BRD4对钛颗粒诱导破骨细胞形成和骨吸收功能作用  

ROLE OF BROMODOMAIN-CONTAINING PROTEIN 4 IN TITANIUM PARTICLE-INDUCED OSTEOCLASTOGENESIS AND BONE RESORPTION

作　　者：赵志平 王向宇[1,2] 贾斌 许应星 王昌耀 王英振[2] ZHAO Zhiping;WANG Xiangyu;JIA Bin;XU Yingxing;WANG Changyao;WANG Yingzhen(Department of Medicine,Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学医学部,山东青岛2660711 [2]青岛大学附属医院关节外科,山东青岛266071

出　　处：《青岛大学学报（医学版）》2022年第4期489-493,共5页Journal of Qingdao University(Medical Sciences)

基　　金：国家自然科学基金面上项目(81772329)。

摘　　要：目的研究表观遗传信号分子-溴结构域蛋白4(BRD4)在钛(Ti)颗粒体外诱导破骨细胞形成和骨吸收功能中的作用。方法小鼠RAW264.7巨噬细胞随机分为3组,对照组应用细胞培养液+100μg/L核因子κB受体活化因子配体(RANKL)+50μg/L巨噬细胞集落刺激因子(M-CSF)培养,Ti组应用细胞培养液+0.1g/L Ti+100μg/LRANKL+50μg/L M-CSF培养,Ti+JQ1组应用细胞培养液+0.1g/LTi+100μg/LRANKL+50μg/L M-CSF+200nmol/LJQ1培养。各组细胞培养48h后,应用CCK8检测细胞活性,荧光定量聚合酶链式反应法检测BRD4基因和破骨细胞相关基因组织蛋白酶K(CK)、抗酒石酸酸性磷酸酶(TRAP)、肿瘤坏死因子受体相关因子6(TRAF6)、活化T细胞核因子1(NFATc1)等mRNA表达;培养5d至破骨细胞形成,用TRAP染色法检测各组成熟破骨细胞数量,骨吸收培养板(OAP)检测各组破骨细胞骨吸收功能。结果各组细胞活性差异无显著性(F=1.629,P>0.05);Ti组BRD4基因和破骨细胞相关基因mRNA表达均较对照组显著升高,Ti+JQ1组BRD4基因和破骨细胞相关基因mRNA表达均较Ti组降低,差异有统计学意义(F=24.575~336.704,P<0.05);Ti组TRAP染色阳性的破骨细胞数目较其他两组明显增多,差异有统计学意义(F=165.941,P<0.05);Ti组骨吸收面积较对照组增加,Ti+JQ1组骨吸收面积与Ti组相比明显减少,差异均有统计学意义(F=49.879,P<0.05)。结论Ti颗粒通过上调RAW264.7巨噬细胞中BRD4蛋白的表达,促进破骨细胞形成和骨吸收功能。Objective To investigate the role of the epigenetic signaling molecule bromodomain-containing protein 4(BRD4)in titanium(Ti)particle-induced osteoclastogenesis and bone resorption in vitro.Methods Mouse RAW264.7 macrophages were randomly divided into control group,Ti group,and Ti+JQ1 group.The macrophages in the control group were cultured with cell culture medium+100μg/L receptor activator of nuclear factor-κB ligand(RANKL)+50μg/L macrophage colony-stimulating factor(M-CSF),those in the Ti group were cultured with cell culture medium+0.1 g/L Ti+100μg/L RANKL+50μg/L M-CSF,and those in the Ti+JQ1 group were cultured with cell culture medium+0.1 g/L Ti+100μg/L RANKL+50μg/L M-CSF+200 nmol/L JQ1.After culture for 48 h,CCK8 assay was used to measure cell viability,and quantitative real-time PCR was used to measure the mRNA expression ofBRD4gene and osteoclast-related genes such as cathepsin K(CK),tartrate-resistant acid phosphatase(TRAP),TNF receptor-associated factor 6(TRAF6),and nuclear factor of activated T-cells 1(NFATc1);after culture for 5 d till the formation of osteoclasts,TRAP staining was used to measure the number of mature os-teoclasts in each group,and osteo assay plate(OAP)was used to evaluate osteoclast-mediated bone resorption in each group.Results There was no significant difference in cell viability between groups(F=1.629,P>0.05).Compared with the control group,the Ti group had significantly higher mRNA expression levels ofBRD4gene and osteoclast-related genes,and compared with the Ti group,the Ti+JQ1 group had significantly lower mRNA expression levels(F=24.575-336.704,P<0.05).The Ti group had a significantly higher number of TRAP-positive osteoclasts than the other two groups(F=165.941,P<0.05).The Ti group had a significantly larger area of bone resorption than the control group,and the Ti+JQ1 group had a significantly smaller area of bone resorption than the Ti group(F=49.879,P<0.05).Conclusion Ti particles promote osteoclast formation and bone resorption by upregulating the expression of B

关 键 词：溴结构域蛋白4 钛颗粒 JQ1 破骨细胞 骨吸收 基因检测 

分 类 号：R687.4[医药卫生—骨科学] R318[医药卫生—外科学]