甘草苷抑制破骨细胞分化缓解骨丢失  

Liquiritin inhibits osteoclast differentiation and alleviates bone loss

作　　者：张文胜 郭海威 翁汭 莫凌 宋振杰[1,2,3,4] 田瀚 钟业霖[2,3,4] 王元成 唐汉武 刘才俊 原超 李颖[1,2,3,4] Zhang Wensheng;Guo Haiwei;Weng Rui;Mo Ling;Song Zhenjie;Tian Han;Zhong Yelin;Wang Yuancheng;Tang Hanwu;Liu Caijun;Yuan Chao;Li Ying(Guangdong Provincial Traditional Chinese Medicine Orthopedic Research Institute,Guangzhou 510240,Guangdong Province,China;Third Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510240,Guangdong Province,China;Guangzhou University of Chinese Medicine,Guangzhou 510006,Guangdong Province,China;Lingnan Medical Research Center,Guangzhou 510006,Guangdong Province,China)

机构地区：[1]广东省中医骨伤研究院,广东省广州市510240 [2]广州中医药大学第三附属医院,广州中医药大学第三临床医学院,广东省广州市510240 [3]广州中医药大学,广东省广州市510006 [4]岭南医学研究中心,广东省广州市510006

出　　处：《中国组织工程研究》2025年第12期2429-2437,共9页Chinese Journal of Tissue Engineering Research

基　　金：广东省自然科学基金面上项目(2022A1515011404),项目负责人:李颖;广东省中医骨伤研究院开放基金(GYH202102-04),项目负责人:原超;广东省中医骨伤研究院开放基金(GYH202101-03),项目负责人:李颖。

摘　　要：背景:破骨细胞的骨吸收功能相对或绝对活跃是骨质疏松症的诱因之一,因此,如何抑制破骨细胞形成、降低骨吸收活性是预防和治疗骨质疏松症的重点内容。甘草苷来源于传统中药甘草,对临床骨相关疾病具有治疗作用,但目前关于甘草苷在骨质疏松症方面的研究较少且机制不明。目的:通过体内外实验共同验证甘草苷抑制破骨细胞分化和缓解骨丢失的作用。方法:CCK-8实验检测甘草苷对小鼠来源骨髓巨噬单核细胞是否具有毒性或增殖作用,抗酒石酸酸性磷酸酶染色观察甘草苷抑制破骨细胞分化情况;通过网络药理学验证甘草苷与破骨细胞分化相关蛋白结合的亲和力大小;RT-PCR和Western blot实验检测甘草苷对破骨细胞相关特异性蛋白和基因表达以及相关信号通路的抑制作用;最后通过C57BL/6J小鼠骨质疏松模型验证甘草苷缓解骨丢失的作用。结果与结论:甘草苷在浓度20μmol/L及以下时能够起到抑制破骨细胞形成和分化的作用,同时与破骨细胞相关特异性蛋白如活化T细胞核因子c1、组织蛋白酶K、c-Fos、基质金属蛋白酶9具有较高的亲和力,能够降低上述基因和蛋白的相对表达水平,甘草苷也能够在干预第15,30,45,60分钟时有效降低MAPK信号通路中JNK的磷酸化表达水平,同时在第60分钟时能够挽救核因子κB信号通路中IκB-α的降解现象,体内实验证明甘草苷能够缓解由破骨细胞引起的骨质丢失现象,改善骨小梁相关参数。结果表明:甘草苷能抑制破骨细胞分化和缓解骨丢失,从而起到抗骨质疏松症的作用。BACKGROUND:Relatively or absolutely active bone resorption function of osteoclasts is one of the causative factors of osteoporosis.Therefore,how to inhibit the formation of osteoclasts and reduce the bone resorption activity is a key element in the prevention and treatment of osteoporosis.Liquiritin,which is derived from licorice,plays a role in the clinical treatment of bone diseases,but there are fewer studies addressing the application of liquiritin in osteoporosis and the mechanism is unknown.OBJECTIVE:To confirm,through both in vivo and in vitro experiments,that liquiritin inhibits osteoclast differentiation and alleviates bone loss.METHODS:Cell counting kit-8 was used to detect whether Liquiritin exerts toxic or proliferative effects on mouse bone marrow-derived macrophages,and tartrate-resistant acid phosphatase staining was performed to observe the effect of liquiritin in inhibiting osteoclast differentiation.The affinity of liquiritin binding to proteins related to osteoclast differentiation was verified by network pharmacology.RT-PCR and western blot assays were performed to detect the inhibitory effects of liquiritin on osteoclast-specific protein and gene expression as well as relevant signaling pathways.Finally,the mitigating effect of liquiritin on bone loss was verified in the C57BL/6J mouse osteoporosis model.RESULTS AND CONCLUSION:Liquiritin,at concentrations of 20μmol/L and below,could inhibit the formation and differentiation of osteoclasts.Concurrently,it exhibited a high affinity with osteoclast-specific proteins such as nuclear factor of activated T-cells 1,Cathepsin K,c-Fos,and matrix metalloproteinase 9,and reduced the relative expression levels of these genes and proteins.Liquiritin could also effectively lower the phosphorylation expression level of JNK in the MAPK signaling pathway at the 15th,30th,45th,and 60th minutes,and it could salvage the degradation of nuclear factor-κB inhibitorαin the nuclear factor-κB signaling pathway at the 60th minute.In vivo experiments demonstrated th

关 键 词：骨质疏松症 破骨细胞分化 甘草苷 骨重塑 网络药理学 分子对接 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R34