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目的探讨抵抗素对内皮细胞NO生成的影响及其可能的信号机制。方法分离、培养人脐静脉内皮细胞(HUVECs),以不同浓度抵抗素(1 5、50、100 ng/ml)干预。荧光显微镜检测各组细胞中NO的生成,RT-PCR检测eNOS mRNA表达水平,Westernblot检测Akt和eNOS磷酸化水平。结果 15、50、100ng/ml抵抗素干预HUVECs 24h后,胰岛素刺激的内皮NO生成显著降低(三组分别为4.01±0.69、3.76±0.71、3.73±0.45,vs对照组P均<0.05),同时伴有内皮Akt和eNOS磷酸化水平的降低(vs对照组P均<0.05),而eNOS mRNA表达无显著改变。结论抵抗素可通过PI3K/Akt途径影响HUVECs eNOS磷酸化水平,进而调节内皮细胞NO生成,但该影响并非Akt依赖性。
Objective To investigate the effect of resistin on NO production in endothelial cells and its possible signaling mechanism. Methods Human umbilical vein endothelial cells (HUVECs) were isolated and cultured with different concentrations of resistin (15, 50 and 100 ng / ml). The production of NO was detected by fluorescence microscopy, the expression of eNOS mRNA was detected by RT-PCR and the phosphorylation of Akt and eNOS by Western blot. Results After treated with 15, 50 and 100 ng / ml of resistin for 24 h, the NO production was significantly decreased (4.01 ± 0.69, 3.76 ± 0.71 and 3.73 ± 0.45, respectively, vs control group, P <0.05) Accompanied by decreased phosphorylation of Akt and eNOS in endothelial cells (all P <0.05 vs control group), while eNOS mRNA expression did not change significantly. Conclusion The resistin can affect the phosphorylation of eNOS in HUVECs through the PI3K / Akt pathway and then regulate the NO production in endothelial cells, but the effect is not Akt-dependent.