罗格列酮对脂多糖诱导小鼠急...胞铁死亡的抑制作用及其机制_杨巧玲.pdf

第 49 卷 第 2 期2023年 3 月吉林大学学报（医学版）Journal of Jilin University（Medicine Edition）Vol.49 No.2Mar.2023DOI：10.13481/j.1671587X.20230211罗格列酮对脂多糖诱导小鼠急性肾损伤肾小管上皮细胞铁死亡的抑制作用及其机制杨巧玲1,2,符璐1,石雨1,叶严珏1,卢日峰3,刘永2,尹俐1（1.重庆理工大学药学与生物工程学院生物制药系，重庆 400054；2.陆军军医大学新桥医院肾内科，重庆 400037；3.吉林大学公共卫生学院卫生毒理学教研室，吉林 长春 130021）摘要 目的目的：探讨罗格列酮对脂多糖（LPS）诱导的急性肾损伤（AKI）小鼠肾小管上皮细胞铁死亡的抑制作用，并阐明其作用机制。方法方法：18 只 C57BL/6 雄性小鼠随机分为对照组、LPS 组和LPS+罗格列酮组，每组 6 只。LPS 组和 LPS+罗格列酮组小鼠均腹腔注射 LPS（10 mg kg1）；LPS+罗格列酮组小鼠在 LPS注射前 30 min 尾静脉注射罗格列酮（0.5 mg kg1）；对照组小鼠注射与 LPS 组小鼠同体积的生理盐水。LPS 注射 24 h后处死小鼠，收集肾组织和血清。HE 染色观察各组小鼠肾组织病理形态表现，分光光度法检测各组小鼠血清中肌酐（CRE）和血尿素氮（BUN）水平，实时荧光定量 PCR（RT-qPCR）法检测各组小鼠肾组织中长链脂酰 CoA 合成酶 4（ACSL4）、谷胱甘肽过氧化物酶 4（GPX4）、溶质载体家族 7成员 11（SLC7A11）、白细胞介素 6（IL-6）、白细胞介素1（IL-1）和肿瘤坏死因子 （TNF-）mRNA 表达水平，Western blotting法检测各组小鼠肾组织中GPX4、SLC7A11和 ACSL4蛋白表达水平。结果结果：HE染色，与对照组比较，LPS组小鼠肾小管管腔内出现空泡结构，肾小管损伤评分明显升高（P0.01）；与 LPS 组比较，LPS+罗格列酮组小鼠肾组织中细胞排列紧密，管腔内基本无空泡，肾小管损伤评分明显降低（P0.01）。与对照组比较，LPS 组小鼠血清中 CRE 和 BUN 水平明显升高（P0.01）；与 LPS 组比较，LPS+罗格列酮组小鼠CRE 和 BUN 水平明显降低（P0.01）。RT-qPCR 法和 Western blotting 法，与对照组比较，LPS 组小鼠肾组织中 ACSL4 mRNA 和蛋白表达水平升高（P0.01），GPX4 和 SLC7A11 mRNA 和蛋白表达水平降低（P0.01），IL-6、IL-1 和 TNF-mRNA 表达水平升高（P0.01）；与 LPS 组比较，LPS+罗格列酮组小鼠肾组织中 ACSL4 mRNA 和蛋白表达水平明显降低（P0.01），GPX4 和SLC7A11 mRNA及蛋白水平明显升高（P0.01），IL-6、IL-1 和 TNF-mRNA 表达水平降低（P0.01）。结论结论：罗格列酮可通过调控 ACSL4改善 LPS诱导的 AKI小鼠肾小管上皮细胞铁死亡。关键词 急性肾损伤；长链脂酰 CoA合成酶 4；铁死亡；罗格列酮中图分类号 R-332;R285.5文献标志码 AInhibitory effect of rosiglitazone on ferroptosis of renal tubular epithelial cells in mice with acute renal injury induced by lipopolysaccharide and its mechanismYANG Qiaoling1,2,FU Lu1,SHI Yu1,YE Yanjue1,LU Rifeng3,LIU Yong2,YIN Li1（1.Department of Biopharmaceutical Sciences，School of Pharmacy and Bioengineering，Chongqing 文章编号 1671587X(2023)02035109收稿日期 20220509基金项目 重 庆 市 科 委 自 然 科 学 基 金 面 上 项 目（cstc2019jcyj-msxmX0427）；吉 林 省 科 技 厅 重 点 研 发 项 目（20200403029SF）；重庆市教委科学技术研究项目（KJQN201901139）；重庆理工大学科研启动基金项目（2019ZD35）作者简介 杨巧玲（1996），女，重庆市人，在读硕士研究生，主要从事肾脏疾病基础方面的研究。通信作者 尹 俐，副教授，硕士研究生导师（E-mail：）351第 49 卷 第 2 期 2023 年 3 月吉林大学学报（医学版）University of Technology，Chongqing 400054，China；2.Department of Nephrology，Xinqiao Hospital，Army Military Medical University，Chongqing 400037，China；3.Department of Hygiene Toxicology，School of Public Health，Jilin University，Changchun 130021，China）ABSTRACT Objective：To investigate the inhibitory effect of rosiglitazone on ferroptosis of renal tubular epithelial cells in the mice with acute kidney injury（AKI）induced by lipopolysaccharide（LPS），and to clarify its mechanism.Methods：A total of 18 C57BL/6 male mice were randomly divided into control group（n=6），LPS group（n=6），and LPS+rosiglitazone group（n=6）.The mice in LPS group and LPS+rosiglitazone group were injected with 10 mg kg1 LPS intraperitoneally；the mice in LPS+rosiglitazone group were injected with 0.5 mg kg1 rosiglitazone via the tail vein 30 min before LPS injection；the mice in control group were injected with the same volume of normal saline.The mice were sacrificed 24 h after LPS injection，and the kidney tissue and serum were collected.HE staining was used to observe the pathomorphology of kidney tissue of the mice in various groups；the levels of serum creatinine（CRE）and urea nitrogen（BUN）of the mice were detected by spectrophotometry；real-time fluorescence quantitative PCR（RT-qPCR）method was used to detect the expression levels of acyl CoA synthetase long-chain family member 4（ACSL4），glutathione peroxidase 4（GPX4），solute carrier family 7 member 11（SLC7A11），interleukin-6（IL-6），interleukin-1（IL-1），and tumor necrosis factor-（TNF-）mRNA in kidney tissue of the mice in various groups；Western blotting method was used to detect the expression levels of ACSL4，GPX4，and SLC7A11 proteins in kidney tissue of the mice in various groups.Results：The HE staining results showed that compared with control group，there were vacuoles in the renal tubule lumen of the mice in LPS group，the score of renal tubular injury was increased significantly（P0.01）；compared with LPS group，the renal tubular epithelial cells in kidney tissue of the mice in LPS+rosiglitazone group were closely arranged，there was almost no protein tube，and the score of renal tubular injury of the mice was decreased significantly（P0.01）.Compared with control group，the levels of CRE and BUN in serum of the mice in LPS group were increased（P0.01）；compared with LPS group，the levels of CRE and BUN in serum of the mice in LPS+rosiglitazone group were decreased significantly（P0.01）.The RT-qPCR and Western blotting results showed that compared with control group，the expression levels of ACSL4 mRNA and protein in kidney tissue of the mice in LPS group were increased（P0.01），the expression levels of GPX4 and SLC7A11 mRNA and proteins were decreased（P0.01），and the expression levels of IL-6，IL-1，and TNF-mRNA were increased（P0.01）；compared with LPS group，the expression levels of ACSL4 mRNA and protein in kidney tissue of the mice in LPS+rosiglitazone group were decreased（P0.01），the expression levels of GPX4 and SLC7A11 mRNA and proteins were increased