利拉鲁肽对人脐静脉内皮细胞一氧化氮合酶及胰岛素受体底物-1表达的影响

The effect of liraglutide on the expression of eNOS and IRS-1 in human umbilical vein endothelial cells

目的探讨胰升血糖素样肽-1(GLP-1)类似物利拉鲁肽对体外培养的人脐静脉内皮细胞(HUVECs)胰岛素受体底物-1(IRS-1)、一氧化氮合酶(eNOS)和磷酸化eNOS(p-eNOS)表达的影响。方法以含有5.5mmol/L葡萄糖的DMEM培养基体外培养HUVECs,将上述细胞分为10组,各组中均加入3nmol/L的利拉鲁肽注射液干预,分别于第0、10、15、30、45、60、90、120、150、180min提取细胞,以Western blot检测各组细胞IRS-1、eNOS及p-eNOS表达情况。结果利拉鲁肽干预HUVECs不同时间后,各组细胞表达IRS-1和p-eNOS有所不同,0、10、15min细胞p-eNOS表达比较,差异无统计学意义(P>0.05)。从第30min开始,p-eNOS表达逐渐增加,至第150min达峰,随之p-eNOS表达量开始下降(P<0.05);IRS-1表达趋势与p-eNOS相似,也具有利拉鲁肽时间依赖性,且作用高峰亦为150min(P<0.05);而各时间点eNOS表达比较,差异均无统计学意义(P>0.05)。结论 GLP-1类似物利拉鲁肽呈时间依赖性的促进正糖培养的HUVECs IRS-1及p-eNOS的表达,其作用高峰为第150 min,而对eNOS的表达未观察到促进作用。

Objective To explore the effects of glucagon like peptide-1（GLP-1）analogue liraglutide on the expression of endothelial nitric oxide synthase（eNOS）,phosphorylated endothelial nitric oxide synthase（p-eNOS）and insulin receptor substrates-1（IRS-1）in human umbilical vein endothelial cells（HUVECs）in vitro. Methods HUVECs were cultured in DMEM containing 5.5mmol/L glucose,and then divided into 10 groups.HUVECs of each group were incubated with liraglutide（3nmol/L）.A fter incubation,cells were extracted in the 0th,10 th,15th,30 th,45th,60 th,90th,120 th,150th and 180 th minutes＇ respectively.The expression levels of eNOS,p-eNOS and IRS-1at each point were measured by Western blot. Results There were no significant difference of p-eNOS levels in 0th,10 th,15th minutes（P〉0.05）.However,p-eNOS levels were observed to increase in 30 th minute and reach a maximum in150 th minute（P〈0.05）,and then present a descent trend.IRS-1activity followed a similar trend of peNOS in a time-dependent manner of liraglutide.The levels of eNOS had no difference at different points（P〈0.05）. Conclution Liraglutide showed a time-dependent manner to promote the expressions of peNOS and IRS-1in euglycemic cultured HUVECs with peak effect time in 150 th minute.However,there was no effect of liralutide on the expressions of eNOS.