基于OPG/RANKL/RANK信号通路探讨益肾健骨方 防治 绝经后骨质疏松的作用机制  

Study of the mechanism of action of the tonifying kidney and strong bone formula in the prevention and treatment of postmenopausal osteoporosis based on OPG/RANKL/RANK signaling pathway

作　　者：叶子丰 戎宽[2] 张信成[2] 匡建军[2] 万明明 许辉[2] YE Zifeng;RONG Kuan;ZHANG Xincheng;KUANG Jianjun;WAN Mingming;XU Hui(Hunan University of Traditional Chinese Medicine,Changsha 410208;Yueyang Hospital of Traditional Chinese Medicine,Yueyang 410006,Hunan;The Affiliated Hospital of Hunan Institute of Traditional Chinese Medicine,Changsha 410006,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]湖南省中医药研究院附属医院,湖南长沙410006 [3]湖南省岳阳市中医医院,湖南岳阳414000

出　　处：《中国骨质疏松杂志》2023年第1期23-28,57,共7页Chinese Journal of Osteoporosis

基　　金：国家中医药管理局“全国名老中医药专家仇湘中传承工作室建设项目”(国中医药人教【2016】42号);湖南省西学中骨干人才项目资助(湘中医药函【2020】40号)。

摘　　要：目的通过研究益肾健骨方对破骨细胞的影响,进一步揭示益肾健骨方对绝经后骨质疏松症的作用机制。方法大鼠去双侧卵巢造模后分为实验组(去势+益肾健骨方灌胃)、模型组(去势+生理盐水灌胃)、假手术组(假手术+生理盐水灌胃)、阳性药物对照组(去势+阿仑膦酸钠灌胃),提取各组BMSCs进行培养、传代后备用,同时分离刚出生不久的大鼠破骨前体细胞进行体外培养,将各组骨髓间充质干细胞与破骨前体细胞共培养构建共培养体系。观察破骨细胞分化与成熟情况,并采用Real-time PCR与Western blot检测骨髓间充质干细胞与破骨前体细胞共培养体系中Wnt10b、β-Catenin、OPG、RANKL的表达情况。结果在共培养体系中模型+OC组Wnt10b、β-Catenin、OPG mRNA与蛋白表达最低,RANKL mRNA与蛋白表达最高,假手术+OC组表达最高,益肾健骨方与阳性对照物组处理后可提升Wnt10b、β-Catenin、OPG mRNA与蛋白的表达情况,且两组间比较差异无统计学意义。在共培养体系中模型+OC组RANKL mRNA与蛋白表达最高,假手术+OC组表达最低,益肾健骨方与阳性对照物组处理后可降低RANKL mRNA及蛋白的表达,且两组间比较差异无统计学意义。结论益肾健骨方可通过调节BMSCs抑制破骨细胞的形成,提高骨质量,同时通过激活OPG/RANKL/RANK信号通路抑制破骨细胞的分化与成熟,影响骨代谢从而防治PMOP。Objective To further reveal the mechanism of the tonifying kidney and strong bone formula in the prevention and treatment of postmenopausal osteoporosis by studying its effect on osteoclasts.Methods The rats were divided into the experimental group(ovariectomized+the tonifying kidney and strong bone formula by gavage),model group(ovariectomized+saline by gavage),sham-operated group(sham-operated+saline by gavage),and positive drug control group(ovariectomized+alendronate by gavage).BMSCs from each group were extracted for culture and passaged for use.BMSCs were co-cultured with osteoblastic precursor cells to build a co-culture system.The differentiation and maturation of osteoblasts were observed.The expressions of Wnt10b,β-catenin,OPG,and RANKL in the co-culture system of BMSCs and osteoblastic precursor cells was detected using Real-time PCR and Western blotting.Results In the co-culture system mRNA and protein expressions of Wnt10b,β-catenin,and OPG were the lowest in the model+OC group and the highest in the sham-operated+OC group.The treatment of the tonifying kidney and strong bone formula and alendronate in the positive control substance group enhanced the expressions of Wnt10b,β-catenin,and OPG mRNA and protein,but the difference between the two groups was not statistically significant.In the co-culture system,RANKL mRNA and protein expression were the highest in the model+OC group and the lowest in the sham operation+OC group.The treatment of the tonifying kidney and strong bone formula and alendronate in the positive control substance group reduced the expressions of RANKL mRNA and protein,but the difference between the two groups was not statistically significant.Conclusion By regulating the formation of osteoclasts and improving bone quality through BMSCs,the tonifying kidney and strong bone formula inhibits the differentiation and maturation of osteoclasts by activating the OPG/RANKL/RANK signaling pathway and by affecting bone metabolism,and thus preventing PMOP.

关 键 词：益肾健骨方 绝经后骨质疏松症 OPG/RANKL/RANK Wnt/β-catenin 作用机制 信号通路 

分 类 号：R285.5[医药卫生—中药学]