论文部分内容阅读
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。
Objective: To investigate the effect of high glucose on the expression of nitric oxide synthase (NOS) in newborn calf aortic endothelial cells (BAEC). Methods: BAEC was cultured and passaged on a medium containing normal glucose (5.5 mmol·L -1, NGBAEC), high glucose (25 mmol·L -1, HG-BAEC) or hypertonic (glucose 5.5 + mannitol 19.5 mmol·L -1 Mann-BAEC), and the production of nitric oxide (NO) induced by lipopolysaccharide (LPS) was measured by Griess reaction.Western blotting was used to detect the expression of structural NOS (ecNOS) And inducible NOS (iNOS) expression. RESULTS: LPS (0.25-2 mg · L -1) produced NO at a dose-dependent manner and peaked at 1 mg · L -1 of LPS. Hg-BAEC 43 ± 8, vs NG-BAEC 71 ± 11, Mann-BAEC 70 ± 9, n = 4, P <0.01 ). Similarly, expression of iNOS decreased by 39.9% and 39.3% in HG-BAEC and decreased by 28% and 24%, respectively, compared with NG- and Mann-BAEC. However, LPS-induced NO production and iNOS There was no difference in the expression of ecNOS. Conclusion: High glucose inhibits the release of BAEC NO, which is related to the low expression of NOS.