摘要
目的 探讨D-半乳糖(D-gal)诱导的心肌细胞衰老是否存在铁死亡及铁死亡抑制剂Ferrostatin-1(Fer-1)能否延缓心肌细胞衰老。方法 通过不同水平(0、5、10、20、40、80、100 g/L)D-gal诱导H9C2心肌细胞损伤从而制备心脏衰老模型,采用MTT法检测细胞活力,确定后续实验采用的D-gal质量浓度。将细胞分为Control组、D-gal组和Fer-1组。MTT法检测细胞活力;DCFH-DA法检测细胞内活性氧(ROS)水平;微板法检测细胞内还原型谷胱甘肽(GSH)含量;硫代巴比妥酸法检测细胞内丙二醛(MDA)含量;Western blot检测溶质载体家族7成员11(SLC7A11)、谷胱甘肽过氧化物酶4(GPx4)、P53蛋白表达水平;微量法检测细胞内β-半乳糖苷酶(β-GAL)活性。结果 MTT法结果显示,细胞活力随D-gal质量浓度升高而降低(P<0.05),后续实验采用D-gal质量浓度为20 g/L。与D-gal组相比,Fer-1组细胞活力增加(P<0.05);与Control组相比,D-gal组中ROS、MDA含量、P53蛋白表达水平、β-GAL活性增强,GSH含量、SLC7A11、GPx4蛋白表达水平降低(P<0.05);与D-gal组相比,Fer-1组中ROS、MDA含量、P53蛋白表达水平、β-GAL活性降低,GSH、SLC7A11、GPx4蛋白表达升高(P<0.05)。结论 D-gal诱导的心肌细胞衰老过程中发生了铁死亡,Ferrostatin-1可抑制铁死亡,从而延缓心肌细胞衰老。
Objective To investigate the existence of ferroptosis in cardiomyocyte senescence induced by D-galactose(D-gal) and whether ferroptosis inhibitor ferrostatin-1(Fer-1) can delay cardiomyocyte senescence.Methods Cardiomyocyte injury was induced by D-gal(0,5,10,20,40,80 and 100 g/L) to simulate cardiac aging model.After cells were treated with different concentrations of D-gal for 24 hours,the cell viability was detected by MTT method to determine the concentration of D-gal in the follow-up experiment.The cells were divided into the control group,the D-gal group and the Fer-1 group.The cell viability was detected by MTT method.The level of intracellular reactive oxygen species(ROS) was detected by DCFH-DA method.The intracellular glutathione(GSH) content was detected by microplate method.Intracellular malondialdehyde(MDA) content was detected by thiobarbituric acid method.The protein expression levels of SLC7A11,GPx4 and P53 were detected by Western blot assay.The activity of intracellular β-galactosidase(β-GAL) was detected by microassay.Results MTT assay showed that the cell viability decreased with the increase of D-gal concentration(P <0.05).The concentration of D-gal was 20 g/L in subsequent experiments.Compared with the D-gal group,the cell viability was increased in the Fer-1 group(P <0.05).Compared with the control group,ROS and MDA contents,P53protein expression level and β-GAL activity were increased in the D-gal group,while GSH content,SLC7A11 and GPx4 protein expression levels were decreased(P<0.05).Compared with the D-gal group,ROS and MDA contents,P53 protein expression level and β-GAL activity were decreased in the Fer-1 group,while GSH,SLC7A11 and GPx4 protein expressions were increased(P<0.05).Conclusion Ferroptosis occurs during D-gal-induced cell senescence,and Fer-1 can inhibit iron death and delay myocardial senescence.
作者
熊喜成
王一平
王刚
张甜
包雅丽
迪娜·艾尼瓦尔
孙湛
XIONG Xicheng;WANG Yiping;WANG Gang;ZHANG Tian;BAO Yali;Di na·AINIWAER;SUN Zhan(Department of Pathophysiology,College of Basic Medicine,Xinjiang Medical University,Urumqi 830000,China;Xinjiang Key Laboratory of Molecular Biology for Endemic Diseases)
出处
《天津医药》
CAS
北大核心
2023年第1期19-23,共5页
Tianjin Medical Journal
基金
新疆维吾尔自治区自然科学基金资助项目(2019D01C207)
新疆地方与民族高发病教育部重点实验室开放基金(KF2021-5)。
