高尿酸通过miR-663下调转化生长因子β_1抑制内皮细胞迁移  

High uric acid concentration inhibits endothelial cell migration by down-regulating TGF-β_1 expression via miRNA-663

作　　者：于善栋 洪权[1] 傅博[1] 陈香美[1] 吴镝[1] 

机构地区：[1]解放军总医院肾脏病科暨肾脏疾病国家重点实验室,北京100853

出　　处：《解放军医学院学报》2014年第7期733-737,共5页Academic Journal of Chinese PLA Medical School

基　　金：国家自然科学基金面上项目(31170810;81102673);北京科技新星计划(Z121107002512078)~~

摘　　要：目的探讨高尿酸通过miR-663影响内皮细胞迁移功能的机制。方法培养人脐静脉内皮细胞系(EA.hy926),用600μmol/L尿酸孵育48 h,实时定量PCR检测miR-663的水平,并检测高尿酸患者血清中miR-663水平;利用双荧光素酶试验预测并验证miR-663的靶基因;检测高尿酸条件下内皮细胞中转化生长因子β1(transforming growth fator beta 1,TGF-β1)的表达水平,利用miR-663抑制物、TGFB1 siRNA转染及划痕实验观察高尿酸如何通过miR-663及其靶基因影响内皮细胞的迁移功能。结果高尿酸培养条件下内皮细胞中miR-663表达水平明显升高,高尿酸血症患者血清中miR-663的水平也高于正常人;双荧光素酶试验结果表明TGFB1的翻译水平受miR-663直接调控;高尿酸能明显抑制细胞的迁移能力,而高尿酸条件下TGF-β1的表达水平也明显下降,转染miR-663抑制物后,TGF-β1表达水平升高,内皮细胞迁移能力也明显改善,但是利用siRNA抑制TGF-β1的表达后,miR-663抑制物不能再促进内皮细胞的迁移。结论高尿酸通过miRNA-663下调TGF-β1而抑制细胞迁移。Objective To study the mechanism of high uric acid （UA） concentration underlying endothelial cell migration via miRNA-663. Methods EA.hy926 cells were incubated in 600 μ mol/L uric acid for 48 h. Serum miRNA-663 level in patients with high UA concentration was measured by RT-PCR. The miRNA-663 target gene was identified by dual luciferase assay. TGF- β1 expression level in endothelial cells was measured. Effect of high UA concentration on endothelial cell migration via miRNA-663 was detected by miR-663 inhibitor and TGF-β1 siRNA transfection and scratch test, respectively. Results The expression level of miRNA-663 was significantly higher in endothelial ceils after cultured with high UA concentration than before cultured with high UA concentration. The serum miRNA-663 level was significantly higher in hyperuricemia patients than in normal subjects. Dual luciferase assay showed that miRNA-663 directly regulated the TGFB 1 translation level. Scratch test revealed that high UA concentration significantly inhibited the endothelial cell migration and down-regulated the TGF- β1 expression level. The TGF- β1 expression level elevated and the endothelial cell migration increased after the miRNA-663 inhibitors were transfected. However, the miRNA-663 inhibitors could not promote endothelial cell migration when the TGF- β1 expression was inhibited by siRNA. Conclusion High UA concentration inhibits endothelial cell migration by down-regulating TGF- β1 expression via miRNA-663

关 键 词：高尿酸 内皮细胞迁移 miRNA-663 转化生长因子Β1 

分 类 号：R543[医药卫生—心血管疾病]