Jilin Medical Products Administration, Changchun 130000, China.
School of Pharmaceutical Sciences, Quality Evaluation & Standardization Hebei Province Engineering Research Center of Traditional Chinese Medicine, Hebei University of Chinese Medicine, Shijiazhuang 050091, China.
Molecules. 2023 Oct 9;28(19):6987. doi: 10.3390/molecules28196987.
Osteoporosis (OP) is distinguished by a reduction in bone mass and degradation of bone micro-structure, frequently resulting in fractures. As the geriatric demographic expands, the incidence of affected individuals progressively rises, thereby exerting a significant impact on the quality of life experienced by individuals. The flavonoid compound hesperidin has been subject to investigation regarding its effects on skeletal health, albeit the precise mechanisms through which it operates remain ambiguous. This study utilized network pharmacology to predict the core targets and signaling pathways implicated in the anti-OP properties of hesperidin. Molecular docking and molecular dynamics simulations were employed to confirm the stability of the interaction between hesperidin and the core targets. The effects of hesperidin on osteoblastic cells MC3T3-E1 were assessed using MTT, ELISA, alkaline phosphatase assay, and RT-qPCR techniques. Furthermore, in vivo experiments were conducted to determine the potential protective effects of hesperidin on zebrafish bone formation and oxidative stress response. The results demonstrate that network pharmacology has identified 10 key target points, significantly enriched in the estrogen signaling pathway. Hesperidin exhibits notable promotion of MC3T3-E1 cell proliferation and significantly enhances ALP activity. ELISA measurements indicate an elevation in NO levels and a reduction in IL-6 and TNF-α. Moreover, RT-qPCR analysis consistently reveals that hesperidin significantly modulates the mRNA levels of ESR1, SRC, AKT1, and NOS3 in MC3T3-E1 cells. Hesperidin promotes osteogenesis and reduces oxidative stress in zebrafish. Additionally, we validate the stable and tight binding of hesperidin with ESR1, SRC, AKT1, and NOS3 through molecular dynamics simulations. In conclusion, our comprehensive analysis provides evidence that hesperidin may exert its effects on alleviating OP through the activation of the estrogen signaling pathway via ESR1. This activation leads to the upregulation of SRC, AKT, and eNOS, resulting in an increase in NO levels. Furthermore, hesperidin promotes osteoblast-mediated bone formation and inhibits pro-inflammatory cytokines, thereby alleviating oxidative stress associated with OP.
骨质疏松症（OP）的特征是骨量减少和骨微结构降解，常导致骨折。随着老年人口的增加，受影响个体的发病率逐渐上升，从而对个体的生活质量产生重大影响。类黄酮化合物橙皮苷已被研究其对骨骼健康的影响，尽管其作用的确切机制仍不清楚。本研究利用网络药理学预测橙皮苷抗 OP 特性的核心靶点和信号通路。采用分子对接和分子动力学模拟验证橙皮苷与核心靶点相互作用的稳定性。采用 MTT、ELISA、碱性磷酸酶测定和 RT-qPCR 技术评估橙皮苷对成骨细胞 MC3T3-E1 的影响。此外，还进行了体内实验以确定橙皮苷对斑马鱼骨形成和氧化应激反应的潜在保护作用。结果表明，网络药理学已鉴定出 10 个关键靶标，这些靶标显著富集在雌激素信号通路中。橙皮苷显著促进 MC3T3-E1 细胞增殖，显著增强 ALP 活性。ELISA 测量表明 NO 水平升高，IL-6 和 TNF-α水平降低。此外，RT-qPCR 分析一致表明，橙皮苷显著调节 MC3T3-E1 细胞中 ESR1、SRC、AKT1 和 NOS3 的 mRNA 水平。橙皮苷促进斑马鱼成骨和减轻氧化应激。此外，我们通过分子动力学模拟验证了橙皮苷与 ESR1、SRC、AKT1 和 NOS3 的稳定和紧密结合。总之，我们的综合分析提供了证据表明，橙皮苷可能通过激活雌激素信号通路通过 ESR1 发挥其缓解 OP 的作用。这种激活导致 SRC、AKT 和 eNOS 的上调，从而增加 NO 水平。此外，橙皮苷促进成骨细胞介导的骨形成并抑制促炎细胞因子，从而减轻与 OP 相关的氧化应激。
