促甲状腺激素对人微血管内皮细胞一氧化氮合酶的影响及其机制研究

Effect of thyrotropin on nitric oxide synthase in human microvascular endothelial cells and its mechanism

目的探讨在人微血管内皮细胞(HMEC-1)体外培养过程中,促甲状腺激素(TSH)对内皮型一氧化氮合酶(eNOS)表达的影响及其机制。方法分别以不同浓度TSH(0、10、100 mIU/ml)干预HMEC-1,应用qPCR法检测eNOS mRNA的表达水平,Western blotting法检测eNOS、磷酸化蛋白激酶B(p-AKT)、蛋白激酶B(AKT)、磷酸化细胞外信号调节激酶(P-ERK)、细胞外信号调节激酶(ERK)蛋白的水平。结果 (1)TSH可抑制HMEC-1中eNOS的表达,且呈剂量依赖性(P<0.05);(2)TSH可促进AKT、ERK的磷酸化(P<0.05)。结论促甲状腺激素可能通过激活AKT、ERK信号通路抑制人微血管内皮细胞一氧化氮合酶的表达。

Objective To investigate the effect of thyroid stimulating hormone （TSH） on the ex- pression of endothelial nitric oxide synthase （eNOS） and its mechanism in human microvascular endothelial cells （HMEC-1） in vitro culture. Methods Different concentrations of TSH （0, 10, 50 mlU/ml） were used to intervene HMEC-I. The expression of eNOS mRNA was detected with quantitative polymerase chain reaction （qPCR） method. The protein expressions of eNOS, phosphorylated protein kinase B （p-AKT） , protein kinase B （AKT） , phosphorylated extracellular signal-regulated kinase （P-ERK） , and extracellular signal-regulated kinase （ERK） were determined with Western Blot. Results （1） The expression level of eNOS was significantly decreased by TSH in dose-dependent manner （P 〈 0. 05 ）. （2） TSH could promote the phosphorylation of AKT and ERK （P 〈 0. 05）. Conclusions Thyroid-stimulating hormone may inhibit the expression of nitric oxide synthase in human microvascular endothelial cells by activating AKT and ERK signaling pathways.