基于骨免疫的抗骨质疏松活性天然产物的筛选及功能验证

Screening and functional validation of active natural products against osteoporosis based on osteoimmunology

摘要

目的 基于系统药理学筛选作用于骨质疏松症骨免疫基因的活性天然产物, 并研究其抗骨质疏松的功能。方法 基于GEO数据库骨质疏松症患者芯片数据集和GeneCards数据库获得骨免疫基因。采用SysDT模型筛选作用于骨免疫基因的活性天然产物, 再结合口服生物利用度≥30%和类药性≥0.18, 筛选潜在的抗骨质疏松活性天然产物。进一步, 采用前成骨细胞系MC3T3-E1细胞, 研究所筛选的活性天然产物杜鹃素和吴茱萸次碱对成骨细胞分化的影响。以大鼠后肢去负荷构建大鼠废用模型, 探究活性天然产物杜鹃素对废用性骨质疏松的防治效果。结果 骨质疏松症患者的芯片数据分析获得272个骨免疫基因, 进一步筛选出作用于骨免疫基因的潜在活性天然产物杜鹃素和吴茱萸次碱。体外实验发现杜鹃素和吴茱萸次碱均能促进成骨细胞碱性磷酸酶的表达和成骨分化基因的表达(P<0.05, P<0.01); 表明其均能促进成骨细胞分化, 并且杜鹃素优于吴茱萸次碱。进一步的体内实验表明杜鹃素能显著增强尾悬吊大鼠骨密度和骨矿物质含量(P<0.05), 改善骨生物力学性能(P<0.01), 并调控骨免疫因子肿瘤坏死因子α、白介素4、白介素10的表达(P<0.01)。结论 基于骨免疫基因筛选出的活性天然产物杜鹃素通过促进成骨细胞分化对抗骨质疏松症, 其可能为骨质疏松症的新药研发提供新思路。

Abstract

Objective To screen active natural products targeting osteoimmunology-related genes in osteoporosis using systems pharmacology, and to investigate their anti-osteoporotic functions. Methods Osteoimmunology-related genes were obtained from osteoporosis patient microarray datasets in the GEO database and the GeneCards database. Active natural products targeting these osteoimmunology-related genes were screened using the SysDT model. Potential anti-osteoporotic active natural products were further screened according to oral bioavailability≥30% and drug-likeness≥0.18. Furthermore, the pre-osteoblastic cell line MC3T3-E1 was used to investigate the effects of the screened active natural products, farrerol and rutaecarpine, on osteoblast differentiation. A rat model of disuse osteoporosis was established using the hindlimb unloading, and the preventive and therapeutic effects of the active natural product farrerol on disuse osteoporosis were explored. Results Microarray data analysis of osteoporosis patients identified 272 osteoimmunology-related genes. Further screening revealed two potential active natural products, farrerol and rutaecarpine, which target osteoimmunology-related genes. In vitro experiments revealed that both farrerol and rutaecarpine upregulated the expression of alkaline phosphatase and osteogenic differentiation-related genes in osteoblasts (P<0.05, P<0.01). These findings indicated that both farrerol and rutaecarpine facilitated osteoblast differentiation, with farrerol exhibiting a superior effect compared to rutaecarpine. Further in vivo experiments demonstrated that farrerol significantly increased bone mineral density and bone mineral content (P<0.05), improved bone biomechanical properties (P<0.01), and regulated the expression of bone immune factors including tumor necrosis factor-α, interleukin-4 and interleukin-10 in tail-suspended rats. Conclusion The active natural product farrerol, screened based on osteoimmunology-related genes, exerts anti-osteoporotic effects by promoting osteoblast differentiation, which may provide novel insights for the research and development of new drugs against osteoporosis.

关键词

骨免疫 / ADME筛选 / 抗骨质疏松 / 活性天然产物 / 系统药理学 / 后肢去负荷 / 杜鹃素 / 成骨细胞分化

Key words

osteoimmunology / ADME screening / anti-osteoporosis / active natural products / systems pharmacology / hindlimb unloading / farrerol / osteoblast differentiation

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基金资助

国家自然科学基金面上项目(82572148)

广东省基础与应用基础研究基金(2025A1515012343)