Hu Xueling, Lei Xiaotong, Lin Weiwen, Li Xiaoyun, Zhong Wenqiang, Luo Bingjie, Xie Ji, Liang Ziwen, Li Yunchuan, Qiu Jingli, Wang Panpan, Zhu Xiaofeng, Zhang Ronghua, Yang Li
College of Pharmacy, Jinan University, Guangzhou, 510632, China; Guangdong Provincial Key Laboratory of Traditional Chinese Medicine Informatization, Guangzhou, 510632, China.
Guangdong Provincial Key Laboratory of Traditional Chinese Medicine Informatization, Guangzhou, 510632, China; College of Traditional Chinese Medicine, Jinan University, Guangzhou, 510632, China.
Exp Cell Res. 2025 Jan 15;444(2):114386. doi: 10.1016/j.yexcr.2024.114386. Epub 2024 Dec 16.
Postmenopausal osteoporosis, primarily driven by estrogen deficiency, is predominantly mediated through estrogen receptors such as ERα. However, the underlying mechanisms necessitate further investigation. In this study, we established an ERα-deficient model in rBMSCs to elucidate the role of ERα in osteogenic differentiation and miRNA expression profiles. Our findings demonstrate that knockdown of ERα inhibits osteogenic differentiation in rBMSCs, resulting in upregulation of 25 miRNAs and downregulation of 184 miRNAs, including a significant increase in the expression of miR-214-3p. Validation using qPCR, Western blotting, and bioinformatics analysis revealed that miR-214-3p negatively regulates osteogenic differentiation via the Wnt/β-catenin signaling pathway. Furthermore, we explored the potential therapeutic effects of quercetin (QUE) on rBMSCs. CCK8, alkaline phosphatase activity assays, and Alizarin Red staining demonstrated that QUE dose-dependently enhances rBMSCs proliferation, alkaline phosphatase activity, and mineralization within the concentration range of 0.1-1 μM. Importantly, QUE was found to downregulate miR-214-3p expression and activate the Wnt3a/β-catenin signaling pathway. Rescue experiments confirmed that QUE could counteract the inhibitory effects of miR-214-3p on the Wnt3a/β-catenin signaling pathway. Collectively, our study provides compelling evidence that knockdown of ERα inhibits the osteogenic differentiation of rBMSCs by affecting the miRNA expression profile, while QUE can reverse the inhibitory effect exerted by miR-214-3p on the Wnt3a/β-catenin signaling pathway, thereby offering novel insights into diagnosis, prevention, and treatment strategies for postmenopausal osteoporosis.
绝经后骨质疏松症主要由雌激素缺乏引起，主要通过雌激素受体如ERα介导。然而，其潜在机制仍需进一步研究。在本研究中，我们在rBMSCs中建立了ERα缺陷模型，以阐明ERα在成骨分化和miRNA表达谱中的作用。我们的研究结果表明，敲低ERα会抑制rBMSCs的成骨分化，导致25种miRNA上调和184种miRNA下调，包括miR-214-3p表达显著增加。使用qPCR、蛋白质免疫印迹和生物信息学分析进行验证，结果显示miR-214-3p通过Wnt/β-连环蛋白信号通路负调控成骨分化。此外，我们探索了槲皮素（QUE）对rBMSCs的潜在治疗作用。CCK8、碱性磷酸酶活性测定和茜素红染色表明，在0.1-1μM浓度范围内，QUE剂量依赖性地增强rBMSCs的增殖、碱性磷酸酶活性和矿化。重要的是，发现QUE可下调miR-214-3p表达并激活Wnt3a/β-连环蛋白信号通路。挽救实验证实，QUE可以抵消miR-214-3p对Wnt3a/β-连环蛋白信号通路的抑制作用。总的来说，我们的研究提供了有力的证据，即敲低ERα通过影响miRNA表达谱抑制rBMSCs的成骨分化，而QUE可以逆转miR-214-3p对Wnt3a/β-连环蛋白信号通路的抑制作用，从而为绝经后骨质疏松症的诊断、预防和治疗策略提供了新见解。
