硒化合物对人脐带间充质干细胞ROS和NO生成及NOS活性的影响

Effect of selenium compounds on ROS and NO production and NOS activity in human umbilical cord mesenchymal stem cells

目的:通过观察不同的硒化合物对人脐带间充质干细胞(MSCs)一氧化氮(NO)、活性氧(ROS)的生成及一氧化氮合酶(NOS)活性的影响,初步探讨硒的免疫调节作用机制。方法:在MSCs培养的过程中,分别添加0.1μmol/L和0.5μmol/L亚硒酸钠[Se(Ⅳ)]、硒代胱氨酸(Se-Cys)、纳米硒(Nano-Se),采用硝酸还原酶法检测24 h和48 h MSCs NO和NOS活性的变化,通过DCFH-DA荧光探针检测细胞内ROS。结果:在24 h,对照组、0.1μmol/L和0.5μmol/L Se(Ⅳ)处理MSCs的NO水平分别是(18.13±6.80)μmol/L、(20.93±5.68)μmol/L和(16.73±5.03)μmol/L。在48 h,0.1μmol/L和0.5μmol/L Se(IV)处理MSCs的NO水平分别是(17.20±9.11)μmol/L(P<0.05)和(9.98±4.35)μmol/L(P<0.01),明显低于对照组的(26.23±4.35)μmol/L。Se(IV)对ROS和NOS有明显的抑制作用,Se-Cys和Nano-Se对NO、ROS的生成和NOS活性没有明显影响。结论:Se(Ⅳ)对MSCs NO、ROS的生成和NOS活性有明显的抑制效应。

AIM：To investigate the effects of selenium（Se） compounds on the generation of nitric oxide（NO） and reactive oxygen species（ROS）,and the activity of nitric oxide synthase（NOS） in umbilical cord mesenchymal stem cells（MSCs）.METHODS： MSCs were cultured in the medium of DMEM/F12 containing 10% FBS and exposed to the compounds of selenium [Na2SeO3,Se（IV）],selenocysteine（Se-Cys） or nano-selenium（Nano-Se） at concentrations of 0.1 μmol/L to 0.5 μmol/L.The concentration of NO and the activity of NOS in treated MSCs were detected by the method of nitrate reductase assay.The fluorescent intensity of ROS was determined by DCFH-DA probe.RESULTS： The production of NO was（18.13±6.80） μmol/L in the control MSCs,（20.93±5.68） μmol/L in the MSCs treated with Se（Ⅳ） at concentration of 0.1 μmol/L and（16.73±5.03） μmol/L in the MSCs treated with Se（Ⅳ） at concentration of 0.5 μmol/L for 24 h.The production of NO was（17.20±9.11） μmol/L（P0.05） in the MSCs treated with Se（IV） at concentration of 0.1 μmol/L and（9.98±4.35） μmol/L（P0.01） in the MSCs treated with Se（IV） at concentration of 0.5 μmol/L for 48 h,which was significantly lower than that in the control MSCs [（26.23±4.35） μmol/L].No inhibitory effect of Nano-Se and Se-Cys on the production of NO in MSCs was observed.Compared with the control MSCs,Se（Ⅳ） at concentrations of 0.1 μmol/L and 0.5 μmol/L significantly inhibited the generation of ROS and the activity of NOS in MSCs at 24 h and 48 h.CONCLUSION： Se（Ⅳ） inhibits NO/ROS production and NOS activity in a dose-dependent manner in MSCs.