·糖尿病基础研究·

李志臻 李芳萍 叶建红 周嘉 李焱 严励 傅祖植

基金项目：国家自然科学基金资助项目(30570886)

作者单位:510120广州，中山大学附属第二医院内分泌科(第一作者现在郑州大学第一附属医院内分泌科工作)

【摘要】 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，Western-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HUVEC 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。

【关键词】 抵抗素,内皮细胞,AMPK,一氧化氮合酶

doi:10.3969/j.issn.1006-6187.2011.07.017

The effect of resistin on HUVECs NO production and the correlated AMPK pathway

LI Zhi-zhen,LI Fang-ping,YE Jian-hong, et al.

Department of Endocrinology, Second Affiliated Hospital, SUN Yat-sen University, Guangzhou 510120, China

【Abstract】 Objective To investigate the effects of resistin on nitric oxide(NO) production in human umbilical vein endothelial cells(HUVECs). Method sHUVECs were incubated with recombinant human resistin (0~100ng/ml) for 24 hours. The NO production was detected by DAF-2DA labeling. AMPK and eNOS phosphorylation levels were measured by Western blot. Result sThe NO production of HUVECs treated by 15,50,100 ng/ml resistin were 4.01±0.69,3.76±0.71,3.73±0.45 respectively, they were significantly lower than control group(4.89±0.58，P<0.05). AICAR, the specific activator of AMPK, can stimulate NO production significantly. With the decreasing of NO production of each rsistin treatment group, the eNOS and AMPK phosphorylation levels decreased significantly. And while accompanied by AICAR-induced AMPK activation, eNOS phosphorylation level was increased obviously. Conclusion Resistin exerts an inhibitory effect on the NO production in HUVECs, and induces the dysregulation of endothelial eNOS via the AMPK pathway.

【Key words】 Resistin; Endothelium; AMPK;NOS