Apelin-13通过Nrf2-焦亡通路抑制RANKL诱导的巨噬细胞向破骨细胞分化  

Apelin-13 inhibits RANKL-induced macrophage differentiation into osteoclasts via the Nrf2-pyroptosis pathway

作　　者：王超[1,2,3,4] 邹秀强 刘晓 殷照阳 WANG Chao;ZOU Xiuqiang;LIU Xiao;YIN Zhaoyang(Lianyungang Clinical College of Nanjing Medical University(The First People’s Hospital of Lianyungang),Lianyungang 222000,China;Department of Orthopedics,the Affiliated Jiangning Hospital with Nanjing Medical University,Nanjing 211100,China;Department of Orthopedics,Jiangning Clinical College of Medicine,Kangda College,Nanjing Medical University,Nanjing 211100,China;Department of Orthopedics,Jiangning Clinical Hospital of Jiangsu Health Vocational College,Nanjing 211100,China;Department of Orthopedics,the Affiliated Lianyungang Hospital of Xuzhou Medical University(The First People’s Hospital of Lianyungang),Lianyungang 222000,China)

机构地区：[1]南京医科大学连云港临床医学院(连云港市第一人民医院),江苏连云港222000 [2]南京医科大学附属江宁医院骨科,江苏南京211100 [3]南京医科大学康达学院江宁临床医学院骨科,江苏南京211100 [4]江苏卫生健康职业学院江宁临床医院,江苏南京211100 [5]徐州医科大学附属连云港医院(连云港市第一人民医院),江苏连云港222000

出　　处：《中国骨质疏松杂志》2025年第3期343-348,408,共7页Chinese Journal of Osteoporosis

基　　金：江苏省卫健委科研项目(LGY2020052、2022-208、Z2022086);南京市卫生科技发展专项资金项目(重点项目,ZKX22061);连云港市科技局项目(SF2118);连云港市卫健委面上项目(202212);南京医科大学康达学院科研发展基金(KD2023KYJJ242);江苏卫生健康职业学院科研项目(JKC2022052)。

摘　　要：目的关节假体磨损颗粒诱发骨溶解是导致关节置换术后无菌性松动的主要原因。Apelin-13是具有抗炎作用的脂肪因子家族成员。本文拟探讨Apelin-13通过Nrf2-焦亡通路抑制巨噬细胞向破骨细胞分化的分子机制。方法RANKL刺激RAW264.7小鼠巨噬细胞,同时加入Apelin-13和Nrf2抑制剂ML385。将细胞分为4组,分别为Sham组,RANKL组,RANKL+Apelin-13组(Apelin-13组)和RANKL+Apelin-13+ML385组(ML385组)。CCK8法检测细胞活力,TRAP染色法检测破骨细胞的生成,ELISA法检测IL-1β和IL-18的水平,Western blot检测焦亡蛋白和抗氧化应激蛋白的表达水平。结果0.1、1、10 nmol/L和25 nmol/L剂量的Apelin-13对巨噬细胞的存活率没有显著影响。Apelin-13显著抑制了RAW264.7向破骨细胞分化,且呈剂量依赖性。Apelin-13增加了抗氧化应激蛋白表达,并抑制了焦亡蛋白水平,这一保护效应被ML385所抵消。结论Apelin-13通过Nrf2途径抑制RAW264.7细胞的焦亡和破骨细胞的形成。Objective Osteolysis,triggered by wear particles from joint prostheses,is a primary factor leading to aseptic loosening after joint replacement surgery.Apelin-13,a member of the adipokine family known for its anti-inflammatory properties,is investigated in this study for its role in inhibiting macrophage differentiation into osteoclasts via the Nrf2-pyroptosis pathway.Methods RAW264.7 mouse macrophages were stimulated by RANKL in the presence of Apelin-13 and the Nrf2 inhibitor ML385.The cells were then categorized into four groups:Sham group,RANKL group,RANKL+Apelin-13 group(Apelin-13 group),and RANKL+Apelin-13+ML385 group(ML385 group).Cell viability was assessed using the CCK8 method.Osteoclast generation was evaluated through TRAP staining.Levels of IL-1βand IL-18 were measured using ELISA.The expression levels of pyroptosis and antioxidant stress proteins were analyzed with Western blotting.Results Apelin-13 did not have a significant impact on macrophage survival at doses of 0.1,1,10 nmol/L,and 25 nmol/L.However,it did show a dose-dependent inhibition of RAW264.7 differentiation into osteoclasts.Additionally,Apelin-13 increased the expression of anti-oxidative stress proteins and decreased pyroptosis protein levels,and this protective effect was reversed by ML385.Conclusion Apelin-13 inhibits pyroptosis and osteoclast formation in RAW264.7 cells through the Nrf2 pathway.

关 键 词：APELIN-13 假体周围骨溶解 破骨细胞 NRF2 焦亡 

分 类 号：R684[医药卫生—骨科学]