【摘要】 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。更多还原

【Abstract】 Objective To understand the effect of oxidative stress on the apoptosis of L-02 cells induced by PFOS. Methods L-02 cells were exposed to 0,50,100,150 and 200 μmol/L of PFOS for 24 h. MTT method was used to measure the survival of L-02 cells,the cell apoptosis was measured by flow cytometry,the L-02 cells homogenates were prepared to detect the levels of reactive oxygen species(ROS),malondialdehyde(MDA),superoxide dismutase(SOD) and glutathione hormone(GSH) in spectro-photometric assay. Rhodamine 123 fluorescent probe was used to detect the mitochondrial membrane potential altering in L-02 cells. The expression levels of caspase-3,caspase-9,bcl-2 and bax m RNA were examined by q RT-PCR. Results Compared with the control group,the livability of L-02 cells decreased significantly in 150,200 μmol/L PFOS-treated groups(P<0.05﹚,the percentage of apoptosis increased significantly in 100,150 and 200 μmol/L PFOS-treated groups(P<0.05﹚,the livability tendency of L-02 cells decreased as the dose of PFOS increased,and the apoptosis rates in L-02 cells showed an increasing tendency. Compared with the control group,the levels of ROS and MDA increased significantly in PFOS-exposed groups(P<0.05),the levels of SOD decreased significantly in PFOS-exposed groups,the levels of GSH and MMP decreased significantly in100,150 and 200 μmol/L PFOS-treated groups(P<0.05),as the concentrations of PFOS increased and the levels of ROS and MDA showed an increased trend and the levels of SOD,GSH showed a declining trend. Compared with the control groups,except the level of caspase-9 in 50 μmol/L PFOS-treated groups,the levels of caspase-3,caspase-9,bax m RNA increased significantly in PFOS-exposed groups(P <0.05),the levels of bcl-2 m RNA and the bcl m RNA/bax m RNA value in PFOS-exposed groups were significantly lower(P<0.05﹚. Conclusion At the dose of PFOS exposed in the present study,PFOS can induce the L-02 cell apoptosis,and mitochondria mediated signaling pathway may be involved in it.更多还原