摘要
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: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·kg^(-1));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·kg^(-1));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1β(IL-1β)和肿瘤坏死因子α(TNF-α)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: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(P<0.01);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(P<0.01)。与对照组比较,LPS组小鼠血清中CRE和BUN水平明显升高(P<0.01);与LPS组比较,LPS+罗格列酮组小鼠CRE和BUN水平明显降低(P<0.01)。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(P<0.01),GPX4和SLC7A11 mRNA和蛋白表达水平降低(P<0.01),IL-6、IL-1β和TNF-αmRNA表达水平升高(P<0.01);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(P<0.01),GPX4和SLC7A11 mRNA及蛋白水平明显升高(P<0.01),IL-6、IL-1β和TNF-αmRNA表达水平降低(P<0.01)。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。
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·kg^(-1) LPS intraperitoneally;the mice in LPS+rosiglitazone group were injected with 0.5 mg·kg^(-1) 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-1β(IL-1β),and tumor necrosis factor-α(TNF-α)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(P<0.01);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(P<0.01).Compared with control group,the levels of CRE and BUN in serum of the mice in LPS group were increased(P<0
作者
杨巧玲
符璐
石雨
叶严珏
卢日峰
刘永
尹俐
YANG Qiaoling;FU Lu;SHI Yu;YE Yanjue;LU Rifeng;LIU Yong;YIN Li(Department of Biopharmaceutical Sciences,School of Pharmacy and Bioengineering,Chongqing University of Technology,Chongqing 400054,China;Department of Nephrology,Xinqiao Hospital,Army Military Medical University,Chongqing 400037,China;Department of Hygiene Toxicology,School of Public Health,Jilin University,Changchun 130021,China)
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2023年第2期351-359,共9页
Journal of Jilin University：Medicine Edition
基金
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)。
关键词
急性肾损伤
长链脂酰CoA合成酶4
铁死亡
罗格列酮
Acute kidney injury
Acyl CoA synthetase long-chain family member 4
Ferroptosis
Rosiglitazone
