文献类型：期刊文献

中文题名：异甘草素抑制磨损颗粒诱导体内外破骨细胞的形成与分化

英文题名：Isoliquiritigenin inhibits wear particle-induced formation and differentiation of osteoblasts in vitro and in vivo

作者：谭飞[1,2];曾健康[1,2];王静[1,2];刘健[1,2];李嘉欢[1,2];李培杰[1,2];乔永杰[1];周胜虎[1]

第一作者：谭飞

机构：[1]解放军联勤保障部队第九四〇医院关节外科,甘肃省兰州市730050;[2]甘肃中医药大学第一临床医学院,甘肃省兰州市730030

第一机构：解放军联勤保障部队第九四〇医院关节外科,甘肃省兰州市730050

年份：2026

卷号：30

期号：11

起止页码：2691

中文期刊名：中国组织工程研究

外文期刊名：Chinese Journal of Tissue Engineering Research

收录：;北大核心:【北大核心2023】；

基金：甘肃省重点研发计划项目(25YFFA064),项目负责人:周胜虎;甘肃省卫健委项目(GSWSKY2024-20),项目负责人:周胜虎;联勤保障部队第九四〇医院高层次人才培养工程项目(2024-G3-5),项目负责人:乔永杰;兰州市科技计划项目(2023-ZD-173),项目负责人:乔永杰。

语种：中文

中文关键词：异甘草素;磨损颗粒;RAW264.7细胞;气囊植骨模型;无菌性松动

外文关键词：isoliquiritigenin;wear particle;RAW264.7 cell;air bag implant model;aseptic loosening

摘要：背景:磨损颗粒引起破骨细胞分化是导致假体周围骨溶解的主要原因之一,抑制破骨细胞的分化及吸收功能是治疗假体周围骨溶解的重要研究方向。研究显示,异甘草素可促进成骨细胞的成熟、抑制破骨细胞的骨吸收作用。目的:探讨异甘草素对磨损颗粒诱导破骨细胞形成与分化的影响。方法:①细胞实验:将第3代RAW264.7细胞分5组处理:对照组不进行任何处理,钛颗粒组加入钛颗粒,破骨诱导组加入核因子κB受体活化因子配体诱导破骨细胞分化,破骨诱导+钛颗粒组同时加入核因子κB受体活化因子配体与钛颗粒,异甘草素组加入核因子κB受体活化因子配体与钛颗粒处理第2天加入20μmol/L异甘草素。核因子κB受体活化因子配体诱导处理5 d后,抗酒石酸酸性磷酸酶、F-actin染色评估破骨细胞形成与骨吸收功能,Western Blot检测异甘草素对核因子κB通路中IκBα、p-65磷酸化的影响,ELISA法检测细胞上清液中炎症因子(肿瘤坏死因子α、白细胞介素6和基质金属蛋白酶9)水平,RT-PCR技术检测破骨细胞相关基因(组织蛋白酶K、活化T细胞核因子1、原癌基因蛋白及抗酒石酸酸性磷酸酶)表达。②动物实验:取32只BALB/c小鼠构建背部气囊植骨模型,随机分4组干预:空白组(n=8)气囊内注射PBS,模型组(n=8)、溶剂组(n=8)、异甘草素组(n=8)气囊内注射钛颗粒悬浮液,钛颗粒悬浮液注射第2天,空白组、模型组、溶剂组和异甘草素组分别腹腔注射PBS、PBS、DMSO+玉米油、40 mg/kg异甘草素,隔天注射1次,连续注射2周。干预结束后,抗酒石酸酸性磷酸酶染色检测气囊中骨片破骨细胞数量,ELISA法检测气囊中骨片研磨液中炎症因子水平,RT-PCR检测气囊中骨片研磨液中破骨细胞相关基因表达。结论与结果:①细胞实验:与对照组相比,其他4组破骨细胞形成数量增加、骨吸收能力增强,其中以破骨诱导+钛颗粒组破骨细胞形成数量最多、骨吸收能力最强;与破骨诱导+钛颗粒组相比,异甘草素组破骨细胞形成数量减少、骨吸收能力降低。与对照组相比,其余4组IκBα、p65磷酸化增加,提示破骨细胞数量增加,其中破骨诱导+钛颗粒组IκBα、p65磷酸化表达最多,异甘草素处理后明显降低。与对照组相比,其他4组炎症因子水平与破骨细胞相关基因表达均升高,其中以破骨诱导+钛颗粒组最高;与破骨诱导+钛颗粒组相比,异甘草素组炎症因子水平与破骨细胞相关基因表达均降低。②动物实验:与空白组相比,其他3组破骨细胞数量、炎症因子水平与破骨细胞相关基因表达均增加;与模型组相比,异甘草素组破骨细胞数量、炎症因子水平与破骨细胞相关基因表达均减少。③结果表明,异甘草素能够抑制磨损颗粒引发的体内外破骨细胞的形成与分化。
BACKGROUND:The differentiation of osteoclasts caused by wear particles is one of the main causes of periprosthesis osteolysis.Inhibiting the differentiation and absorption of osteoclasts is an important research direction in the treatment of periprosthesis osteolysis.Studies have shown that isoliquiritigenin can promote the maturation of osteoblasts and inhibit the bone resorption of osteoclasts.OBJECTIVE:To investigate the effect of isoliquiritigenin on osteoclast formation and differentiation induced by wear particles.METHODS:(1)Cell experiment:Passage 3 RAW264.7 cells were divided into five different groups.The control group was not treated with any treatment.The titanium particle group was added with titanium particles.The osteoclast induction group was added with nuclear factorκB receptor activating factor ligand to induce osteoclast differentiation.The osteoclast induction+titanium particle group was added with nuclear factorκB receptor activating factor ligand and titanium particles at the same time,and the isoliquiritigenin group was added with nuclear factorκB receptor activating factor ligand and titanium particles.On the 2nd day of treatment,20μmol/L isoliquiritigenin was added.After 5 days of induction with receptor activator of nuclear factor-κB ligand,tartrate-resistant acid phosphatase and F-actin staining were used to evaluate osteoclast formation and bone resorption.Western blot assay was used to detect the effect of isoliquiritigenin on the phosphorylation of IκBαand p-65 in the nuclear factor-κB pathway.Enzyme-linked immunosorbent assay was used to detect the levels of inflammatory factors(tumor necrosis factor-α,interleukin-6,and matrix metalloproteinase-9)in the cell supernatant.Reverse transcription polymerase chain reaction was used to detect the expressions of osteoclast-related genes(cathepsin K,nuclear factor of activated T cells 1,proto-oncogene protein,and tartrateresistant acid phosphatase).(2)Animal experiment:Totally 32 BALB/c mice were used to construct dorsal airbag bone graft models and randomly divided into four intervention groups:the blank group(n=8)was injected with PBS into the airbag,and the model group(n=8),solvent group(n=8),and isoliquiritigenin group(n=8)were injected with titanium particle suspension into the airbag.On the second day after the titanium particle suspension was injected,the blank group,model group,solvent group,and isoliquiritigenin group were intraperitoneally injected with phosphate buffered saline,phosphate buffered saline,dimethyl sulfoxide+corn oil,and 40 mg/kg isoliquiritigenin,respectively,once every other day for 2 consecutive weeks.After the intervention,tartrate-resistant acid phosphatase staining was used to detect the number of osteoclasts in the airbag.Enzyme-linked immunosorbent assay was used to detect the level of inflammatory factors in the grinding fluid of the airbag.Reverse transcription polymerase chain reaction was used to detect the expression of osteoclast-related genes in the grinding fluid of the airbag.RESULTS AND CONCLUSION:(1)Cell experiment:Compared with the control group,the number of osteoclasts formed in the other four groups increased,and the bone resorption capacity was enhanced.Among them,the osteoclast induction+titanium particle group had the largest number of osteoclasts formed and the strongest bone resorption capacity.Compared with the osteoclast induction+titanium particle group,the number of osteoclasts formed in the isoliquiritigenin group decreased,and the bone resorption capacity decreased.Compared with the control group,the phosphorylation of IκBαand p65 in the other four groups increased,indicating that the number of osteoclasts increased.Among them,the phosphorylation of IκBαand p65 in the osteoclast induction+titanium particle group was the highest,and it was significantly reduced after isoliquiritigenin treatment.Compared with the control group,the levels of inflammatory factors and the expression of osteoclast-related genes in the other four groups increased,among which the osteoclast induction+titanium particle group was the highest.Compared with the osteoclast induction+titanium particle group,the levels of inflammatory factors and the expression of osteoclast-related genes were reduced in the isoliquiritigenin group.(2)Animal experiment:Compared with the blank group,the number of osteoclasts,the level of inflammatory factors and the expression of osteoclast-related genes increased in the other three groups.Compared with the model group,the number of osteoclasts,the level of inflammatory factors and the expression of osteoclast-related genes decreased in the isoliquiritigenin group.These findings indicate that isoliquiritigenin could suppress the formation and differentiation of osteoclasts in vivo and in vitro induced by wear particles.