Ding Li, Zhang Jin, Zhang Shao-Wei, Zhao Wen-Zhou
Department of Endocrinology, First Affiliated Hospital, China Medical University, Shenyang, China.
Zhonghua Yi Xue Za Zhi. 2012 Nov 13;92(42):3008-11. doi: 10.3760/cma.j.issn.0376-2491.2012.42.016.
To explore the effects of glucagon-like peptide 1 (GLP-1(9-36)) on endothelial nitric oxide synthase (eNOS) in human umbilical vein endothelial cells (HUVEC).
HUVEC cultured under the conditions of normal glucose (5.5 mmol/L) or high glucose (16.8 mmol/L) were incubated with 5-5000 pmol/L GLP-1(9-36). NO production was assayed by the nitrate reductase method. The eNOS activities were detected by NOS assay kit, p-eNOS (ser-1177) level and total eNOS protein level by Western blot and eNOS mRNA level by real-time reverse transcription-polymerase chain reaction (RT-PCR).
Under normal glucose condition, NO productions from HUVEC of 50 - 5000 pmol/L GLP-1(9-36) groups were significantly higher than the control ((41.6 ± 8.1) µmol/L vs (22.2 ± 2.6) µmol/L, P < 0.05). So were eNOS activities in cells (1.76 ± 0.12 vs 1.00 ± 0.00, P < 0.05). Compared with the control group, the levels of eNOS phosphorylation at ser-1177, mRNA and total protein were significantly elevated in the 5000 pmol/L GLP-1(9-36) group. Under high glucose condition, GLP-1(9-36) retained all the above effects.
GLP-1 (9-36) can increase NO release, eNOS activity and expression in HUVEC. This may be one of the underlying mechanisms for the protective effects of GLP-1(9-36) on cardiovascular system.
探讨胰高血糖素样肽1(GLP-1(9-36))对人脐静脉内皮细胞(HUVEC)中内皮型一氧化氮合酶(eNOS)的影响。
将在正常葡萄糖(5.5 mmol/L)或高葡萄糖(16.8 mmol/L)条件下培养的HUVEC与5 - 5000 pmol/L的GLP-1(9-36)一起孵育。采用硝酸还原酶法检测一氧化氮(NO)生成量。用一氧化氮合酶检测试剂盒检测eNOS活性,通过蛋白质免疫印迹法检测p-eNOS(丝氨酸1177位点)水平和总eNOS蛋白水平,通过实时逆转录-聚合酶链反应(RT-PCR)检测eNOS mRNA水平。
在正常葡萄糖条件下,50 - 5000 pmol/L GLP-1(9-36)组HUVEC产生的NO量显著高于对照组((41.6 ± 8.1) μmol/L对(22.2 ± 2.6) μmol/L,P < 0.05)。细胞中的eNOS活性也显著高于对照组(1.76 ± 0.12对1.00 ± 0.00,P < 0.05)。与对照组相比,5000 pmol/L GLP-1(9-36)组中丝氨酸1177位点的eNOS磷酸化水平、mRNA和总蛋白水平均显著升高。在高葡萄糖条件下,GLP-1(9-36)仍具有上述所有作用。
GLP-1(9-36)可增加HUVEC中NO的释放、eNOS活性及表达。这可能是GLP-1(9-36)对心血管系统起保护作用的潜在机制之一。