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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。
OBJECTIVE: To investigate the effects of different selenium compounds on the production of nitric oxide (NO), reactive oxygen species (ROS) and the activity of nitric oxide synthase (NOS) in human umbilical cord mesenchymal stem cells (MSCs) Regulate the mechanism of action. Methods: Sepharose [Se (Ⅳ)], Se-Cys and Nano-Se were added into the culture medium of MSCs at the concentrations of 0.1μmol / L and 0.5μmol / Nitric acid reductase was used to detect the changes of NO and NOS activities in MSCs at 24 h and 48 h. The intracellular ROS was detected by DCFH-DA fluorescent probe. Results: The levels of NO in MSCs treated with 0.1 μmol / L and 0.5 μmol / L Se (Ⅳ) for 24 h were (18.13 ± 6.80) μmol / L and (20.93 ± 5.68) μmol / L and 5.03) μmol / L. The NO levels of MSCs treated with 0.1 μmol / L and 0.5 μmol / L Se (IV) for 48 h were (17.20 ± 9.11) μmol / L (P <0.05) and (9.98 ± 4.35) μmol / L ), Which was significantly lower than that of the control group (26.23 ± 4.35) μmol / L. Se (IV) significantly inhibited ROS and NOS. Se-Cys and Nano-Se had no significant effect on NO and ROS production and NOS activity. Conclusion: Se (Ⅳ) significantly inhibits the production of NO and ROS and the activity of NOS in MSCs.