Shi Jingcun, Wen Jin, Hu Longwei
Department of Oral and Maxillofacial Surgery - Head & Neck Oncology, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, 639 Zhizaoju Road, Shanghai, 200011, China.
College of Stomatology, National Center for Stomatology, Shanghai Jiao Tong University, National Clinical Research Center for Oral Diseases, Shanghai, China.
J Orthop Surg Res. 2025 Jan 10;20(1):35. doi: 10.1186/s13018-024-05400-9.
The study aims to elucidate the mechanism through which 17β-estradiol facilitates osteogenic differentiation in bone marrow mesenchymal stem cells (BMSCs). In our study, lentiviral transfection was employed to establish apoptosis repressor with caspase recruitment domain (ARC) knockdown or overexpression in BMSCs. The impact of 17β-estradiol on ARC expression was assessed using western blot, RT-PCR and immunofluorescence. Techniques such as ALP staining, ALP activity assay, western blot, RT-PCR and immunofluorescence staining were utilized to examine the influence of ARC expression levels on the osteogenic differentiation of BMSCs and the osteoclastic differentiation of Raw264.7 cell lines. Mitophagy flux levels in BMSCs were detected using the mitophagy detection kit. RNA sequencing and bioinformatics analyses were conducted to explore potential mechanisms of ARC regulation in BMSCs osteogenic differentiation. To sum up, 17β-estradiol can modulate bone homeostasis by adjusting ARC expression. ARC stimulates mitophagy in BMSCs via MAPK/Akt pathway, identifying ARC as a promising therapeutic target for postmenopausal osteoporosis (PMOP) treatment.
该研究旨在阐明17β-雌二醇促进骨髓间充质干细胞（BMSCs）成骨分化的机制。在我们的研究中，采用慢病毒转染在BMSCs中建立具有半胱天冬酶募集结构域（ARC）的凋亡抑制因子敲低或过表达。使用蛋白质免疫印迹法、逆转录-聚合酶链反应（RT-PCR）和免疫荧光评估17β-雌二醇对ARC表达的影响。利用碱性磷酸酶（ALP）染色、ALP活性测定、蛋白质免疫印迹法、RT-PCR和免疫荧光染色等技术检测ARC表达水平对BMSCs成骨分化及Raw264.7细胞系破骨细胞分化的影响。使用线粒体自噬检测试剂盒检测BMSCs中的线粒体自噬通量水平。进行RNA测序和生物信息学分析以探索ARC调控BMSCs成骨分化的潜在机制。综上所述，17β-雌二醇可通过调节ARC表达来调节骨稳态。ARC通过丝裂原活化蛋白激酶/蛋白激酶B（MAPK/Akt）途径刺激BMSCs中的线粒体自噬，确定ARC为绝经后骨质疏松症（PMOP）治疗的一个有前景的治疗靶点。
