铁死亡在百草枯致肺泡上皮细胞间充质化中的作用研究  

Studies on the role of iron death in paraquat-induced mesenchymalization of alveolar epithelial cells

作　　者：苏艺伟 方文馨 唐侍豪 李广珍 王致[1] Su Yiwei;Fang Wenxin;Tang Shihao;Li Guangzhen;Wang Zhi(Key Laboratory of Occupational Environment and Health,Guangzhou Twelfth People's Hospital,Guangzhou 510620,China;School of Basic Medicine and Public Health,Jinan University,Guangzhou 510632,China)

机构地区：[1]广州市第十二人民医院职业环境与健康重点实验室,广州510620 [2]暨南大学基础医学与公共卫生学院,广州510632

出　　处：《中华劳动卫生职业病杂志》2025年第2期110-116,共7页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：广州市科学技术局市校(院)企联合资助项目(2023A03J0977);广东省基础与应用基础研究基金联合基金项目(2021A1515110663);广州市科学技术局重点研发计划项目(202206010061);广州市科学技术局市校(院)企联合资助项目(2023A03J0502)。

摘　　要：目的探讨铁死亡在百草枯(PQ)致肺泡上皮细胞间充质化(EMT)中的作用。方法于2023年8月,采用CCK8法筛选出合适的PQ及脂血抑素-1(Lip-1)处理浓度,将RLE-6TN细胞分为对照组、PQ组和铁死亡抑制组,PQ组给予200μmol/L PQ溶液,铁死亡抑制组给予200μmol/L PQ溶液和0.1μmol/L Lip-1溶液,对照组加入等量细胞培养基。于染毒后12、24、48 h观察各组细胞形态改变及迁移活力,检测各组细胞亚铁离子(Fe^(2+))、活性氧自由基(ROS)、谷胱甘肽(GSH)、超氧化物歧化酶(SOD)、丙二醛(MDA)含量;同时采用实时荧光定量聚合酶链反应(qRT-PCR)和蛋白免疫印迹(Western blot)法测定各组细胞E-钙黏蛋白(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)、胶原蛋白I(Collagen I)分子的表达情况。多组间差异比较采用方差分析,进一步两两比较用Bonferroni法。结果用CCK8检测细胞活力,结果显示200μmol/L PQ+0.1μmol/L Lip-1溶液处理RLE-6TN细胞后,细胞存活率为56.6%。铁死亡抑制组RLE-6TN细胞在染毒24和48 h后迁移活力较PQ组弱,且细胞EMT改变程度较PQ组减轻。铁死亡抑制组细胞在染毒12、24和48 h后,各时间点Fe^(2+)浓度、ROS荧光强度值、MDA含量均较PQ组各相应时间点均降低(P<0.05/3),而其GSH和SOD浓度均较PQ组各相应时间点均升高(P<0.05/3)。结果显示,与对照组比较,PQ组和铁死亡抑制组细胞E-cadherin mRNA和蛋白表达量均降低(P<0.05/3),与PQ组比较,铁死亡抑制组细胞E-cadherin mRNA和蛋白表达量升高(P<0.05/3);与对照组比较,PQ组和铁死亡抑制组细胞α-SMA、CollagenⅠmRNA和蛋白表达量均升高(P<0.05/3),与PQ组比较,铁死亡抑制组细胞α-SMA、CollagenⅠmRNA和蛋白的表达量降低(P<0.05/3)。结论铁死亡参与PQ致肺泡上皮细胞EMT过程,抑制铁死亡,降低细胞氧化损伤,减轻细胞EMT程度。ObjectiveTo investigate the role of iron death in paraquat(PQ)-induced alveolar epithelial mesangialization(EMT).MethodsIn August 2023,the appropriate PQ staining concentration as well as the intervention concentration of lipoinhibitor-1(Lip-1)were screened by CCK8 method.The RLE-6TN cells were divided into three groups,which were control group,PQ group and iron death inhibition group,200μmol/L PQ solution was given to the PQ group,and PQ 200μmol/L and 0.1μmol/L Lip-1 solution was given to the iron death inhibition group,the control group was added the same amount of cell medium.morphological changes and migratory viability of the cells in each group were observed at 12 h,24 h and 48 h after the poisoning,and the contents of ferrous ions(Fe^(2+)),reactive oxygen radicals(ROS),glutathione(GSH),superoxide dismutase(SOD),and malondialdehyde(MDA)were detected in each group;meanwhile,qRT-PCR and western-blot were used to determine the molecular expression of E-cadherin,α-smooth muscle actin(α-SMA),and Collagen I in the cells in each group.The difference between group was compared by ANOVA,and the further pairwise comparison was conducted by Bonferroni method.ResultsCell viability was detected using CCK8,and the results showed that the cell survival rate of RLE-6TN cells treated with 200μmol/L PQ+0.1μmol/L Lip-1 solution was 56.6%.The migration activity of RLE-6TN cells in the iron death inhibition group was weaker than that in the PQ group after 24 and 48 hours of exposure,and the degree of EMT changes in the cells was reduced compared to the PQ group.After 12,24,and 48 hours of exposure,the Fe^(2+)concentration,ROS fluorescence intensity,and MDA content in the iron death inhibition group decreased compared to the corresponding time points in the PQ group(P<0.05/3),while compared with PQ group,the GSH concentration and SOD concentration increased compared to the corresponding time points in the PQ group(P<0.05/3).The results showed that compared with the control group,the expression levels of E-cadherin mRNA an

关 键 词：百草枯 铁死亡 上皮间充质化 肺泡上皮细胞 脂血抑素-1 

分 类 号：R73[医药卫生—肿瘤]