罗格列酮对人脐静脉内皮细胞NO和PI3K/PKB/eNOS信号通路的影响  

Effect of rosiglitazone on NO and eNOS via PI3K/PKB signal pathways in cultured human umbilical vein endothelial cells

作　　者：吴静[1] 雷闽湘[1] 谢小云[1] 冯湘玲[2] 

机构地区：[1]中南大学湘雅医院内分泌科,长沙410008 [2]中南大学肿瘤研究所,长沙410078

出　　处：《中南大学学报（医学版）》2007年第5期824-830,共7页Journal of Central South University ：Medical Science

基　　金：湖南省科技厅基金(06sk3023)~~

摘　　要：目的:观察罗格列酮对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)一氧化氮(nitric oxide,NO)浓度和内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)、磷脂酰肌醇-3激酶(phosphatidylinositol3-kinase,PI3K)和蛋白激酶B(protein kinase B,PKB)表达的影响,探讨罗格列酮改善内皮功能的信号转导机制。方法:首先观察罗格列酮对内皮细胞NO影响的时效和量效关系,然后加用eNOS和PKB信号阻断剂进行干预。用Griess重氮化反应法检测NO浓度,RT-PCR方法检测PI3K,PKB及eNOS mRNA的表达,Western免疫印迹方法检测PKB,eNOS总蛋白和PKB丝氨酸473(PKB-Ser473),eNOS丝氨酸1177(eNOS-Ser1177)的磷酸化表达。结果:罗格列酮呈剂量和时间依赖性地升高内皮细胞NO浓度,不同浓度的罗格列酮培养内皮细胞均能升高PI3K mRNA表达和PKB-Ser473,eNOS-Ser1177磷酸化,但对PKB和eNOS表达均无影响;eNOS阻断剂L-NAME能完全阻断罗格列酮培养的内皮细胞NO浓度的升高,PI3K阻断剂LY294002能阻断罗格列酮诱导的NO产生和PKB,eNOS的磷酸化。结论:罗格列酮能通过激活PI3K/PKB/eNOS信号通路而增强内皮细胞NO的浓度,改善内皮功能。Objective To observe the effect of rosiglitazone on the production of nitric oxide （NO） and the expression of phosphatidylinositol 3-kinase （PI3K）/protein kinase B （PKB） /the endothelial nitric oxide synthase （eNOS） in cultured human umbilical vein endothelial cells （HU-VECs ） , and to investigate the mechanism of signal transduction of rosiglitazone in improving the en-dothelial function. Methods HUVECs were treated with various concentrations of rosiglitazone. The NO level was measured using Griess Reaction in cell culture supernatants ; the expressions of PI3 K-,PKB- and eNOS mRNA were measured using RT-PCR; and the expresstions of PKB, eNOS, and phosphorylation of PKB-Ser473, eNOS-Ser1177 were measured using Western Blot. Results Rosiglitazone increased the endothelial NO production in a dose- and time-dependent manner in cul-tured HUVECs, and also increased the expression of PI3K mRNA and the phosphorylation of PKB-Ser473 and eNOS-Ser1177 in a concentration-dependent manner, with no alteration in the expression of PKB and eNOS in cultured HUVECs. N （w）-nitro-L- arginine methyl ester （L-NAME,eNOS synthase inhibitor） blocked the rosiglitazone-induced NO formation; LY294002 （a PI3K inhibitor） prevented the NO production; and the phosphorylation of eNOS and PKB was induced by rosiglitazone. Conclusion Treatment with rosiglitazone can increase the NO prouducion and improve the endothelial function through up-regulating the PI3K/PKB/eNOS signal pathways in cultured HU-VECs.

关 键 词：罗格列酮 一氧化氮 内皮型一氧化氮合酶 磷脂酰肌醇-3激酶 蛋白激酶B 

分 类 号：R96[医药卫生—药理学]