淫羊藿苷通过caveolin-1/Hippo信号通路促进BMSCs成骨分化改善骨代谢紊乱的机制研究  

Mechanism of icariin in promoting osteogenic differentiation of BMSCs and improving bone metabolism disorders through caveolin-1/Hippo signaling pathway

作　　者：韩一旦 张海凤 许云腾 钟玉环 王晓宁[1] 俞允[1] 严媛丽 王珊珊 李西海[2,3] HAN Yi-dan;ZHANG Hai-feng;XU Yun-teng;ZHONG Yu-huan;WANG Xiao-ning;YU Yun;YAN Yuan-li;WANG Shan-shan;LI Xi-hai(College of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Science and Technology Department,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Fujian Key Laboratory of Integrative Medicine on Geriatrics,Fuzhou 350122,China)

机构地区：[1]福建中医药大学中西医结合学院,福建福州350122 [2]福建中医药大学科学技术处,福建福州350122 [3]福建省中西医结合老年性疾病重点实验室,福建福州350122

出　　处：《中国中药杂志》2025年第3期600-608,共9页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82304899);福建省自然科学基金面上项目(2021J01893);福建中医药大学高层次人才引进项目(X2020011-人才)。

摘　　要：该实验以“肾藏精主骨生髓”为指导,通过体内体外实验,探究淫羊藿苷(icariin,ICA)通过小窝蛋白(caveolin-1,Cav1)调控大鼠骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)成骨分化机制,旨在为中医药防治绝经后骨质疏松症提供理论依据。采用全骨髓贴壁法获取4周龄SD雌性大鼠原代细胞。流式细胞术检测表面标志物CD29、CD90、CD11b、CD45表达;成骨成脂分化检测分化潜能。CCK-8检测ICA对细胞活性影响;茜素红染色验证ICA对矿化结节形成影响。慢病毒沉默Cav1构建稳转细胞株。茜素红染色观察沉默Cav1对成骨分化影响。Western blot检测Cav1、Hippo/TAZ、成骨指标Runt相关转录因子2(runt-related transcription factor 2,RUNX2)、碱性磷酸酶(alkaline phosphatase,ALP)蛋白表达。结果显示,通过全骨髓贴壁法成功获取原代细胞,阳性表达大鼠BMSCs表面标志物,具有成骨成脂多向分化潜能。CCK-8结果和茜素红染色结果显示1×10^(-7)mol·L^(-1)为ICA最佳干预浓度(P<0.05)。成骨诱导分化过程中,ICA抑制Cav1表达(P<0.05),促进TAZ表达(P<0.05)。茜素红染色证明沉默Cav1明显促进BMSCs成骨分化;ICA干预后TAZ表达激活,成骨指标ALP、RUNX2表达升高。综上所述,沉默Cav1显著促进BMSCs成骨分化,ICA通过抑制Cav1调控Hippo/TAZ信号通路促进BMSCs成骨分化。Guided by the theory of "the kidney storing essence, governing the bones, and producing marrow", this study explored the mechanism of icariin(ICA) in regulating the osteogenic differentiation of rat bone mesenchymal stem cells(BMSCs) through caveolin-1(Cav1) via in vitro and in vivo experiments, aiming to provide a theoretical basis for the prevention and treatment of postmenopausal osteoporosis with traditional Chinese medicine(TCM). Primary cells were obtained from 4-week-old female SD rats using the whole bone marrow adherent method. Flow cytometry was used to detect the expression of surface markers CD29, CD90, CD11b, and CD45. The potential for osteogenic and adipogenic differentiation was assessed. The effect of ICA on cell viability was determined using the CCK-8 assay, and the impact of ICA on the formation of mineralized nodules was verified by alizarin red staining. A stable Cav1-silenced cell line was constructed using lentivirus. The effect of Cav1 silencing on osteogenic differentiation was observed via alizarin red staining. Western blot analysis was conducted to detect the expression of Cav1, Hippo/TAZ, and osteogenic markers such as Runt-related transcription factor 2(RUNX2) and alkaline phosphatase(ALP). The results showed that primary cells were successfully obtained using the whole bone marrow adherent method, positively expressing surface markers of rat BMSCs and possessing the potential for both osteogenic and adipogenic differentiation. The CCK-8 assay and alizarin red staining results indicated that 1×10^(-7) mol·L^(-1) was the optimal concentration of ICA for intervention in this experiment(P<0.05). During osteogenic induction, ICA inhibited Cav1 expression(P<0.05) while promoting TAZ expression(P<0.05). Alizarin red staining demonstrated that Cav1 silencing significantly promoted the osteogenic differentiation of BMSCs. After ICA intervention, TAZ expression was activated, and the expression of osteogenic markers ALP and RUNX2 was increased. In conclusion, Cav1 silencing significantly p

关 键 词：绝经后骨质疏松症 淫羊藿苷 骨髓间充质干细胞 成骨分化 CAVEOLIN-1 Hippo/TAZ 

分 类 号：R285.5[医药卫生—中药学]