High glucose inhibits expression of inducible and constitutive nitric oxide synthase in bovine aortic endothelial cells  

高糖抑制牛主动脉内皮细胞诱导型和结构型一氧化氮合酶的表达(英文)

作　　者：郭迅[1] 谌立伟[1] 刘文兰[1] 郭兆贵[1] 

机构地区：[1]湖南医科大学分子药理研究室

出　　处：《Acta Pharmacologica Sinica》2000年第4期325-328,共4页中国药理学报（英文版）

摘　　要：AIM: To investigate the effects of high glucose on the expression of nitric-oxide synthase (NOS) in cultured bovine aortic endothelial cells (BAEC). METHODS: BAEC were cultured and passaged in normal glucose (NG) 5.5 mmol·L-1, high glucose (HG) 25 mmol· L -1, or high osmolarity (glucose 5.5 mmol · L -1 + man-nitol 19.5 mmol ·L-1, Mann-BAEC), lipopolysacchar-ides (LPS)-induced nitric oxide (NO) production was assessed by Griess reaction. The expression of inducible NOS (iNOS) and constitutive NOS (ecNOS) was determined by Western blot. RESULTS: At a concentration range from 0.5 to 2 mg·L-1, LPS stimulated NO production in NG-BAEC in a concentration-dependent manner. NO production reached the peak level at LPS 1 mg ·L-1. HG inhibited NO production, when compared with NG- and Mann-BAEC (nitrite μmol · L-1: HG-BAEC 43±8, vs NG-BAEC 71 ±11, Mann-BAEC 70± 9, n = 4 experiments, P < 0.01). iNOS expression was decreased by 39.9 % and 39.3 %, and ecNOS by 28 % and 24 % respectively in HG-BAEC, when compared with NG- or Mann-BAEC. However, no marked difference was observed in the LPS-induced NO production and the expression of iNOS and ecNOS between NG-and Mann-BAEC. CONCLUSIONS: Inhibition of BAEC NO production by HG was mainly due to a decreased expression of NOS protein.目的:研究高糖对新生小牛主动脉内皮细胞(BAEC)一氧化氮合酶(NOS)表达的影响。方法:BAEC培养并传代于含正常葡萄糖(5.5mmol·L^(-1),NGBAEC),高糖(25 mmol·L^(-1),HG-BAEC)或高渗(葡萄糖5.5+甘露醇19.5 mmol·L^(-1),Mann-BAEC)的无酚红M1640培养基.Griess反应检测脂多糖(LPS)诱导的一氧化氮(NO)产生.Westem blot法检测结构型NOS(ecNOS)及诱导型NOS(iNOS)表达。结果:LPS(0.25-2 mg·L^(-1))剂量依赖性刺激BAEC产生NO,并在LPS 1 mg·L^(-1)达峰值。高糖显著抑制LPS诱导的NO产生(亚硝酸μmol·L^(-1):HG-BAEC 43±8,vs NG-BAEC 71±11,Mann-BAEC 70±9,n=4,P<0.01)。同样,与NG-和Mann-BAEC相比,HG-BAEC iNOS表达下降39.9％和39.3％,ecNOS表达下降28％和24％,而NG-与Mann-BAEC之间,LPS诱导的NO产量和iNOS和ecNOS的表达无差别。结论:高糖抑制BAEC NO的释放,与NOS的低表达有关。

关 键 词：nitric-oxide synthase glucose lipopo- lysaccharides Western blotting vascular endothelium cultured cells 

分 类 号：R363[医药卫生—病理学]