金雀根通过阻断TRPM7抑制Erastin诱导的软骨细胞铁死亡
Caragana sinica root inhibits Erastin-induced chondrocyte ferroptosis by blocking TRPM7
朱仁弟 1屈彪 2周仁鹏 1胡伟1
作者信息
- 1. 安徽医科大学药学院,合肥 230032,安徽;安徽医科大学第二附属医院药物临床试验研究中心,合肥 230601,安徽
- 2. 安徽医科大学第二附属医院药物临床试验研究中心,合肥 230601,安徽
- 折叠
摘要
目的:探讨金雀根(Caragana sinica root,CSR)对Erastin诱导的软骨细胞铁死亡模型中铁死亡的影响及可能的机制.方法:培养C28/I2软骨细胞系,构建Erastin诱导铁死亡的细胞模型.采用MTT法检测细胞活力;Calcein/PI染色观察细胞活性;通过试剂盒检测细胞乳酸脱氢酶(LDH)及总谷胱甘肽(GSH)水平;荧光探针BODIPY 581/591 C11标记检测活性氧(ROS)水平;Rh123和JC-1染色观察线粒体膜电位(ΔΨm)的变化;Western blot检测铁死亡相关蛋白(ACSL4、GPX4)和TRPM7蛋白的表达.结果:Erastin处理可降低软骨细胞活力,增加细胞毒性,诱发氧化应激,破坏ΔΨm,并上调ACSL4蛋白表达,同时下调GPX4蛋白表达,诱导软骨细胞铁死亡.相反,金雀根可以使细胞活力恢复,降低氧化应激反应,从而抑制软骨细胞铁死亡.此外,金雀根能降低Erastin引起的TRPM7蛋白表达水平的升高.结论:Erastin诱发了 C28/I2软骨细胞脂质过氧化反应,引起线粒体损伤及铁死亡;金雀根可能通过阻断TRPM7抑制软骨细胞铁死亡,从而发挥保护软骨细胞的作用.
Abstract
AIM:To investigate the effect of Cara-gana sinica root(CSR)on ferroptosis in the Erastin-induced chondrocyte ferroptosis model and possi-ble mechanisms.METHODS:The C28/I2 chondro-cyte cell line was cultured to construct a cell model of Erastin-induced ferroptosis.Cell viability was de-tected by MTT assay;cell death was observed by Calcein/PI staining;cell lactate dehydrogenase(LDH)and total glutathione(GSH)levels were de-tected by the kit;reactive oxygen species(ROS)lev-els were detected by fluorescent probe BODIPY 581/591 C11 labeling;mitochondrial membrane po-tential(ΔΨm)changes were observed by Rh123 and JC-1 staining;Western blot was used to detect the expression of ferroptosis-related proteins(AC-SL4,GPX4)and TRPM7 proteins.RESULTS:Erastin treatment decreased chondrocyte viability,in-creased cytotoxicity,induced oxidative stress,dis-rupted ΔΨm,and up-regulated ACSL4 protein ex-pression,while down-regulating GPX4 protein ex-pression and inducing chondrocyte ferroptosis.In contrast,CSR restored cell viability and reduced oxi-dative stress,thereby inhibiting chondrocyte fer-roptosis.In addition,CSR reduced the Erastin-in-duced increase in TRPM7 protein expression level.CONCLUSION:Erastin induced lipid peroxidation in C28/I2 chondrocytes,causing mitochondrial dam-age and ferroptosis;CSR may inhibit chondrocyte ferroptosis by blocking TRPM7,thus exerting a pro-tective effect on chondrocytes.
关键词
金雀根/软骨细胞/Erastin/铁死亡/TRPM7Key words
Caragana sinica root/chondrocytes/Erastin/ferroptosis/TRPM7引用本文复制引用
基金项目
国家自然科学基金(81902182)
国家自然科学基金(82071591)
国家自然科学基金(82371575)
出版年
2024
国家科技期刊平台
NSTL
万方数据