脉冲磁疗对成骨细胞铁死亡的影响  

Effects of pulsed magnetic field therapy on osteoblasts to resist ferroptosis

作　　者：丁奕栋 刘功稳 张辉 王爱飞[4] 魏祺 郑苗 徐又佳[4] DING Yi-dong;LIU Gong-wen;ZHANG Hui;WANG Ai-fei;WEI Qi;ZHENG Miao;XU You-jia(Clinical College,Suzhou Vocational Health College,Suzhou 215000,Jiangsu,China;Department of Orthopedics,Suzhou Hospital of Traditional Chinese Medicine,Suzhou 215000,Jiangsu,China;Department of Orthopedics,Anhui Provincial Hospital,Hefei 230001,China;Department of Orthopedics,the Second Affiliated Hospital of Soochow University,Suzhou 215004,Jiangsu,China)

机构地区：[1]苏州卫生职业技术学院临床医学院,江苏苏州215000 [2]苏州市中医院骨科,江苏苏州215000 [3]安徽省省立医院骨科,合肥230001 [4]苏州大学附属第二医院骨科,骨质疏松症临床中心,苏州大学骨质疏松诊疗技术研究所,江苏苏州215004

出　　处：《中华骨质疏松和骨矿盐疾病杂志》2022年第3期264-269,共6页Chinese Journal Of Osteoporosis And Bone Mineral Research

基　　金：国家重点研发计划(2021YFC2501702);国家自然科学基金(82072474);江苏省体医融合促进老年骨骼健康应用工程研究中心。

摘　　要：目的从细胞水平探索脉冲磁疗对成骨细胞保护和抵抗铁死亡作用的机制。方法将成骨细胞用低频脉冲磁疗仪进行30 min的干预。实验组分为6组:对照(control,Cont)组、磁疗(magnetic therapy,MT)组、Erastin(3 mmol/L)铁死亡诱导(Erastin ferroptosis induced,Era)组、Era+MT组、铁死亡诱导+GW4869外泌体抑制(1.5 mmol/L)(Era+GW)组、Era+MT+GW组。Western blot法检测铁死亡相关蛋白谷胱甘肽过氧化酶4(glutathione peroxidase 4,GPX4)、外泌体表面标记蛋白(CD9,CD63)、外泌体内铁蛋白(Ferritin)。扫描电镜检测成骨细胞外泌体。结果Western blot法检测结果表明,磁疗治疗后成骨细胞GPX4得到提升,MT组高于Cont组(1.34±0.01 vs.1.00±0.01,t=200.2,P<0.05),Era+MT组高于Era组(0.87±0.08 vs.0.77±0.06,t=1.12,P<0.05),差异有统计学意义。MT组相对Cont组[(3.57±0.01)μmol/L vs.(3.54±0.03)μmol/L,t=2.89,P<0.05],Era+MT组相对Era组[(5.63±0.63)μmol/L vs.(3.59±0.05)μmol/L,t=3.23,P<0.05],培养基中铁离子浓度上升;并且Western bot法检测发现外泌体铁蛋白含量MT组高于Cont组(2.42±0.09 vs.1.01±0.01),差异有统计学意义(t=14.69,P<0.05)。GW4869抑制外泌体后,以上指标改变不明显。结论磁疗减少成骨细胞的铁死亡,挽救成骨细胞铁死亡的机制由一个“磁疗-外泌体-铁蛋白”途径驱动,并对铁稳态、细胞内运输和骨质疏松症有广泛的影响。Objective To explore the mechanism of magnetic therapy to protect osteoblasts from ferroptosis.Methods Osteoblasts were treated with a magnetotherapy machine for 30 minutes.The experimental group was divided into 6 groups:control(Cont)group,magnetic therapy(MT)group,Erastin(3 mmol/L)ferroptosis induced(Era)group,ferroptosis induced with magnetic therapy(Era+MT)group,ferroptosis induced with GW4869 exosome inhibited(1.5 mmol/L)(Era+GW)group,ferroptosis induced with magnetic therapy and exosome inhibited(Era+MT+GW)group.Western blot was used to detect ferroptosis related proteins glutathione peroxidase 4(GPX4),exosomal surface marker proteins(CD9,CD63)expression.Osteoblastic exosomes were detected by scanning electron microscope.Results Western blot showed that after magnetic therapy,GPX4 in MT group was significantly enhanced compared with Cont group(1.34±0.01 vs.1.00±0.01,t=200.2,P<0.05),iron in the medium of MT group was significantly higher than Cont group[(3.57±0.01)μmol/L vs.(3.54±0.03)μmol/L,t=2.89,P<0.05].GPX4 protein in Era+MT group was significantly higher than Era group(0.87±0.08 vs.0.77±0.06,t=1.12,P<0.05)and iron ion in medium of Era+MT group was significantly higher than Era group(5.63±0.63μmol/L vs.3.59±0.05μmol/L,t=3.23,P<0.05).Western blot showed exosomal Ferritin in MT group was higer than Cont group(2.42±0.09 vs.1.01±0.01,t=14.69,P<0.05).But these effects in GW4869 inhibiting exosomes group were not obvious.Conclusion Magnetic therapy promotes the function of osteoblasts and the mechanism of saving the ferroptosis of osteoblasts can be driven by a magnetotherapy-exosome-ferritin pathway,which broad implications for iron homeostasis,intracellular trafficking,and osteoporosis.

关 键 词：脉冲磁场疗法 外泌体 铁死亡 成骨细胞 

分 类 号：R681[医药卫生—骨科学]