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目的探讨全氟辛烷磺酰基化合物(PFOS)致人胚胎肝(L-02)细胞凋亡的作用及其机制。方法将处于对数生长期的L-02细胞分别暴露于终浓度为0(对照)、50、100、150、200μmol/L的PFOS溶液培养24 h。采用MTT法检测细胞活性,采用流式细胞术检测细胞凋亡率,并测定细胞线粒体内活性氧(ROS)、谷胱甘肽(GSH)、丙二醛(MDA)和超氧化物歧化酶(SOD)的水平及线粒体膜电位,采用q RT-PCR法检测凋亡相关基因caspase-3、caspase-9、bax、bcl-2 m RNA的表达水平。结果与对照组比较,150、200μmol/L PFOS染毒L-02细胞的存活率均降低,而100、150、200μmol/L PFOS染毒组L-02细胞的凋亡率均增加,差异均有统计学意义(P<0.05);且随着PFOS染毒浓度的升高,L-02细胞的存活率呈下降趋势,而凋亡率呈上升趋势。与对照组比较,各浓度PFOS染毒组L-02细胞内ROS和MDA水平均增加,而各浓度PFOS染毒组L-02细胞内SOD水平和100、150、200μmol/L PFOS染毒组L-02细胞内GSH水平和线粒体膜电位均较低,差异均有统计学意义(P<0.05);且随着PFOS染毒浓度的升高,L-02细胞内ROS和MDA水平均呈上升趋势,而SOD、GSH水平和线粒体膜电位均呈下降趋势。与对照组比较,各浓度PFOS染毒组L-02细胞中caspase-3、caspase-9、Bax m RNA的表达水平均较高,而bcl-2、bcl-2/bax值均较低,除50μmol/L PFOS染毒组caspase-9外,差异均有统计学意义(P<0.05﹚。结论在本实验剂量下,PFOS暴露可引起L-02细胞凋亡,其机制与PFOS诱导的氧化损伤和线粒体凋亡途径有关。
Objective To investigate the effect and mechanism of perfluorooctane sulfonate (PFOS) on human embryonic liver (L-02) cell apoptosis. Methods L-02 cells in logarithmic growth phase were exposed to a final concentration of 0 (control), 50, 100, 150, 200μmol / L of PFOS solution for 24 h. The cell viability was measured by MTT assay. The apoptosis rate was detected by flow cytometry. The levels of reactive oxygen species (ROS), glutathione (GSH), malondialdehyde (MDA) and superoxide dismutase SOD, and mitochondrial membrane potential. The expression of caspase-3, caspase-9, bax and bcl-2 mRNA were detected by q RT-PCR. Results Compared with the control group, the survival rate of L-02 cells treated with 150,200 μmol / L PFOS was decreased, while the apoptosis rates of L-02 cells were increased with 100, 150 and 200 μmol / L PFOS exposure (P <0.05). With the increase of PFOS concentration, the survival rate of L-02 cells decreased and the apoptosis rate increased. Compared with the control group, the levels of ROS and MDA in L-02 cells in PFOS-treated groups increased at all concentrations, while the levels of SOD in L-02 cells in PFOS-treated groups were significantly higher than those in PFOS-exposed groups (P <0.05). The levels of ROS and MDA in L-02 cells were increased with the increase of PFOS exposure, and the levels of GSH and mitochondrial membrane potential in L-02 cells were lower , While SOD, GSH levels and mitochondrial membrane potential showed a downward trend. Compared with the control group, the expression levels of caspase-3, caspase-9 and Bax m RNA in L-02 cells at various concentrations of PFOS were lower than those in the control group, while the bcl-2 and bcl-2 / 50μmol / L PFOS exposure group caspase-9, the differences were statistically significant (P <0.05). Conclusions PFOS exposure can induce L-02 cell apoptosis at the dose of this experiment. The mechanism is related to the oxidative damage induced by PFOS and the mitochondrial apoptosis pathway.