摘要
目的探讨在高糖培养环境下西格列汀对人主动脉内皮细胞(HAEC)内皮素1及一氧化氮合酶的影响及其可能机制。方法高糖环境下(25 mmol/L葡萄糖)培养人主动脉内皮细胞,并分别给予0、5、10及20μmol/L西格列汀干预,检测HAEC内皮型一氧化氮合酶(e NOS)、内皮素1(ET-1)、诱导型一氧化氮合酶(i NOS)以及核因子κB p65(NF-κB p65)的mRNA和蛋白表达水平;应用肿瘤坏死因子α(TNF-α)处理西格列汀干预后的HAEC,观察e NOS、ET-1、i NOS、NF-κB p65的mRNA和蛋白表达水平的变化。结果与普通培养环境(7 mmol/L葡萄糖)相比,高糖环境下HAEC的e NOS mRNA和蛋白表达降低,ET-1、i NOS mRNA和蛋白表达及p-NF-κB p65蛋白表达增高(P<0.05)。与0μmol/L组相比,20μmol/L西格列汀组HAEC的e NOS mRNA、蛋白表达上调,ET-1、i NOS mRNA和蛋白表达以及p-NF-κB p65蛋白表达下调(P<0.05)。与单独使用西格列汀组相比,TNF-α联合西格列汀组HAEC的e NOS mRNA和蛋白表达下调,ET-1、i NOS mRNA和蛋白表达及p-NF-κB p65蛋白表达上调(P<0.05)。结论西格列汀在转录和翻译水平上增加e NOS表达、降低ET-1、i NOS的表达,该作用通过其抑制p-NF-κB p65表达而实现。提示西格列汀可能改善内皮功能,从而预防糖尿病动脉粥样硬化的形成。
Aim To explore the effect of sitagliptin on the expression of endothelin-1（ ET-1） and endothelial nitric oxide synthase（ e NOS） in human aortic endothelial cells（ HAEC） and its underlying mechanism in high glucose environment. Methods HAEC were cultured in high glucose environment（ 25 mmol / L）,and treated with different concentrations of sitagliptin（ 0,5,10 and 20 μmol / L,respectively）. The mRNA and protein expressions of e NOS,ET-1,i NOS and phosphate nuclear factor-kappa B p65（ p-NF-κB p65） were measured. The measurements for e NOS,ET-1,i NOS,NF-κB p65 on mRNA and protein levels in HAEC were evaluated after incubation with tumor necrosis factor-α（ TNF-α） and sitagliptin. Results Compared with normal medium（ glucose concentrations for 7 mmol/L）,both the mRNA and the protein expression of e NOS in HAEC significantly decreased in high glucose medium,while those of ET-1,i NOS and p-NF-κB p65 protein significantly increased（ P〈0. 05）. Compared with 0 μmol / L sitagliptin,20 μmol / L sitagliptin significantly increased mRNA and protein expressions of e NOS,while decreased those of ET-1,i NOS and p-NF-κB p65 protein（ P〈0. 05）. Compared with sitagliptin alone treated HAEC,both the mRNA and the protein expressions of e NOS significantly decreased in HAEC treated with TNF-α and sitagliptin,while those of ET-1,i NOS and p-NF-κB p65 protein expressions significantly increased（ P〈0. 05）. Conclusions Sitagliptin enhances e NOS,represses ET-1,i NOS expressions at the level of transcription and translation through inhibiting NF-κB p65 phosphorylation in HAEC in high glucose environment. This may contribute to the improvement of endothelial function and prevention of subsequent atherogenesis.
出处
《中国动脉硬化杂志》
CAS
北大核心
2016年第2期114-118,共5页
Chinese Journal of Arteriosclerosis
基金
国家自然科学基金青年科学基金培养计划项目(2013KJ25)
安徽省自然科学基金面上项目(1508085MH178)
