Farnesol Targets the GSTP1/MAPK Axis to Inhibit Trauma-Induced Tendon Heterotopic Ossification

Background: Heterotopic ossification (HO), characterized by pathological bone formation in extra-skeletal tissues such as tendon, often results in debilitating tissue dysfunction. HO of tendons primarily arises from abnormal osteogenic differentiation of tendon stem cells following injury. Despite being a common clinical occurrence, the cellular and molecular mechanisms of tendon HO are poorly understood, and effective treatments remain elusive. Farnesol, a natural isoprenoid alcohol abundant in citrus fruits, lavender, and royal jelly, is known for its potent anti-inflammatory, antioxidant, and antitumor properties. However, its applications in tissue engineering are rarely explored, particularly in musculoskeletal repair. Here, we provide the first report on the therapeutic potential of Farnesol in attenuating tendon HO and elucidate the associated molecular mechanisms.Methods: This study evaluated the effects of Farnesol on osteogenic differentiation of tendon-derived stem cells using both in vitro and in vivo models. The molecular mechanisms underlying the therapeutic effects of Farnesol on tendon HO were revealed by RNA sequencing combined with network pharmacology analysis, identifying the GSTP1/MAPK axis as a potential target pathway. In vitro rescue experiments using a specific GSTP1 inhibitor were conducted to confirm whether Farnesol primarily exerted its therapeutic effects through GSTP1.Results: The results demonstrated significant effects of Farnesol on attenuating tendon HO both in vitro and in vivo. Combined analysis of transcriptomics and network pharmacology indicated that Farnesol functions by targeting the GSTP1/MAPK signaling axis. These findings were further validated by in vitro experiments using the GSTP1 inhibitor, which reversed the suppressive effects of Farnesol on osteogenesis.Conclusion: Farnesol inhibits the progression of post-traumatic tendon HO by targeting the GSTP1/MAPK pathway. These findings provide new insights into the potential of using Farnesol as a therapeutic agent for HO. Overall design: RNA sequencing analysis was performed on the control group and farnesol-treated group of rat TDSCs on the third day of osteogenic differentiation.背景：异位骨化（Heterotopic ossification, HO）以肌腱等骨外组织内的病理性骨形成为特征，常导致致残性组织功能障碍。肌腱异位骨化主要源于损伤后肌腱干细胞的异常成骨分化。尽管该病症在临床中较为常见，但肌腱异位骨化的细胞与分子机制仍不甚明确，且尚无有效的治疗手段。法尼醇（Farnesol）是一种天然类异戊二烯醇，广泛存在于柑橘类水果、薰衣草及蜂王浆中，具有强效抗炎、抗氧化及抗肿瘤活性。但其在组织工程领域的应用鲜有探索，尤其在肌肉骨骼修复方向。本研究首次报道了法尼醇缓解肌腱异位骨化的治疗潜力，并阐明其相关分子机制。 方法：本研究通过体外与体内模型，评估法尼醇对肌腱源性干细胞（tendon-derived stem cells, TDSCs）成骨分化的影响。通过RNA测序（RNA sequencing）联合网络药理学（network pharmacology）分析，揭示了法尼醇治疗肌腱异位骨化的分子机制，确定谷胱甘肽S-转移酶π1（GSTP1）/丝裂原活化蛋白激酶（MAPK）轴为潜在靶通路。本研究还通过特异性GSTP1抑制剂开展体外挽救实验，以验证法尼醇是否主要通过调控GSTP1发挥治疗作用。 结果：实验结果证实，法尼醇在体外与体内模型中均可显著缓解肌腱异位骨化。转录组学与网络药理学联合分析显示，法尼醇通过靶向GSTP1/MAPK信号轴发挥作用。特异性GSTP1抑制剂的体外实验进一步验证了上述结论，该抑制剂可逆转法尼醇对成骨过程的抑制作用。 结论：法尼醇通过靶向GSTP1/MAPK通路，抑制创伤后肌腱异位骨化的进展。本研究结果为法尼醇作为异位骨化治疗药物的应用潜力提供了新的理论依据。 整体实验设计：本研究在大鼠肌腱源性干细胞成骨分化第3天，对对照组与法尼醇处理组样本开展RNA测序分析。