【作者】 温志桥；

【导师】 金红；

【作者基本信息】 汕头大学 ， 心血管内科， 2010， 硕士

【摘要】 背景:血管内皮细胞功能紊乱是代谢综合征(metabolic syndrome, MS)的一个重要特征,而抵抗素(resistin, Re)为新近发现的脂肪细胞因子,在代谢综合征发生、发展过程中起重要的调控作用,抵抗素可能促进血管内皮细胞功能紊乱。目前已证实一氧化氮(nitric oxide, NO)及环磷腺苷(cyclic adenosine monophosphate, cAMP)在维持正常的血管内皮功能中起重要作用。目的:本研究主要观察抵抗素对人脐静脉内皮细胞(human umbilical vein endothelial cells, HUVECs)内NO的分泌及cAMP水平的影响,探讨其对血管内皮功能调节的作用机制。方法:(1)体外培养HUVECs,取4-6代细胞用于实验。培养的HUVECs接种于96孔板,生长至80%以上融合状态,将细胞分为3组:Ⅰ组为单纯培养基对照组,Ⅱ组为50ng/ml抵抗素组,Ⅲ组为20ng/ml肿瘤坏死因子-α(tumor necrosis factor-α, TNF-α)组。加入上述试剂处理12小时后,取上清液离心去除细胞碎片,硝酸还原酶法测定NO含量。(2)体外培养HUVECs,取4-6代细胞用于实验。培养的HUVECs接种于细胞培养瓶中,生长至80%以上融合状态,分组同上。加入上述试剂处理培养6小时后,提取蛋白,ELISA法测定cAMP含量。结果:(1)与单纯培养基对照组相比,抵抗素组及TNF-α组均能轻度抑制HUVECs内NO的分泌,但无统计学意义(P>0.05)。(2)与单纯培养基对照组相比,经抵抗素及TNF-α培养6小时后,HUVECs内cAMP的水平均明显下降(P<0.05)。结论:抵抗素对HUVECs内NO的分泌无明显影响,但能明显降低cAMP水平,提示抵抗素可能通过抑制cAMP合成而引起血管内皮功能障碍。更多还原

【Abstract】 Background:Vascular endothelial cells dysfunction is a main trait of metabolic syndrome (MS), and resistin is a recently discovered adipokinine, which plays an important role in the pathophysiological development of MS. Resistin may cast adverse effects on endothelial cell dysfunction. Nitric oxide (NO) and cyclic adenosine monophosphate (cAMP) have been found to play important roles in maintenance of vascular endothelial function.Objective: In this study we evaluated the effects of resistin on NO and cAMP levels in cultured human umbilical vein endothelial cells (HUVECs) to explore the potential mechanism of resistin on vascular endothelial function mediation.Methods: (1) Cultured HUVECs within passage 4-6 were inoculated in 96-well plate to reach 80 percent of confluence to be divided into 3 treatment groups:Ⅰmedium only control group,Ⅱ50ng/ml resistin incubation group,Ⅲ20ng/ml tumor necrosis factor-α(TNF-α) incubation group for 12h, and the supernatant obtained from the cultured HUVECs were collected, centrifuged to remove cell debris, then measured with the method of Nitrate reductase.(2) Cultured HUVECs within passage 4-6 were inoculated in Cell Culture Flasks to reach 80 percent of confluence to be divided into 3 treatment groups:Ⅰmedium only control group,Ⅱ50ng/ml resistin incubation group,Ⅲ20ng/ml TNF-αincubation group for 6h. Extracted and then measured the cAMP levels with Enzyme Linked Immunosorbent Assay kits (ELISA).Result: (1) Compared with control group, neither 50ng/ml resistin nor 20ng/ml TNF-αdecreased NO levels significantly in cultured HUVECs (p>0.05).(2) Compared with that of the control group, cAMP levels were significantly lower in resistin or TNF-αtreatment cells (p<0.05).Conclusion: Resistin has no effect on the secretion of NO from HUVECs, but decreased the cAMP level significantly, suggesting that resistin may cause endothelial dysfunction, partly, by reducing cAMP concentration.更多还原