mg/ml LDL在37℃无菌抗氧化条件下孵育4周完成糖化,然后用完全培养基及0.1 mmol/L糖化LDL（gly-LDL）处理人脐静脉内皮细胞（HUVECs）24 h,设普通LDL（n-LDL）组和空白对照组,提取总RNA,进行数字表达谱分析（ DGE）,并选取部分基因进行实时定量PCR（ q-PCR）验证。结果对照组（ C）与普通LDL组（ N）相比有2287个差异基因；C与糖化LDL组（ G）相比有4721个差异基因；N与G相比有442个差异基因；在C与G之间及N与G之间均存在显著性差异,有327个基因在C与G、N和G组间存在显著差异。包括221个下调差异基因和106个上调差异基因。 KEGG富集分析显示C与G的差异基因可映射到199条信号通路中,其中显著性富集（P〈0.05）的有71条,C与N的差异基因可映射到193条信号通路中,显著性富集的有70条,N与G的差异基因可映射到148条信号通路中,显著性富集的有41条。 q-PCR结果与DGE结果相符。结论 Gly-LDL可能影响血管内皮细胞的迁移,同时也影响炎症相关因子的表达,另外gly-LDL可能与糖尿病并发肿瘤相关。Objective To study the effect of glycated low-density lipoprotein（ gly-LDL） on human vascular endothelial cell on gene level. Methods Under the condition of asepsis and with antioxidants, incubated 2 mg/ml of LDL with 20 mmol/L of glucose in sealed tubes overlaid with nitrogen at 37℃ for 4 weeks. The endothelial cells were cultured with complete medium and 0. 1 mg/ml glycated low-density lipoprotein for 24 h in the incubator, while the control group were cultured with LDL without chemical modification or PBS for 24 h. Total RNA was extracted for the analysis of digital gene expression spectrum （ DGEs ） . Finally, several genes were selected for the real time quantitative PCR（ q-PCR） experiment. Results There are 2 287 different expressed genes between the control group （C） and normal LDL group（N）, 4 721 different expressed genes between C and glycated LDL group（G）, 442 different genes between N and G. There were 327 genes having significant difference between C and G, N and G, but no significant difference between C and N, including 221 down-regulated and 106 up-regulated differential expressed genes. KEGG enrichment analysis shows that the different genes between C and G could be mapped to 199 pathways, there were 71 pathways having significant values（P〈0. 05）, the different genes between C and N can be mapped to 193 pathways, there were 70 pathways having significant value, the different genes between N and G can be mapped to 148 pathways, there were 41 pathways having significant value. The results of q-PCR were conformed to the results of DGEs. Conclusion Gly-LDL may affect the migration of endothelial cells, and may also influence the expression of inflammation-related factors. Besides, gly-LDL may be associated with diabetes accompanying tumors.

关 键 词：糖尿病 血管病变 糖化低密度脂蛋白 人脐静脉内皮细胞 数字基因表达谱 

分 类 号：R587.2[医药卫生—内分泌]