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目的探讨氢气在子前期胎盘缺血/再灌注细胞模型中的细胞保护作用及可能机制。方法体外培养的人绒毛膜癌细胞株JAR分为空白组、常氧培养条件下含饱和氢气组(常氧+H2组)、缺血/再灌注模型组(模型组)、缺血/再灌注饱和氢气干预组(模型+H2组)和缺血/再灌注维生素C阳性对照组(模型+VC组)。模型组、模型+H2组和模型+VC组。细胞在缺氧(5%CO2、95%N2)环境下无糖无血清培养2 h后,更换完全培养基,在常氧(5%CO2、95%空气)环境下培养,建立缺血/再灌注细胞模型,在缺氧及复氧期分别给予氢气或维生素C等干预。各组细胞在完全培养基中培养24 h后,采用MTT法检测细胞存活率,化学比色法检测细胞内丙二醛(MDA)含量及超氧化物歧化酶(SOD)的活性,Real-Time PCR检测SOD mRNA的表达;各组细胞在完全培养基培养1 h后,采用荧光探针2’,7’-二氢二氯荧光素二乙酸酯(DCFH-DA)检测细胞内活性氧类(ROS)水平。结果与模型组比较,模型+H2组细胞存活率升高(P<0.05),ROS产生及MDA含量明显减少(P<0.01,P<0.05),SOD活性增强(P<0.01),SOD mRNA表达水平升高(P<0.05)。结论氢气对子前期胎盘缺血/再灌注细胞模型有细胞保护作用,其机制可能是通过上调SOD-mRNA的转录、增强SOD酶活性及降低细胞内ROS水平,从而降低细胞氧化应激水平。
Objective To investigate the protective effect of hydrogen on placental ischemia / reperfusion cell model and its possible mechanism. Methods Human choriocarcinoma cell line JAR was divided into blank group, normoxia group (normoxia + H2 group), ischemia / reperfusion model group (model group), ischemia / reperfusion Saturated hydrogen intervention group (model + H2 group) and ischemia / reperfusion vitamin C positive control group (model + VC group). Model group, model + H2 group and model + VC group. Cells were cultured in sugar-free and serum-free medium under hypoxia (5% CO2, 95% N2) for 2 h. The complete medium was replaced and cultured under normoxia (5% CO2, 95% Perfused cell model, respectively, in hypoxia and reoxygenation were given hydrogen or vitamin C and other interventions. The cells in each group were cultured in complete medium for 24 h. The cell viability was detected by MTT assay. The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) PCR was used to detect the expression of SOD mRNA. After cultured in complete medium for 1 h, the cells were incubated with fluorescent probe 2 ’, 7’-dihydrofluorescein diacetate (DCFH-DA) for detection of intracellular reactive oxygen species (ROS) levels. Results Compared with model group, the cell viability in model group and H2 group increased (P <0.05), ROS production and MDA content decreased (P <0.01, P <0.05), SOD activity increased Elevated levels (P <0.05). Conclusions Hydrogen can protect cells against placental ischemia / reperfusion in early stage. Its mechanism may be through up-regulating the transcription of SOD-mRNA, enhancing the activity of SOD and decreasing the level of intracellular ROS, thereby reducing the level of cellular oxidative stress.