摘要
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 HUVECs NO production and on the Akt signaling. Methods HUVECs were isolated and cultured for experiment. Endothelial cells were incubated with recombinant human resistin （15, 50, 100ng/ml） for 24 hours and stimulated by insulin for 20 min. NO production of HUVECs was detected by fluorescence microscope. The eNOS mRNA expression level and eNOS and Akt phoshporylation levels were measured by RT-PCR and Western-blot. Results Interventions by resistin for 24h resulted in the decrease of insulin-stimulated NO production （Convs resistin of 15, 50, 100ng/ml group：4.89±0.58 versus 4.01±0.69, 3.76±0. 71, 3. 734±0.45 respectively, P 〈 0. 05）. At the same time, Akt and eNOS phosphorylation levels were decreased obviously, hut there was no significant change in eNOS mRNA expression level. Conclusions Resistin may inhibit the NO production of HUVECs through the regulation of eNOS phosphorylation and may be independent on PI3K/Akt signaling of insulin.
出处
《中国糖尿病杂志》
CAS
CSCD
北大核心
2009年第7期522-524,528,共4页
Chinese Journal of Diabetes
基金
国家自然科学基金资助项目(编号30570886)
关键词
抵抗素
脐静脉内皮细胞
一氧化氮
蛋白激酶B
Resistin
Umbilical vein endothelial cells
Nitric oxide
Protein kinase B
