【摘要】 目的 观察亚砷酸钠（NaAsO₂）对人肝星状细胞的激活作用，并探讨其与铁死亡的关系。方法 体外培养人肝星状细胞LX-2，将细胞分为NaAsO₂组和对照组，NaAsO₂组分别加入5、10、15μmol/LNaAsO₂培养24 h，对照组正常培养。倒置显微镜下观察细胞形态变化；比色法检测谷胱甘肽（GSH）含量；荧光探针法检测细胞脂质活性氧（ROS）;RT-PCR法检测铁死亡指标溶质载体家族7成员11（SLC7A11）、谷胱甘肽过氧化物酶4（GPX4）以及纤维化指标α平滑肌肌动蛋白（α-SMA）和Ⅰ型胶原蛋白（CollagenⅠ） mRNA；采用Spearman相关分析法分析纤维化指标与铁死亡指标的相关性。结果 NaAsO₂组中随着NaAsO₂浓度升高，细胞触角逐渐回缩，细胞数量减少，细胞间距变宽。与对照组比较，NaAsO₂不同浓度组细胞GSH含量均减少，其中15μmol/LNaAsO₂组GSH含量低于10、5μmol/LNaAsO₂组，10μmol/LNaAsO₂组低于5μmol/LNaAsO₂组（P均<0.05）。与对照组比较，NaAsO₂不同浓度组脂质ROS荧光强度增强。与对照组比较，5、10、15μmol/LNaAsO₂组α-SMA、CollagenⅠmRNA表达水平均升高，且15μmol/LNaAsO₂组高于5、10μmol/LNaAsO₂组，10μmol/LNaAsO₂组高于5μmol/LNaAsO₂组（P均<0.05）；与对照组比较，5、10、15μmol/LNaAsO₂组SLC7A11、GPX4 mRNA表达水平均降低，且15μmol/LNaAsO₂组低于10、5μmol/LNaAsO₂组，10μmol/LNaAsO₂组低于5μmol/LNaAsO₂组。α-SMA mRNA与GPX4、SLC7A11 mRNA表达呈负相关（r分别为-0.72、-0.92,P均<0.01）,CollagenⅠmRNA与GPX4、SLC7A11 mRNA表达亦呈负相关（r分别为-0.67、-0.90,P均<0.01）。结论 NaAsO₂能够改变肝星状细胞形状，促进细胞纤维化基因表达，引起肝星状细胞激活；在此过程中，SLC7A11、GPX4基因表达下调导致ROS堆积，促进肝星状细胞铁死亡；上述作用随着NaAsO₂浓度升高而增强。更多还原

【Abstract】 Objective To observe the effect of sodium arsenite（NaAsO₂） on the activation of human hepatic stellate cells and its relationship with ferroptosis. Methods Human liver stellate cells LX-2 were cultured in vitro, and were di-vided in the NaAsO₂ group and control group. Cells in the NaAsO₂ group were treated with 5, 10 and 15 μmol/LNaAsO₂ for 24 h, respectively, and cells in the control group underwent normal culture. The morphological changes of cells were observed under inverted microscope. The content of glutathione（GSH） was determined by colorimetry. Fluorescent probe was used to detect cell lipid reactive oxygen species（ROS）. Ferroptosis markers SLC7A11 and glutathione peroxidase 4（GPX4）, fibrosis markers α smooth muscle actin（α-SMA） and CollagenⅠmRNA were detected by RT-PCR. Spearman correlation analysis was used to analyze the correlation between fibrosis index and ferroptosis index. Results With the in-crease of NaAsO₂ concentration, the cell antennae gradually retreated, the number of cells decreased, and the intercellular space expanded. Compared with the control group, the GSH content of cells in different NaAsO₂ concentration groups de-creased, and the GSH content in the 15μmol/LNaAsO₂ group was lower than that in the 10 and 5 μmol/LNaAsO₂ groups, and that in the 10 μmol/LNaAsO₂ group was lower than that in the 5 μmol/LNaAsO₂ group（all P<0. 05）. Compared with the control group, the fluorescence intensity of lipid ROS in the NaAsO₂ groups was enhanced. Compared with the control group, α-SMA and Collagen1 mRNA expression levels in the 5, 10 and 15 μmol/LNaAsO₂ groups increased, and those in the 15 μmol/LNaAsO₂ group were higher than those in the 10 and 5 μmol/LNaAsO₂ groups; those in the 10 μmol/LNa-AsO₂ group were higher than those in the 5 μmol/LNaAsO₂ group（all P<0. 05）. Compared with the control group, the mRNA expression levels of SLC7A11 and GPX4 in the 5, 10 and 15 μmol/LNaAsO₂ groups decreased, and those in the 15 μmol/LNaAsO₂ group were lower than those in the 10 and 5 μmol/LNaAsO₂ groups; those in the 10 μmol/LNaAsO₂ group were lower than those in the 5 μmol/LNaAsO₂ group. The α-SMA mRNA was negatively correlated with GPX4 and SLC7A11 mRNA expression（r=-0. 72,-0. 92, respectively; both P<0. 01）, and CollagenⅠ mRNA was negatively corre-lated with GPX4 and SLC7A11 mRNA expression（r=-0. 67,-0. 90, respectively; both P<0. 01）. Conclusions With the increase of NaAsO₂ concentration, the expression level of fibrosis gene increases gradually, which causes the activation of hepatic stellate cells. In this process, the down-regulated expression of SLC7A11 and GPX4 genes causes ROS accumu-lation and promotes ferroptosis in hepatic stellate cells.更多还原