摘要
目的基于SIRT2-C/EBPβ-AREG轴探讨肉苁蓉苯乙醇苷(CPhGs)通过调控成骨分化改善骨质疏松的作用机制。方法采用卵巢切除法建立小鼠骨质疏松模型。分别采用micro-CT、钙黄绿素双标记和苏木素-伊红染色评估骨小梁数量/厚度、骨形成率和组织形态。采用Western blotting、免疫共沉淀检测SIRT2-C/EBPβ-AREG轴相关蛋白SIRT2、C/EBPβ、AREG表达。采用实时定量PCR检测成骨分化标志基因OCN、OPN、RUNX2、C/EBPβ、AREG mRNA表达。结果CPhGs干预显著改善卵巢切除小鼠骨小梁微结构并促进骨形成,其机制涉及激活SIRT2介导的C/EBPβ去乙酰化,进而上调AREG表达。细胞实验证实,CPhGs通过增强SIRT2与C/EBPβ的相互作用,显著提高MC3T3-E1细胞中碱性磷酸酶活性及OCN、OPN、RUNX2等成骨基因表达,且该效应可被SIRT2敲低逆转。结论CPhGs通过SIRT2-C/EBPβ-AREG轴调控成骨分化进程,为中药活性成分治疗骨质疏松症提供了新的分子靶点和理论依据。
Objective To explore the mechanism of Cistanche phenylethanoid glycosides(CPhGs)in treating osteoporosis by regulating osteogenic differentiation via SIRT2-C/EBPβ-AREG axis.Methods An osteoporosis mouse model was established using ovariectomy.Trabecular number/thickness,bone formation rate,and tissue morphology were evaluated using micro-computed tomography,calcein double labeling,and hematoxylin and eosin staining,respectively.SIRT2,C/EBPβ,AREG,proteins related to the SIRT2-C/EBPβ-AREG axis,were analyzed by Western blotting and co-immunoprecipitation.The mRNA expression of osteogenic differentiation marker genes OCN,OPN,RUNX2,C/EBPβ,and AREG were detected by real-time quantitative PCR.Results CPhGs intervention significantly improved the trabecular microarchitecture and promoted bone formation in mice undergoing ovariectomy,and the mechanism involved the activation of SIRT2-mediated deacetylation of C/EBPβ,which in turn upregulated AREG expression.Cell experiments confirmed that CPhGs significantly increased the activity of alkaline phosphatase and the expression of osteogenic genes such as OCN,OPN,and RUNX2 in MC3T3-E1 cells by enhancing the interaction between SIRT2 and C/EBPβ.Notably,this effect could be reversed by SIRT2 knockdown.Conclusion CPhGs regulate the osteogenic differentiation process through the SIRT2-C/EBPβ-AREG axis,providing a new molecular target and theoretical basis for the treatment of osteoporosis with the active ingredients of traditional Chinese medicine.
作者
章跃
叶里子
罗雷茗
王远辉
王治
李忠
ZHANG Yue;YE Lizi;LUO Leiming;WANG Yuanhui;WANG Zhi;LI Zhong(Department of Pediatric Surgery,Affiliated Hospital of Southwest Medical University,Luzhou 646000,China;Department of Bone and Joint Surgery,Affiliated Hospital of Southwest Medical University,Luzhou 646000,China)
出处
《中国医科大学学报》
北大核心
2025年第11期1011-1016,1022,共7页
Journal of China Medical University
基金
四川省省级科技计划(2023YFQ0009)。
