中国血液净化

中国血液净化 ›› 2023, Vol. 22 ›› Issue (09): 679-684.doi: 10.3969/j.issn.1671-4091.2023.09.010

• 基础研究 • 上一篇    下一篇

Ferrostatin-1在高糖诱导的肾小管上皮细胞铁死亡中的作用

吴子瑜    林 艳    梁媛琦    郭太林   

  1. 350001 福州，1福建医科大学省立临床医学院 2 福建省立医院老年科
  • 收稿日期:2023-03-13 修回日期:2023-07-11 出版日期:2023-09-12 发布日期:2023-09-12
  • 通讯作者: 吴子瑜 E-mail:851266719@qq.com
  • 基金资助:
    福建省卫生健康委员会基金计划项目(2019-1-22)

Role of Ferrostatin-1 prevent high glucose-induced Ferroptosis of renal tubular epithelial cells

WU Zi-yu, LIN Yan, LIANG Yuan-qi, GUO Tai-lin   

  1. The Provincial Clinical Medical College of Fujian Medical University；2Department of Geriatric Medicine , Fujian Provincial Hospital, Fuzhou 350001, China
  • Received:2023-03-13 Revised:2023-07-11 Online:2023-09-12 Published:2023-09-12
  • Contact: 350001 福州，1福建医科大学省立临床医学院 2福建省立医院老年科 E-mail:851266719@qq.com

PDF

523

摘要： 目的 探讨Ferrostatin-1(Fer-1)在高糖诱导的肾小管上皮细胞铁死亡中的作用。 方法  ①24只DBA/2J小鼠被随机分为4组：正常对照组、Fer-1干预组、糖尿病(diabetes mellitus,DM)模型组、DM+Fer-1干预组，每组6只。根据小鼠体质量，连续5天以40 mg/kg新鲜制备的链脲佐菌素腹腔注射DBA/2J小鼠以构建DM小鼠模型。Fer-1干预组及DM+Fer-1干预组在造模后每天予腹腔注射Fer-1(2.5 μmol/kg)，对照组及DM组予等体积生理盐水腹腔注射，连续12周。通过Western印迹法评估谷胱甘肽过氧化物酶4(glutathione peroxidase 4，GPX4)和轻链亚基(solute carrier family 7 A11,SLC7A11)蛋白的表达水平，免疫组化检测肾脏组织内GPX4蛋白的表达定位情况。②用高糖(30 mmmol/L)刺激人肾小管上皮细胞(human proximal tubular epithelial cells，HK-2)，采用浓度200 nm的Fer-1干预细胞， Western印迹及免疫荧光染色检测肾小管上皮细胞的铁死亡信号通路相关蛋白的表达情况。 结果 与正常对照组相比，DM模型组小鼠肾脏组织的铁(Fe2+)、丙二醛(malondialdehyde，MDA)含量上调,谷胱甘肽(glutathione，GSH)含量降低(t=35.073、15.732、4.954；均P＜0.001)，铁死亡相关标志物GPX4和SLC7A11蛋白表达量上调(t=12.432、5.484；P=0.006、0.003)。与DM模型组相比，DM+Fer-1干预组小鼠肾脏组织的Fe2+、MDA含量降低，GSH含量增加(t=14.503、4.675、7.124；均P＜0.001)，铁死亡相关标志物GPX4和SLC7A11的蛋白表达水平上调(t=4.573、27.942；P=0.039、P＜0.001)。高糖可诱导体外培养的HK-2细胞铁死亡相关蛋白GPX4和SLC7A11表达减少(t=12.433、7.716；P=0.006、0.019)，使用Fer-1可显著增加GPX4和SLC7A11的蛋白表达水平(t=5.136、5.043；P=0.036、0.037)。 结论 高糖诱导的肾小管上皮细胞出现铁死亡，抑制铁死亡可能是防治糖尿病肾脏病进展的新策略。

关键词: 铁死亡, 糖尿病肾脏病, 肾小管上皮细胞, Ferrostatin-1

Abstract: Objective To investigate the role of Ferrostatin-1(Fer-1) in high glucose-induced ferroptosis of renal tubular epithelial cells. Method (1) 24 DBA/2J mice were randomly divided into 4 groups: normal control group, Fer-1 intervention group, DM model group, DM + Fer-1 intervention group with 6 mice in each group. According to the body weight of mice, DBA/2J mice were injected intraperitoneally with freshly prepared Streptozocin at a dose of 40 mg / kg for 5 days to establish DM model. Fer-1 intervention group and DM+Fer-1 intervention group were given intraperitoneal injection of Fer-1 (2.5μmol/kg) every day for 12 weeks after modeling.. The control group and DM Group were intraperitoneally injected with the same volume of normal saline for 12 weeks. The expression levels of activated glutathione peroxidase 4(GPX4)and solute carrier family 7 A11(SLC7A11)were detected by Western blotting, and the expression and localization of GPX4 in renal tissue were detected by immunohistochemistry.(2) Human renal tubular epithelial cells (HK-2) were stimulated with high glucose (30 mmol/L) , and treated with 200 nm Fer-1. The expression of ferroptosis signaling in renal tubular epithelial cells was detected by Western blotting and immunofluorescence staining. Results  Compared with the normal control group, the levels of Fe2+and Malondialdehyde (MDA) in the kidney of DM model group were significantly up-regulated, glutathione (GSH) was significantly down-regulated ( t=35.073、15.732 and 4.954 respectively，all P ＜ 0.001) , and the protein expression levels of GPX4 and SLC7A11, which were related to ferroptosis, were significantly up-regulated (t=12.432 and 5.484；P=0.006 and 0.003) . Compared with the DM model group, the levels of Fe2+ and MDA in the kidney of DM +Fer-1 intervention group were down-regulated, while GSH was up-regulated (t=14.503、4.675 and 7.124； all P＜0.001) . The protein expression levels of GPX4 and SLC7A11 were up-regulated (t =4.573 and 27.942；P= 0.039 and 0.001) . In addition, high glucose could induce the expression levels of ferroptosis related proteins GPX4 and SLC7A11 in HK-2 cells cultured in vitro to decrease significantly ( t=12.433 and 7.716；P=0.006 and 0.019) , the expression levels of GPX4 and SLC7A11 were significantly increased by Fer-1(t=5.136 and 5.043；P=0.035 and 0.037) . Conclusions  Inhibition of ferroptosis in renal tubular epithelial cells induced by high glucose may be a new strategy to prevent the progression of DM.

Key words: Ferroptosis, Diabetic kidney disease, Renal tubular epithelial cells, Ferrostatin-1

中图分类号: