Introduction: Bone fracture is a common reason causing human disability. The delay union and nonunion rates are approximately 5– 10% despite patients receiving active treatment. Currently, there is a limited number of drugs directly accelerating bone healing, especially direct extracts from plants. Moreover, the pharmacological effects of Ilex cornuta bark are still unknown. This study aimed to explore the effects and mechanisms of Ilex cornuta bark in bone healing.
Methods and Results: First, the promoting effects of Ilex cornuta bark on bone healing were verified by the mice femur fracture model as Ilex cornuta bark increased the callus formation and enhanced the biomechanical stability during the bone healing process. Second, the target gene of Ilex cornuta bark in bone healing identified by bioinformatics analysis and immunofluorescence validation was ADORA2A. Third, 410 main compound compositions of Ilex cornuta bark were explored by a non-target metabolomic analysis, where 190 of them were neg ion mode, and 220 were pos ion mode. Molecular docking was used to predict the regulatory effect of the compounds on adora2a (adenosine A2A receptor), and ursonic acid had the lowest binding energy with adora2a. Finally, nfkb1 was the transcription factor (TF) of adora2a, and ursonic acid also had the lowest binding energy by bioinformatic analysis and molecular docking.
Conclusion: Overall, Ilex cornuta bark water extract was a new plant extract on promoting bone healing; in addition, the mechanism of it might be activating adora2a though Nfkb1.

Keywords: Ilex cornuta bark, adenosine A2A receptor, bone healing, non-target metabolomic analysis, bioinformatic analysis, Nfkb1


中文翻译：

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冬青树皮提取物通过激活腺苷 A2A 受体促进骨愈合

简介：骨折是导致人类残疾的常见原因。尽管患者接受了积极治疗，但延迟愈合和不愈合率约为 5-10%。目前，直接加速骨愈合的药物数量有限，尤其是植物的直接提取物。此外，冬青树皮的药理作用尚不清楚。本研究旨在探讨冬青树皮在骨愈合中的作用和机制。
方法与结果：首先，通过小鼠股骨骨折模型验证冬青树皮对骨愈合的促进作用。树皮在骨愈合过程中增加了愈伤组织的形成并增强了生物力学稳定性。其次，通过生物信息学分析和免疫荧光验证确定的冬青树皮在骨愈合中的靶基因是ADORA2A。第三，通过非目标代谢组学分析探索了410种冬青树皮的主要化合物组成，其中190种为负离子模式，220种为正离子模式。分子对接用于预测化合物对adora2a（腺苷A2A受体）的调节作用，熊果酸与adora2a的结合能最低。最后，通过生物信息学分析和分子对接，nfkb1是adora2a的转录因子（TF），熊果酸的结合能也最低。
结论：总体而言，冬青树皮水提取物是一种促进骨愈合的新型植物提取物；此外，它的机制可能是通过 Nfkb1 激活 adora2a。

关键词： 冬青树皮，腺苷A2A受体，骨愈合，非靶向代谢组学分析，生物信息学分析，Nfkb1