甲状腺素及丙硫氧嘧啶对人脐静脉内皮细胞一氧化氮合酶表达的影响  

Effects of thyroxine and propylthiouracil on nitric oxide synthase expression in human umbilical vein endothelial cells

作　　者：贾海晗 景淼 刘畅 高云雁 刘岚春 刘鹏[1] Jia Haihan;Jing Miao;Liu Chang;Gao Yunyan;Liu Lanchun;Liu Peng(Institute of Iodine Deficiency Disorders,Center for Endemic Disease Control,Chinese Center for Disease Control and Prevention,Harbin Medical University,Harbin 150081,China;Heilongjiang Cancer Center,Cancer Hospital Affiliated to Harbin Medical University,Harbin 150081,China)

机构地区：[1]哈尔滨医科大学中国疾病预防控制中心地方病控制中心碘缺乏病防治研究所,150081 [2]哈尔滨医科大学附属肿瘤医院黑龙江省癌症中心,150081

出　　处：《中华地方病学杂志》2020年第3期168-171,共4页Chinese Journal of Endemiology

基　　金：国家自然科学基金(81773370);哈尔滨医科大学伍连德基金(WLD-1409)。

摘　　要：目的探讨甲状腺素(T4)及丙硫氧嘧啶(PTU)对人脐静脉内皮细胞(HUVEC细胞)一氧化氮合酶表达的影响。方法体外培养HUVEC细胞,实验分为6组,分别为对照组(未加T4及PTU),10^-9、10^-7、10^-4 mol/L T4组,PTU组(5μg/ml PTU),10^-4 mol/L T4+PTU组,作用时间为24 h。采用CCK-8法检测细胞活性;硝酸还原酶法检测细胞培养液一氧化氮(NO)、总一氧化氮合酶(TNOS)、内皮型一氧化氮合酶(eNOS)含量;蛋白质印迹法检测eNOS蛋白表达水平。结果6组细胞存活率[(100.00±0.00)%、(96.73±1.17)%、(86.20±7.54)%、(47.37±9.10)%、(53.37±5.47)%、(53.40±8.84)%]比较差异有统计学意义(F=29.42,P<0.05);与对照组比较,10^-7、10^-4 mol/L T4组,PTU组和10^-4 mol/L T4+PTU组细胞存活率显著降低(P均<0.05)。6组细胞培养液NO、TNOS含量比较差异有统计学意义(F=3.93、3.46,P均<0.05);与对照组比较,PTU组TNOS含量显著降低(P<0.05);与10^-4 mol/L T4组比较,PTU、10^-4 mol/L T4+PTU组NO含量显著降低(P均<0.05)。6组细胞培养液eNOS含量及蛋白表达水平比较差异无统计学意义(F=0.24、0.17,P均>0.05)。结论高浓度T4可造成体外培养HUVEC细胞活性的损伤,PTU通过调节NO、TNOS起到对其的缓解作用,具体作用机制还需进一步分子生物实验的深入研究。Objective To investigate the effects of thyroxine(T4)and propylthiouracil(PTU)on nitric oxide synthase expression in human umbilical vein endothelial cells(HUVEC cells).Methods HUVEC cells were cultured in vitro,the experiment was divided into 6 groups:control(without T4 and PTU),10^-9,10^-7,10^-4 mol/L T4,PTU(5μg/ml PTU),and 10^-4 mol/L T4+PTU,the action time was 24 h.CCK-8 method was used to detect cell activity;nitrate reductase method was used to detect nitric oxide(NO),total nitric oxide synthase(TNOS)and endothelial nitric oxide synthase(eNOS)contents;Western blotting was used to detect eNOS protein expression level.Results The differences of cell survival rates[(100.00±0.00)%,(96.73±1.17)%,(86.20±7.54)%,(47.37±9.10)%,(53.37±5.47)%,(53.40±8.84)%]among the 6 groups were statistically significant(F=29.42,P<0.05).Compared with the control group,the cell survival rates of 10^-7,10^-4 mol/L T4,PTU,and 10-4 mol/L T4+PTU groups were significantly reduced(P<0.05).There were statistically significant differences in the contents of NO and TNOS among the 6 groups(F=3.93,3.46,P<0.05).Compared with the control group,the TNOS content in PTU group was significantly reduced(P<0.05);compared with the 10^-4 mol/L T4 group,the NO contents in PTU and 10^-4 mol/L T4+PTU groups were significantly reduced(P<0.05).There were no significant differences in eNOS content and protein expression level among the 6 groups(F=0.24,0.17,P>0.05).Conclusions High concentration of T4 can cause damage to the activity of HUVEC cells cultured in vitro,and PTU can alleviate it by regulating NO and TNOS.The specific mechanism of action still needs to be further studied in molecular biological experiments.

关 键 词：甲状腺素 一氧化氮 丙硫氧嘧啶 人脐静脉内皮细胞 

分 类 号：R54[医药卫生—心血管疾病]