槲皮素对Erastin诱导的软骨细胞铁死亡的影响  

Effect of quercetin on Erastin-induced ferroptosis in chondrocytes

作　　者：王浩 周富丽 朱仁弟 赵英杰 周仁鹏 胡伟[2] 鲁超[1,3] WANG Hao;ZHOU Fu-li;ZHU Ren-di;ZHAO Ying-jie;ZHOU Ren-peng;HU Wei;LU Chao(School of Pharmacy,Anhui Medical University,Hefei 230032,China;Dept of Clinical Pharmacology,the Second Hospital of Anhui Medical University,Hefei 230601,China;the First Affiliated Hospital of Anhui University of Science&Technology,Huainan Anhui 232001,China)

机构地区：[1]安徽医科大学药学院,安徽合肥230032 [2]安徽医科大学第二附属医院药物临床试验研究中心,安徽合肥230601 [3]安徽理工大学第一附属医院,安徽淮南232001

出　　处：《中国药理学通报》2024年第10期1945-1952,共8页Chinese Pharmacological Bulletin

基　　金：安徽省自然科学基金资助项目(No 2208085MH215);安徽省中医药传承创新科研项目(No 2020CCZD05);安徽省重点研究与开发计划项目(No 202204295107020035)。

摘　　要：目的探究槲皮素(quercetin,Que)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。方法使用Erastin诱导C28/I2软骨细胞铁死亡模型。采用不同浓度的Que处理C28/I2软骨细胞,分别用MTT法检测细胞活性,LDH法检测细胞毒性。Western blot检测软骨细胞Prdx6和铁死亡相关蛋白ACSL4、GPX4的表达水平。流式细胞术检测软骨细胞脂质ROS的产生情况。RH123染色法检测软骨细胞线粒体膜电位的变化情况。RT-qPCR检测软骨细胞Prdx6 mRNA表达水平。免疫荧光染色法观察铁死亡相关蛋白ACSL4、GPX4的表达变化情况。结果与Erastin诱导的铁死亡模型组相比,Que可以明显提高C28/I2软骨细胞活性,降低细胞毒性。降低铁死亡相关蛋白ACSL4的表达,升高GPX4的表达。同时抑制软骨细胞脂质ROS的产生,并增强其线粒体膜电位。此外,与Control组相比,Erastin组Prdx6的表达明显降低,而使用Que可以上调Prdx6的表达。同时,在使用Prdx6的抑制剂MJ33后,Que对软骨细胞铁死亡的抑制作用下降。结论Que可以抑制Erastin诱导的C28/I2软骨细胞铁死亡,并可能通过上调Prdx6抑制软骨细胞铁死亡,从而发挥保护软骨细胞的作用。Aim To explore the effect of quercetin(Que)on ferroptosis and the potential mechanisms in an Erastin-induced ferroptosis model in chondrocytes.Methods A model of Erastin-induced ferroptosis was established in C28/I2 chondrocytes.Cells were treated with different concentrations of Que.Cell viability and cytotoxicity were assessed by MTT and LDH assays.The expression levels of Prdx6 and ferroptosis-related proteins ACSL4 and GPX4 in chondrocytes were determined by Western blot.Lipid ROS production in chondrocytes was measured by flow cytometry,while the changes in mitochondrial membrane potential were detected by RH123 staining.Prdx6 mRNA expression in chondrocytes was quantified by RT-qPCR.The changes in the expression of the ferroptosis-related proteins ACSL4 and GPX4 were detected by immunofluorescence staining.Results Compared to the Erastin-induced ferroptosis model group,Que significantly improved the viability of C28/I2 chondrocytes and reduced cell cytotoxicity.It decreased the expression of the ferroptosis-related protein ACSL4 and increased the expression of GPX4.Que also inhibited the production of lipid ROS in chondrocytes and strengthened their mitochondrial membrane potential.In addition,the expression of Prdx6 was significantly reduced in the Erastin group compared to the control group,while Que treatment upregulated the expression of Prdx6.Meanwhile,the inhibitory effect of Que on chondrocyte ferroptosis was reduced by the use of MJ33,an inhibitor of Prdx6.Conclusion Que can inhibit Erastin-induced ferroptosis of C28/I2 chondrocytes,possibly by upregulating Prdx6,and thus play a protective role in chondrocytes.

关 键 词：Prdx6 ROS Erastin QUE 铁死亡 软骨细胞 

分 类 号：R285.5[医药卫生—中药学] R329.24[医药卫生—中医学] R345.4R591.1R684.3