视黄醇类X受体激动剂抑制高糖诱导内皮细胞氧化应激  

Retinoid X receptor Agonists Inhibit High-Glucose-Induced Oxidative Stress in Human Endothelial Cells

作　　者：柴大军[1] 林金秀[1] 许昌声[1] 沈玲红[2] 王彬尧[2] 何奔[2] 

机构地区：[1]福建医科大学附属第一医院心血管内科,福建福州350005 [2]上海交通大学医学院附属仁济医院心血管内科,上海200127

出　　处：《中华高血压杂志》2009年第11期1010-1014,共5页Chinese Journal of Hypertension

基　　金：福建省科技三项基金项目(2006F5039);国家自然基金(30600242)资助

摘　　要：背景还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶是在高糖环境下内皮细胞活性氧离子(ROS)产生的重要来源,视黄醇类X受体(RXR)能够抑制高糖诱导下内皮细胞ROS的生成,但对NADPH氧化酶的表达水平的影响尚不清楚。目的探讨高糖干预下血管内皮细胞NADPH氧化酶NOX4、gp91phox和p22phox亚基表达水平的变化及RXR激动剂的影响。方法体外培养人脐静脉血管内皮细胞(HUVECs),以33mmol/L葡萄糖干预,模拟糖尿病患者体内环境,通过流式细胞学方法检测细胞内的ROS水平,采用干扰RNA(siRNA)技术检测NADPH氧化酶NOX4、gp91phox和p22phox亚基在高糖诱导下内皮细胞产生氧化应激反应中的作用,分别应用实时荧光定量PCR和免疫印迹杂交的方法检测NADPH氧化酶各亚基的表达水平。结果高糖处理后HUVECs内ROS的水平显著增加(6.58±0.24对2.47±0.12,P<0.05)。siRNA检测结果显示NOX4、gp91phox和p22phox亚基参与了高糖诱导下HUVECs内ROS的生成过程。高糖处理显著上调NADPH氧化酶NOX4、gp91phox和p22phox亚基的mRNA水平(NOX4:0.390±0.020比0.110±0.006,P<0.05;gp91phox:0.480±0.040比0.090±0.005,P<0.05;p22phox:0.025±0.002比0.005±0.001,P<0.05)和蛋白水平。RXR激动剂9-cis-RA和SR11237抑制高糖诱导下ROS生成,以及NADPH氧化酶NOX4、gp91phox和p22phox各亚基mRNA和蛋白水平的表达。结论RXR激动剂通过下调高糖诱导下内皮细胞NADPH氧化酶的表达,对抗高糖诱导内皮细胞产生的氧化应激反应。Background Nicoti.namide adenine dinueleotide phosphate （NADPH） oxidase plays a key role in high-glucose-induced oxidative stress in human endothelial cells, retinoid X receptor （RXR） agonists can decrease high glucose-induced reactive oxygen species（ROS） production. However, the effects of RXR ligands on individual subunit of NADPH oxidase had not been well delineated. Objective To investigate the role of NOX4, gp91phox and p22phox subunits of NADPH oxidase in high glucose-induced oxidative stress in human endothelial cells and whether RXR agonists was able to attenuate the production of ROS mediating inhibited expression of subunits of NADPH oxidase in vitro. Methods Human umbilical vein endothelial cells （HUVECs） were isolated from healthy umbilical cords and cultured. Antisense transfection was obtained to reveal the role of subunits of NADPH oxidase in highglucoseqnduced oxidative stress. Reverse transcription and real time PCR and immunoblotting were performed to determine the expression of the subunits of NADPH oxidase and NOX4. ROS production was detected using flow cytometry. Results ROS level was significantly increased in high-glucose-induced HUVECs （6.58±0.24 vs 2.47± 0.12, P〈0.05）. High-glucose-induced ROS production in HUVECs was mainly mediated through its activation of the NOX4, gp91phox and p22phox subunits of NADPH oxidase. The mRNA levels of NOX4, gp91phox, and p22phox subunits of NADPH oxidase were significantly up-regulated by high glucose （NOX4： 0. 390±0. 020 vs 0. 110± 0.006, P〈0.05; gp91phox： 0.480±0.040 VS 0.090±0.005, P〈0.05; p22phox ： 0.025±0.002 vs 0.005±0.001, P〈0.05）. High glucose has similar effect on protein level of these subunits. Treatment of endothelial cells with RXR agonists 9-cis-RA and SRl1237 resulted in significant inhibition of ROS production and attenuate the expression of NOX4, gp9tphox , and p22phox induced by high glucose levels.. Conclusion RXR agonists inhibit high-glucose-induced ROS production by repressing the expres

关 键 词：视黄醇类X受体 高糖 氧化应激 还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶 

分 类 号：R587.1[医药卫生—内分泌]