脱氢胆酸调控OPG/RANK和TRAF3抑制破骨细胞分化  

Dehydrocholic acid inhibits osteoclast differentiation by regulating OPG/RANK and TRAF3

作　　者：朱禹潼 张晓楠 关溪 尚东[1,2,3] ZHU Yutong;ZHANG Xiaonan;GUAN Xi;SHANG Dong(Laboratory of Integrative Medicine,the First Affiliated Hospital of Dalian Medical University,Dalian 116011,Liaoning;Institute(College)of Integrative Medicine,Dalian Medical University,Dalian 116044,Liaoning;Department of General Surgery,the First Affiliated Hospital of Dalian Medical University,Dalian 116011,Liaoning,China)

机构地区：[1]大连医科大学附属第一医院中西医结合临床重点学科实验室,辽宁大连116011 [2]大连医科大学中西医结合研究院(学院),辽宁大连116044 [3]大连医科大学附属第一医院普外三科,辽宁大连116011

出　　处：《中国骨质疏松杂志》2024年第1期12-16,共5页Chinese Journal of Osteoporosis

基　　金：国家重点研发计划(2018YFE0195200)。

摘　　要：目的探讨脱氢胆酸(dehydrocholic acid,DHCA)对破骨细胞(osteoclasts,OCs)分化及功能的影响。方法采用粒细胞-巨噬细胞集落刺激因子和核因子κB受体活化因子配体(receptor activator of nuclear factor kappa B(NF-κB)-ligand,RANKL)诱导成熟骨髓来源的巨噬细胞分化为OCs。通过抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP/ACP5)染色,确定DHCA抑制OCs形成的最佳浓度。qRT-PCR检测OCs分化和功能相关基因TRAP/ACP5、组织蛋白酶K(cathepsin K,CTSK)和基质金属蛋白酶9(matrix metalloproteinase 9,MMP9)的基因表达。Western blot检测OCs中TNF受体相关因子3(TNF receptor-associated factor 3,TRAF3)、NF-κB受体活化因子(receptor activator of NF-κB,RANK)和骨保护素(osteoprotegerin,OPG)的蛋白水平。结果DHCA浓度为200μmol/L是抑制OCs形成的最佳剂量(P<0.01)。DHCA抑制OCs分化和功能相关基因ACP5、CTSK和MMP9的表达(P<0.01)。DHCA通过下调RANK蛋白和增加TRAF3和OPG蛋白的表达来抑制OCs的分化(P<0.01)。结论DHCA通过调控OPG/RANK和TRAF3抑制OCs分化,这可能是一种有效的骨质疏松前体药物。Objective To investigated the effect of dehydrocholic acid(DHCA)on osteoclast(OCs)differentiation and function.Methods Granulocyte macrophage colony-stimulating factor and receptor activator of nuclear factor kappa B(NF-κB)-ligand(RANKL)were used to differentiate mature bone marrow macrophages into OCs.To determine the best concentration of DHCA to inhibit OC formation,tartrate resistant acid phosphatase(TRAP)staining was conducted.The gene expressions of tartrate-resistant acid phosphatase(TRAP/ACP5),cathepsin K(CTSK),matrix metalloproteinase 9(MMP9),and OC differentiation-and function-related genes were detected using qRT-PCR.The protein levels of TNF receptor-associated factor 3(TRAF3),receptor activator of NF-κB(RANK),and osteoprotegerin(OPG)in OCs were measured with Western blotting.Results DHCA at a concentration of 200μmol/L was the optimal dose to inhibit OC formation in vitro(P<0.01).DHCA inhibited the differentiation-and function-related genes ACP5,CTSK,and MMP9 of OCs(P<0.01).Mechanically,DHCA inhibited OC differentiation by down-regulating RANK proteins and increasing the expression of TRAF3 and OPG proteins(P<0.01).Conclusion DHCA inhibits OC differentiation by regulating OPG/RANK and TRAF3.This may be an effective prodrug for anti-osteoporosis.

关 键 词：脱氢胆酸 破骨细胞 OPG/RANK TRAF3 骨质疏松 

分 类 号：R683[医药卫生—骨科学] R336[医药卫生—外科学]