异泽兰黄素通过调控MAPK和NF-κB信号通路抑制RANKL诱导的破骨细胞分化  

Eupatilin Inhibits RANKL-induced Osteoclast Differentiation by Regulating MAPK and NF-κB Signaling Pathways

作　　者：赵博轩 李婷 姚汉涛 王紫君 郭海盈 季耀庭 杜民权[1] ZHAO Boxuan;LI Ting;YAO Hantao;WANG Zijun;GUO Haiying;JI Yaoting;DU Minquan(State Key Laboratory of Oral&Maxillofacial Reconstruction and Regeneration,Key Laboratory of Oral Biomedicine Ministry of Education,Hubei Key Laboratory of Stomatology,School&Hospital of Stomatology,Wuhan University,Wuhan 430079,China)

机构地区：[1]口颌系统重建与再生全国重点实验室,口腔生物医学教育部重点实验室,口腔医学湖北省重点实验室,武汉大学口腔医(学)院,湖北武汉430079

出　　处：《口腔医学研究》2024年第9期820-826,共7页Journal of Oral Science Research

基　　金：国家自然科学基金(编号:82172493、82372463、81771084、81700772)。

摘　　要：目的:探究异泽兰黄素(Eupatilin)对破骨细胞前体细胞向破骨细胞分化的作用和分子机制。方法:CCK-8法检测不同浓度的Eupatilin对破骨细胞前体细胞RAW264.7以及小鼠骨髓巨噬细胞(bone marrow-derived macrophages,BMDMs)细胞活性的影响。用核因子κB受体活化因子配体(RANKL)诱导RAW264.7和BMDMs分化形成破骨细胞,同时使用不同浓度的Eupatilin(5、10、20μmol/L)干预,通过TRAP染色和F-actin染色评价Eupatilin对RANKL诱导的破骨细胞形成作用,qRT-PCR和Western blot检测破骨细胞相关标志基因的表达,Western blot检测丝裂原活化蛋白激酶(MAPK)和核因子-κB(NF-κB)信号通路分子的磷酸化水平。结果:20μmol/L Eupatilin对破骨细胞的分化具有显著的抑制作用,同时对相关标志基因抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphase,TRAP)、基质金属蛋白酶-9(matrix metallopeptidase-9,MMP-9)、组织蛋白酶K(cathepsin K,CTSK)等的表达也表现出有效的下调作用。Western blot结果显示Eupatilin显著抑制了RANKL诱导的MAPK和NF-κB信号通路分子的磷酸化水平。结论:Eupatilin通过调控MAPK和NF-κB信号通路,对RANKL诱导的破骨细胞分化具有显著的抑制作用。Objective:To investigate the effects and molecular mechanisms of eupatilin on the differentiation of osteoclast precursor cells to osteoblasts.Methods:CCK-8 assay was used to detect the effects of different concentrations of Eupatilin on the cell activity of osteoclast precursor cells RAW264.7 and mouse bone marrow-derived macrophages(BMDMs).RAW264.7 and BMDMs were induced to differentiate into osteoclasts by receptor activator of nuclear factor-κB ligand(RANKL),and different concentrations of Eupatilin(5,10,and 20μmol/L)were used for intervention.The effects of Eupatilin on RANKL-induced osteoclast formation were evaluated through TRAP staining and F-actin staining.The expression of osteoclast-related marker genes was detected by qRT-PCR and Western blot,and the phosphorylation levels of mitogen-activated protein kinase(MAPK)and nuclear factor-κB(NF-κB)signaling pathway molecules were detected by Western blot.Results:Eupatilin at 20μmol/L exhibited a significant inhibitory effect on the differentiation of osteoclasts,and also shown an effective downregulation of the expression of related marker genes such as tartrate-resistant acid phosphas(TRAP),matrix metalloproteinase-9(MMP-9),and cathepsin K(CTSK).Western blot results indicated that Eupatilin significantly inhibited the phosphorylation levels of MAPK and NF-κB signaling pathway molecules induced by RANKL.Conclusion:Eupatilin exerts a significant inhibitory effect on RANKL-induced osteoclast differentiation by regulating the MAPK and NF-κB signaling pathways.

关 键 词：异泽兰黄素 破骨细胞 MAPK信号通路 NF-ΚB信号通路 

分 类 号：R781.42[医药卫生—口腔医学]