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
朱禹潼 1张晓楠 1关溪 2尚东3
作者信息
- 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;大连医科大学中西医结合研究院(学院),辽宁 大连 116044
- 2. 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
- 3. 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;大连医科大学中西医结合研究院(学院),辽宁 大连 116044;大连医科大学附属第一医院普外三科,辽宁 大连 116011
- 折叠
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 200 μmol/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.
Abstract
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 200 μmol/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Key words
dehydrocholic acid/osteoclasts/OPG/RANK/TRAF3/osteoporosis引用本文复制引用
基金项目
国家重点研发计划(2018YFE0195200)
出版年
2024
NETL
NSTL
维普
万方数据