摘要
目的:探讨铁死亡对高糖损伤人肾小管上皮(HK-2)细胞的影响。方法:将HK-2细胞系分为正常对照组(Ctrl)、高糖组(HG)、甘露醇组(MA)和高糖+Ferrostatin-1组(HG+Fer-1)。试剂盒检测细胞内活性氧簇(ROS)、铁离子(Iron)、丙二醛(MDA)和谷胱甘肽(GSH)水平。透射电子显微镜观察细胞内线粒体形态学变化。qPCR和Western印迹检测溶质载体家族7成员11(SLC7A11)、谷氨酸-半胱氨酸连接酶催化亚基(GCLC)、谷胱甘肽过氧化物酶4(GPX4)、转铁蛋白受体1(TFR-1)mRNA和蛋白表达。结果:与Ctrl组相比,HG组细胞内ROS、Iron和MDA水平升高(F=17.72、14.33、39.53,均P<0.01),GSH含量下降(F=18.24,P<0.001);线粒体嵴断裂和膜密度增加;SLC7A11、GPX4和GCLC mRNA水平降低(F=22.22、19.43、22.3,均P<0.001),TFR-1 mRNA水平升高(F=10.01,P<0.01);SLC7A11、GPX4和GCLC蛋白水平降低(F=12.74、18.79、17.49,均P<0.01),TFR-1蛋白水平升高(F=15.08,P<0.01)。与HG组相比,HG+Fer-1组细胞内ROS、Iron和MDA水平降低(P<0.05、P<0.01、P<0.001),GSH含量增加(P<0.01);线粒体形态恢复正常;SLC7A11、GPX4和GCLC mRNA水平上调(P<0.01、P<0.01、P<0.001),TFR-1 mRNA水平下调(P<0.05);SLC7A11、GPX4和GCLC蛋白水平上调(均P<0.01),TFR-1蛋白水平下调(P<0.01)。结论:高糖诱导HK-2细胞铁死亡,促进细胞损伤。
Objective:To investigate the effect of ferroptosis on high-glucose injured human renal tubular epithelial(HK-2)cells.Methods:HK-2 cell line was divided into four groups:normal control group(Ctrl),high glucose group(HG),mannitol group(MA),and high glucose+Ferrostatin-1 group(HG+Fer-1).The reagent kit was used to detect intracellular levels of reactive oxygen species(ROS),iron ions(Iron),malondialdehyde(MDA),and glutathione(GSH).Transmission electron microscopy was performed to observe the morphological changes of intracellular mitochondria.qPCR and Western blotting were performed to detect the mRNA and protein expression of solute carrier family 7 member 11(SLC7A11),glutathione peroxidase4(GPX4),glutamate-cysteine ligase catalytic subunit(GCLC),and transferrin receptor 1(TFR-1).Results:Compared with the Ctrl group,the HG group exhibited increased levels of ROS,Iron and MDA(F=17.72,13.2,39.53,all P<0.01)and decreased level of GSH(F=18.24,P<0.001);mitochondrial cristae rupture and membrane density were increased;SLC7A11,GPX4 and GCLC mRNA levels were decreased(F=22.22,19.43,22.3,all P<0.001)and TFR-1 mRNA level was increased(F=10.01,P<0.01);SLC7A11,GPX4 and GCLC protein levels decreased(F=12.74,18.79,17.49,all P<0.01)and TFR-1 protein level was increased(F=15.08,P<0.01).Compared with the HG group,intracellular ROS,Iron and MDA levels were decreased in the HG+Fer-1 group(P<0.05,P<0.01,P<0.001),GSH content was increased(P<0.01);mitochondrial morphology was returned to normal;SLC7A11,GPX4 and GCLC mRNA levels were upregulated(P<0.01,P<0.01,P<0.001),and TFR-1 mRNA levels were down-regulated(P<0.05);SLC7A11,GPX4 and GCLC protein levels were up-regulated(all P<0.01),and TFR-1 protein levels were down-regulated(P<0.01).Conclusion:High glucose induces ferroptosis and promotes cell injury in HK-2 cells.
作者
曾晓娇
田玲
张景云
ZENG Xiaojiao;TIAN Ling;ZHANG Jingyun(NHC Key Laboratory of Hormones and Development,Tianjin Key Laboratory of Metabolic Diseases,Chu Hsien-I Memorial Hospital&Tianjin Institute of Endocrinology,Tianjin Medical University,Tianjin 300134,China)
出处
《天津医科大学学报》
2024年第3期239-244,共6页
Journal of Tianjin Medical University