Xie An-Na, Zhang Sun-Zheng-Yuan, Zhang Yu, Cao Jin-Long, Wang Cheng-Long, Wang Li-Bo, Wu Hong-Jin, Zhang Jie, Dai Wei-Wei
Laboratory of Science and Technology Center, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China.
Laboratory of Science and Technology Center, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China.
J Integr Med. 2025 Aug 13. doi: 10.1016/j.joim.2025.08.002.
Glucocorticoid-induced osteoporosis (GIOP) is a common complication of prolonged glucocorticoid therapy. Chlorogenic acid (CGA), a polyphenol with antioxidant properties that is extracted from traditional Chinese medicines such as Eucommiae Cortex, has potential anti-osteoporotic activity. This study aimed to investigate the possible effects of CGA on GIOP in mice and murine long bone osteocyte Y4 (MLO-Y4) cells and explore the underlying molecular mechanisms.
The protective effects of CGA were initially evaluated in the GIOP mouse model induced by dexamethasone (Dex). The micro-computed tomography, hematoxylin-eosin staining, silver nitrate staining, and serum detection were used to assess the efficacy of CGA for improving bone formation in vivo. Then, network pharmacology analysis was used to predict the potential targets and molecular mechanisms underlying the therapeutic efficacy of CGA against GIOP. After that, 2',7'-dichlorofluorescein diacetate staining, flow cytometry, real-time quantitative reverse transcription polymerase chain reaction, and Western blotting were used to verify the mechanisms of CGA against GIOP in vitro.
Animal experiments showed that CGA treatment effectively attenuated Dex-induced decreases in bone mass and strength and improved disrupted osteocyte morphology in mice. The protein-protein interaction analysis highlighted erb-b2 receptor tyrosine kinase (ERBB2), which is also known as human epidermal growth factor receptor 2 (HER2), caspase-3, kinase insert domain receptor, matrix metallopeptidase 9, matrix metallopeptidase 2, proto-oncogene tyrosine-protein kinase Src, and epidermal growth factor receptor as core targets. The Kyoto Encyclopedia of Genes and Genomes analysis revealed several significantly enriched pathways (P < 0.05), including the ERBB, phosphoinositide 3 kinase-AKT serine/threonine kinase 1 (AKT), and mechanistic target of rapamycin kinase (mTOR) pathways. Cellular experiments verified that CGA enhanced bone formation and promoted autophagy while inhibiting apoptosis in MLO-Y4 cells exposed to Dex, which was associated with the upregulated expression of HER2 and activation of the HER2/AKT/mTOR signaling pathway.
CGA exerted anti-osteoporotic effects against GIOP, partially through targeting osteocytes and modulating the HER2/AKT/mTOR signaling pathway. Please cite this article as: Xie AN, Zhang SZY, Zhang Y, Cao JL, Wang CL, Wang LB, Wu HJ, Zhang J, Dai WW. Chlorogenic acid mitigates glucocorticoid-induced osteoporosis via modulation of HER2/AKT/mTOR signaling pathway. J Integr Med. 2025; Epub ahead of print.
糖皮质激素性骨质疏松症（GIOP）是长期糖皮质激素治疗常见的并发症。绿原酸（CGA）是一种具有抗氧化特性的多酚，从杜仲等中药中提取，具有潜在的抗骨质疏松活性。本研究旨在探讨CGA对小鼠GIOP及小鼠长骨骨细胞Y4（MLO-Y4）细胞的可能影响，并探索其潜在的分子机制。
首先在由地塞米松（Dex）诱导的GIOP小鼠模型中评估CGA的保护作用。采用显微计算机断层扫描、苏木精-伊红染色、硝酸银染色和血清检测来评估CGA在体内改善骨形成的效果。然后，运用网络药理学分析预测CGA治疗GIOP疗效的潜在靶点和分子机制。之后，采用2',7'-二氯荧光素二乙酸酯染色、流式细胞术、实时定量逆转录聚合酶链反应和蛋白质印迹法在体外验证CGA抗GIOP的机制。
动物实验表明，CGA治疗有效减轻了Dex诱导的小鼠骨量和骨强度下降，并改善了骨细胞形态破坏。蛋白质-蛋白质相互作用分析突出了erb-b2受体酪氨酸激酶（ERBB2，也称为人表皮生长因子受体2（HER2））、半胱天冬酶-3、激酶插入结构域受体、基质金属肽酶9、基质金属肽酶2、原癌基因酪氨酸蛋白激酶Src和表皮生长因子受体作为核心靶点。京都基因与基因组百科全书分析揭示了几个显著富集的通路（P < 0.05），包括ERBB、磷脂酰肌醇3激酶-AKT丝氨酸/苏氨酸激酶1（AKT）和雷帕霉素靶蛋白激酶（mTOR）通路。细胞实验证实，CGA增强了暴露于Dex的MLO-Y4细胞的骨形成并促进自噬，同时抑制细胞凋亡，这与HER2表达上调和HER2/AKT/mTOR信号通路激活有关。
CGA对GIOP具有抗骨质疏松作用，部分是通过靶向骨细胞并调节HER2/AKT/mTOR信号通路实现的。请引用本文：Xie AN, Zhang SZY, Zhang Y, Cao JL, Wang CL, Wang LB, Wu HJ, Zhang J, Dai WW. Chlorogenic acid mitigates glucocorticoid-induced osteoporosis via modulation of HER2/AKT/mTOR signaling pathway. J Integr Med. 2025; Epub ahead of print.
