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目的研究抵抗素对人脐静脉内皮细胞(HUVEC)一氧化氮生成的影响及其信号机制。方法不同浓度人抵抗素(0～100 ng/ml)干预HUVEC 24h。DAF-2DA染色,激光共聚焦显微镜观察各组一氧化氮生成改变,Westem-Blot检测各组eNOS、AMPK磷酸化水平改变。结果 15、50、100 ng/ml抵抗素干预HUJVEC 24 h后,3组的一氧化氮生成分别为4.01±0.69、3.76±0.71、3.73±0.45,与对照组一氧化±氮生成(4.89 ±0.58)相比显著降低(P<0.05);50 ng/ml组添加AMPK特异激动剂AICAR后,一氧化氮生成(5.08±0.70)显著升高(P<0.01)。各抵抗素干预组的AMPK和eNOS磷酸化水平均明显降低;而添加AMPK特异激动剂AICAR后,伴随AMPK的激活,eNOS磷酸化水平也显著增高(P<0.05)。结论抵抗素在内皮细胞可通过对AMPK的抑制进而导致eNOS失调,降低内皮一氧化氮的生成。
Objective To study the effect of resistin on nitric oxide production in human umbilical vein endothelial cells (HUVEC) and its signaling mechanism. Methods Different concentrations of human resistin (0 ~ 100 ng / ml) interfered HUVEC for 24h. DAF-2DA staining, laser confocal microscopy changes in nitric oxide production in each group, Westem-Blot test eNOS, AMPK phosphorylation levels change. Results The NO production of HUJVEC was 4.01 ± 0.69, 3.76 ± 0.71 and 3.73 ± 0.45 respectively after intervention with HUFVEC for 24 h, and was significantly higher than that of the control group (4.89 ± 0.58% ) (P <0.05). After AICAR was added to 50 ng / ml group, NO production was significantly increased (5.08 ± 0.70) (P <0.01). The phosphorylation levels of AMPK and eNOS in each resistin group were significantly decreased. However, the phosphorylation of eNOS was also significantly increased with the AMPK agonist AICAR (P <0.05). Conclusion The resistance of resistin to endothelial cells may lead to the deregulation of eNOS and the decrease of endothelial nitric oxide production.