盐酸戊乙奎醚抑制亚铁肌红蛋白诱导的人近端肾小管上皮细胞铁死亡  

Penehyclidine Hydrochloride Inhibits Ferrous Myoglobin Induced Ferroptosis of Human Renal Cortex Proximal Tubule Epithelial Cells

作　　者：罗莎莎 陈莉 王东伟 谭红保[1] LUO Shasha;CHEN Li;WANG Dongwei(Department of Anesthesiology,The Fourth Hospital of Changsha,Hunan 410006,China)

机构地区：[1]长沙市第四医院麻醉科,410006 [2]湖南省脑科医院(湖南省第二人民医院)肾内科,长沙410007

出　　处：《医学研究杂志》2024年第4期91-96,共6页Journal of Medical Research

基　　金：湖南省自然科学基金资助项目(2020JJ8039);湖南省长沙市自然科学基金资助项目(Kq2202485)。

摘　　要：目的 探讨盐酸戊乙奎醚(penehyclidine hydrochloride, PHC)对亚铁肌红蛋白诱导人近端肾小管上皮(human renal cortex proximal tubule epithelial, HK2)细胞铁死亡的作用及其机制。方法 采用6mg/ml亚铁肌红蛋白处理HK2细胞建立横纹肌溶解(rhabdomyolysis, RM)致急性肾损伤(acute kidney injury, AKI)细胞模型。加入PHC与亚铁肌红蛋白共同孵育HK2细胞,然后使用铁抑素-1(Fer-1,铁死亡抑制剂)和Erastin(铁死亡诱导剂)作为实验干预。CCK8法检测细胞活性,试剂盒法检测亚铁离子(Fe2+)和丙二醛(malondialdehyde, MDA)水平,流式细胞术检测脂质活性氧(reactive oxygen species, ROS)和线粒体膜电位(mitochondrial membrane potential, MMP)水平,实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction, RT-qPCR)和Western bolt法检测GPX4的蛋白和mRNA表达水平。结果 PHC处理显著增加了细胞活性,同时降低了Fe2+、ROS和MDA水平,增加了MMP水平。与PHC单独处理比较,加入Fer-1后可以进一步增加细胞活性,下调Fe2+水平,降低ROS和MDA水平,升高MMP水平。而加入Erastin后,结果刚好相反。此外,从机制上讲,PHC处理上调了SLC7A11和GPX4水平。结论 PHC可以通过抑制铁死亡保护HK2细胞免受亚铁肌红蛋白损伤,这可能与SLC7A11/GPX4通路有关。Objective To explore the role and mechanism of penehyclidine hydrochloride(PHC)on ferroptosis induced by ferrous myoglobin in human renal cortex proximal tubule epithelial(HK2)cells.Methods HK2 cells were treated with 6mg/ml ferrous myoglobin to establish rhabdomyolysis(RM)induced acute kidney injury(AKI)cell models.PHC and ferrous myoglobin were added to incubate the cells.The ferrostatin-1(Fer-1,ferroptosis inhibitor)and Erastin(ferroptosis inducer)were then employed as experimental interventions.The cell activity was determined by CCK-8method,and the level of Fe 2+and MDA were detected by assay kits,the level of lipid reactive oxygen species(ROS)and mitochondrial membrane potential(MMP)were detected by flow cytometry.The protein and mRNA expression levels of GPX4 were detected by real-time quantitative polymerase chain reaction(RT-qPCR)and Western bolt(WB).Results PHC treatment significantly increased the cell activity,decreased the levels of Fe 2+,ROS,MDA.and increased the level of MMP.Compared with PHC alone treatment,the addition of Fer-1 increased cell activity further,down-regulated the level of Fe 2+,decreased the levels of ROS and MDA,and increased the level of MMP.After adding Erastin,the results were be opposited.In addition,PHC treatment mechanistically up-regulated the levels of SLC7A11 and GPX4.Conclusion PHC can protect HK2 cells from ferromyoglobin damage by inhibiting ferroptosis,which may be related to the SLC7A11/GPX4 pathway.

关 键 词：横纹肌溶解 急性肾损伤 铁死亡 盐酸戊乙奎醚 SLC7A11/GPX4通路 

分 类 号：R692[医药卫生—泌尿科学]