铁死亡参与无机砷致人肝星状细胞活化的研究  

The Effect of Ferroptosis on the Activation of Human Hepatic Stellate Cells Induced by Inorganic Arsenic

作　　者：迪丽娜尔·亚尔麦麦提 黄菲 丁关鑫 赵丽君 吴顺华 Dilinaer·Yaermaimaiti;HUANG Fei;DING Guanxin(Department of Public Health,Xinjiang Medical University,Xinjiang 830011,China)

机构地区：[1]新疆医科大学公共卫生学院流行病与卫生统计学教研室,乌鲁木齐830011 [2]新疆医科大学公共卫生学院劳动卫生与环境卫生学教研室

出　　处：《医学研究杂志》2024年第1期45-49,共5页Journal of Medical Research

基　　金：国家自然科学基金地区科学基金资助项目(82160650);新疆维吾尔自治区研究生科研创新项目(XJ2022G165)。

摘　　要：目的研究亚砷酸钠(NaAsO 2)对人肝星状细胞(LX-2)铁死亡的影响。方法体外培养LX-2细胞,采用成组设计将不同浓度NaAsO 2(5μmol/L、10μmol/L、15μmol/L)染毒LX-2细胞24h,构建体外NaAsO 2致LX-2活化模型,同时设立对照组;透射电子显微镜(transmission electron microscope,TEM)观察NaAsO 2处理LX-2细胞线粒体结构变化情况;荧光显微镜和荧光酶标仪检测细胞Fe 2+水平;比色法检测丙二醛(malondialdehyde,MDA)含量;蛋白免疫印迹法检测溶质载体家族7号成员11(solute carrier family number 7member 11,SLC7A11)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)、α-平滑肌肌动蛋白(α-SMA)蛋白表达水平。结果TEM结果显示,NaAsO 2组细胞线粒体膜完整性破损,线粒体嵴减少和消失;此外,与对照组比较,NaAsO 2处理组Fe 2+水平均升高(P<0.05);不同剂量NaAsO 2组MDA含量差异均有统计学意义(F=7.18,P均<0.05)。其中,与对照组比较,5、15μmol/L NaAsO 2组LX-2细胞MDA含量均高于对照组(P<0.05);在翻译水平上,纤维化指标α-SMA蛋白表达水平随着NaAsO 2剂量的升高而表达上调(P<0.05),铁死亡指标SLC7A11和GPX4蛋白表达水平随着NaAsO 2剂量的升高而减少(P<0.05)。结论铁死亡参与NaAsO 2致LX-2细胞活化过程。Objective To investigate the effect of NaAsO 2 on ferroptosis in human hepatic stellate cells(LX-2).Methods LX-2 cells were cultured in vitro,and different concentrations of NaAsO 2(5μmol/L,10μmol/L,15μmol/L)were infected with LX-2 cells for 24h in a group design to construct the activation model of LX-2 induced by NaAsO 2 in vitro,and a control group was set up.Mitochondrial structure of LX-2 cells treated with NaAsO 2 was observed by transmission electron microscopy(TEM).Fe 2+levels were detected by fluorescence microscope and fluorescent enzyme label.The content of malondialdehyde(MDA)was determined by the colorimetric method.The protein expression levels of SLC7A11,GPX4,andα-smooth muscle actin(α-SMA)were detected by Western blot.Results TEM showed that mitochondrial membrane integrity was damaged and mitochondrial ridges were reduced and disappeared in the NaAsO 2group.In addition,compared with the control group,Fe 2+levels in NaAsO 2 treatment groups were increased(P<0.05).There were statistically significant differences in MDA content among different doses of NaAsO 2groups(F=7.18,P<0.05).Compared with the control group,MDA content of LX-2 cells in 5 and 15μmol/L NaAsO 2groups was higher than that in the control group(P<0.05).At the translation level,the expression of fibrosis indexα-SMA protein level was up-regulated with the increase of NaAsO 2dose(P<0.05),the protein expression levels of ferroptosis index SLC7A11 and GPX4decreased in a dose-dependent manner with the increase of the dose of NaAsO 2(P<0.05).Conclusion Ferroptosis is involved in the activation of LX-2 cells induced by NaAsO 2.

关 键 词：亚砷酸钠 人肝星状细胞 铁死亡 

分 类 号：R12[医药卫生—环境卫生学]