新补骨脂异黄酮对破骨细胞形成及破骨基因表达的作用  

Effects of Neobavaisoflavone on Osteoclast Formation and Osteoclast Gene Expression

作　　者：陈国材[1,2] CHEN Guo-cai(The Eighth Clinical Medical College of Guangzhou University of Chinese Medicine,Foshan,Guangdong 528000;The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou,Guangdong 510405)

机构地区：[1]广州中医药大学第八临床医学院,广东佛山528000 [2]广州中医药大学第一附属医院,广东广州510405

出　　处：《中医康复》2024年第5期5-9,15,共6页Traditional Chinese Medicine Rehabilitation

基　　金：广东省基础与应用基础研究基金(2021A1515110142)。

摘　　要：目的:探讨新补骨脂异黄酮对破骨细胞形成及破骨相关基因表达的影响。方法:采用CCK-8法检测不同浓度新补骨脂异黄酮对RAW 264.7细胞增殖的影响,并筛选出最佳干预浓度;通过TRAP活性染色和细胞骨架F-actin染色评估新补骨脂异黄酮对破骨细胞形成的影响;通过实时定量PCR技术检测破骨细胞分化相关基因基质金属蛋白酶9、组织蛋白酶K、和原癌基因c-Fos的mRNA表达。结果:在12.5μM及以下的浓度,新补骨脂异黄酮对RAW 264.7细胞无显著毒性;TRAP染色活性染色结果表明,新补骨脂异黄酮可以抑制破骨细胞分化;细胞骨架F-actin染色结果表明,新补骨脂异黄酮减小破骨细胞肌动蛋白纤维环的产生,抑制了破骨分化;新补骨脂异黄酮抑制原癌基因c-Fos(P<0.05),显著抑制基质金属蛋白酶9和组织蛋白酶K(P<0.01)等破骨细胞分化的mRNA的表达。结论:新补骨脂异黄酮能够在体外抑制破骨相关基因的表达,进而抑制破骨细胞的生成。Objective:To explore the effects of Neobavaisoflavone on osteoclast formation and expression of osteoclast-related genes.Methods:The CCK-8 method was used to detect the effect of different concentrations of Neobavaisoflavone on RAW 264.7 cell proliferation,and the optimal intervention concentration was selected.The effect of Neobavaisoflavone on osteoclast formation was evaluated by TRAP activity staining and cytoskeletal F-actin staining.Real-time quantitative PCR technology was used to detect the mRNA expression of osteoclast differentiation-related genes matrix metalloproteinase 9,cathepsin K,and proto-oncogene c-Fos.Results:At a concentration of 12.5μM and below,Neobavaisoflavone have no significant toxicity to RAW 264.7 cells.TRAP staining activity staining results showed that Neobavaisoflavone can inhibit osteoclast differentiation.Cytoskeletal F-actin staining results showed that Neobavaisoflavone reduced the production of actin fibrous annulus in osteoclasts and inhibited osteoclast differentiation.Neobavaisoflavone inhibits the proto-oncogene c-Fos(P<0.05)and significantly inhibits the expression of osteoclast differentiation mRNA such as matrix metalloproteinase 9 and cathepsin K(P<0.01).Conclusion:Neobavaisoflavone can inhibit the expression of osteoclast-related genes in vitro,thereby inhibiting the generation of osteoclasts.

关 键 词：新补骨脂异黄酮 破骨细胞分化 RAW 264.7细胞 骨质疏松症 

分 类 号：R285[医药卫生—中药学]