中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (25): 6433-6445.doi: 10.12307/2026.455

• 骨组织构建 bone tissue construction •    下一篇

10-羟基-2-癸烯酸介导自噬与抗氧化能力增强促成骨分化

胡  杰1，2，何  辉1，2，麻凤玉1，2，沈孝天1，2，袁章琴1，2，梁  婷1，2，韩凤选1，2   

  1. 1苏州大学附属第一医院骨科，江苏省苏州市   215000；2苏州大学苏州医学院骨科研究所，江苏省苏州市   215000
  • 收稿日期:2025-09-21 修回日期:2026-02-05 出版日期:2026-09-08 发布日期:2026-04-17
  • 通讯作者: 韩凤选，博士，教授，苏州大学附属第一医院骨科，江苏省苏州市 215000；苏州大学苏州医学院骨科研究所，江苏省苏州市 215000
  • 作者简介:第一作者：胡杰，男，1999年生，江西省吉安市人，汉族，硕士，主要从事干细胞和组织再生相关研究。 共同第一作者：何辉，女，2002年生，山东省菏泽市人，汉族，硕士，主要从事干细胞和组织再生相关研究。
  • 基金资助:
    江苏省基础研究计划自然科学基金-杰出青年基金项目(BK20240020)，项目负责人：韩凤选 

10-Hydroxy-2-decenoic acid facilitates osteogenic differentiation via the enhancement of autophagy and antioxidant capacity

Hu Jie1, 2, He Hui1, 2, Ma Fengyu1, 2, Shen Xiaotian1, 2, Yuan Zhangqin1, 2, Liang Ting1, 2, Han Fengxuan1, 2   

  1. 1Department of Orthopedic Surgery, First Affiliated Hospital, Soochow University, Suzhou 215000, Jiangsu Province, China; 2Orthopedic Institute, Suzhou Medical College, Soochow University, Suzhou 215000, Jiangsu Province, China
  • Received:2025-09-21 Revised:2026-02-05 Online:2026-09-08 Published:2026-04-17
  • Contact: Han Fengxuan, PhD, Professor, Department of Orthopedic Surgery, First Affiliated Hospital, Soochow University, Suzhou 215000, Jiangsu Province, China; Orthopedic Institute, Suzhou Medical College, Soochow University, Suzhou 215000, Jiangsu Province, China
  • About author:1Department of Orthopedic Surgery, First Affiliated Hospital, Soochow University, Suzhou 215000, Jiangsu Province, China; 2Orthopedic Institute, Suzhou Medical College, Soochow University, Suzhou 215000, Jiangsu Province, China Hu Jie, MS, Department of Orthopedic Surgery, First Affiliated Hospital, Soochow University, Suzhou 215000, Jiangsu Province, China; Orthopedic Institute, Suzhou Medical College, Soochow University, Suzhou 215000, Jiangsu Province, China He Hui, MS, Department of Orthopedic Surgery, First Affiliated Hospital, Soochow University, Suzhou 215000, Jiangsu Province, China; Orthopedic Institute, Suzhou Medical College, Soochow University, Suzhou 215000, Jiangsu Province, China Hu Jie and He Hui contributed equally to this work.
  • Supported by:

    Jiangsu Provincial Natural Science Foundation for Basic Research - Outstanding Young Scientist Fund Project, No. BK20240020 (to HFX) 


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摘要：



文题释义：
10-羟基-2-癸烯酸：是一种来源于蜂王浆的不饱和脂肪酸，具有较强的抗炎、抗氧化和免疫调节作用，对类风湿关节炎、神经炎症、肿瘤、肌肉萎缩、细菌感染等疾病具有潜在治疗价值。
自噬：是真核细胞在自噬相关基因调控下利用溶酶体降解自身细胞质蛋白和受损细胞器的过程，是维持干细胞稳态和功能的重要机制。自噬包括生理条件下的基础型自噬和应激条件下的诱导型自噬。

背景：10-羟基-2-癸烯酸具有较强的抗炎、抗氧化和免疫调节作用，但它在骨代谢调控中的作用尚不明确。
目的：探讨10-羟基-2-癸烯酸在骨重建过程中的调节作用及潜在机制。
方法：①采用不同浓度(0，0.5，1，2，4 mmol/L)的10-羟基-2-癸烯酸培养大鼠骨髓间充质干细胞，细胞骨架染色评估细胞形态，活死染色和CCK-8法检测细胞活性、增殖情况；②采用不同浓度(0，0.5，1，2 mmol/L)的10-羟基-2-癸烯酸培养大鼠骨髓间充质干细胞，成骨诱导分化后，碱性磷酸酶染色和茜素红染色评估成骨分化情况，Western blot和免疫荧光染色分析成骨相关蛋白表达；③诱导小鼠骨髓单个核细胞分化为巨噬细胞，分别加入含不同浓度(0，0.5，1，2 mmol/L)10-羟基-2-癸烯酸的破骨诱导分化培养基，抗酒石酸酸性磷酸酶和F-actin染色检测破骨细胞生成情况；④将大鼠骨髓间充质干细胞饥饿培养6 h后常规培养，并分组：对照组(不进行任何干预)、10-羟基-2-癸烯酸组、10-羟基-2-癸烯酸+ AS1842856组、10-羟基-2-癸烯酸 +EX-527组，10-羟基-2-癸烯酸干预浓度为0.5 mmol/L，AS1842856为叉头框转录因子O1抑制剂，EX-527为沉默调节蛋白1抑制剂，Western blot和免疫荧光染色分析沉默调节蛋白1/叉头框转录因子O1通路激活情况以及自噬、成骨相关蛋白表达；⑤将大鼠骨髓间充质干细胞分组培养：对照组、H2O2组、H2O2+10-羟基-2-癸烯酸组，10-羟基-2-癸烯酸干预浓度为0.5 mmol/L，H2O2干预24 h后进行对应的药物干预，成骨诱导后分别进行碱性磷酸酶与茜素红染色；⑥将大鼠骨髓间充质干细胞分5组培养：对照组、H2O2组、H2O2+10-羟基-2-癸烯酸组、H2O2+10-羟基-2-癸烯酸+AS1842856组、H2O2+10-羟基-2-癸烯酸+EX-527组，10-羟基-2-癸烯酸干预浓度为0.5 mmol/L，H2O2干预24 h后进行对应的药物干预，Western blot检测沉默调节蛋白1/叉头框转录因子O1信号通路与抗氧化相关蛋白表达，TUNEL染色和β-半乳糖苷酶染色评估细胞凋亡和衰老情况。
结果与结论： ①细胞骨架染色、活死染色和CCK-8检测显示，0.5，1，2 mmol/L 10-羟基-2-癸烯酸可促进大鼠骨髓间充质干细胞的增殖，后续实验选择该3种干预浓度；②碱性磷酸酶、茜素红染色与Western blot和免疫荧光染色分析显示，0.5 mmol/L 10-羟基-2-癸烯酸可促进大鼠骨髓间充质干细胞的成骨分化与矿化，提升成骨相关蛋白表达；③抗酒石酸酸性磷酸酶和F-actin染色显示，0.5 mmol/L 10-羟基-2-癸烯酸可显着抑制破骨细胞的生成；④Western blot和免疫荧光染色显示，10-羟基-2-癸烯酸可激活沉默调节蛋白1/叉头框转录因子O1信号通路促进叉头框转录因子O1去乙酰化及其核转位，上调自噬相关蛋白和抗氧化酶的表达；⑤碱性磷酸酶与茜素红染色显示，在氧化应激状态下，10-羟基-2-癸烯酸可促进大鼠骨髓间充质干细胞的成骨分化与矿化；⑥Western blot检测显示，在氧化应激状态下，10-羟基-2-癸烯酸可通过激活沉默调节蛋白1/叉头框转录因子O1信号通路增强大鼠骨髓间充质干细胞的抗氧化能力。 TUNEL染色和β-半乳糖苷酶染色显示，在氧化应激状态下，10-羟基-2-癸烯酸可通过激活沉默调节蛋白1/叉头框转录因子O1信号通路减少大鼠骨髓间充质干细胞的凋亡与衰老。结果表明，10-羟基-2-癸烯酸通过调控沉默调节蛋白1/叉头框转录因子O1信号通路增强自噬与抗氧化能力，进而促进成骨分化。
https://orcid.org/0009-0007-7881-2200 (胡杰)  


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 骨质疏松, 骨髓间充质干细胞, 10-羟基-2-癸烯酸, 成骨分化, 破骨细胞, 沉默调节蛋白1, 自噬, 抗氧化

Abstract:

BACKGROUND: 10-Hydroxy-2-decenoic acid (10-HDA) exhibits potent anti-inflammatory, antioxidant, and immunomodulatory effects, but its role in regulating bone metabolism remains unclear.
OBJECTIVE: To investigate the regulatory effects and underlying mechanisms of 10-HDA on bone remodeling.
METHODS: (1) Bone marrow-derived mesenchymal stem cells were isolated from rats and treated with different concentrations of 10-HDA (0, 0.5, 1, 2, and 4 mmol/L). Cytoskeletal staining was used to assess cell morphology, while live/dead staining and the cell counting kit-8 assay were employed to evaluate cell viability and proliferation. (2) Bone marrow-derived mesenchymal stem cells were isolated from rats and treated with different concentrations of 10-HDA (0, 0.5, 1, and 2 mmol/L). After osteogenic induction, alkaline phosphatase staining and alizarin red staining were conducted to assess osteogenic differentiation. The expression of osteogenesis-related proteins was analyzed by western blot and immunofluorescence staining. (3) Mouse bone marrow mononuclear cells were induced to differentiate into macrophages by adding osteoclast-inducing differentiation medium containing different concentrations of 10-HDA (0, 0.5, 1, and 
2 mmol/L). Tartrate-resistant acid phosphatase staining and F-actin staining were used to evaluate osteoclast formation. (4) Rat bone marrow mesenchymal stem cells were starved for 6 hours before conventional culture and grouped as follows: control group (no intervention), 10-HDA group, 10-HDA + AS1842856 group, 10-HDA + EX-527 group. 10-Hydroxy-2-decenoic acid was administered at a concentration of 0.5 mmol/L. AS1842856 was a forkhead box transcription factor O1 inhibitor, and EX-527 was a silent information regulator 1 inhibitor. Furthermore, activation of the silent information regulator 1/forkhead box transcription factor O1 signaling pathway and expression of autophagy- and antioxidant-related proteins were assessed by western blot and immunofluorescence. (5) Rat bone marrow mesenchymal stem cells were cultured in five groups: control group, H₂O₂ group, and H₂O₂ + 10-HDA group. The intervention concentration of 10-HDA was 0.5 mmol/L. Corresponding drug interventions were administered 24 hours after H₂O₂ exposure. After osteogenic induction, alkaline phosphatase staining and alizarin red staining were conducted. (6) Rat bone marrow mesenchymal stem cells were cultured in five groups: control group, H₂O₂ group, H₂O₂+10-HDA group, H₂O₂+10-HDA+AS1842856 group, and H₂O₂+10-HDA+EX-527 group. The intervention concentration of 10-HDA was 0.5 mmol/L. Corresponding drug interventions were administered 24 hours after H₂O₂ exposure. Western blot analysis was performed to detect the expression of silent information regulator 1/forkhead box transcription factor O1 signaling pathway and antioxidant-related proteins. TUNEL staining and β-galactosidase staining were performed to evaluate apoptosis and senescence. 
RESULTS AND CONCLUSION: (1) At the concentration of 0.5, 1, and 2 mmol/L, 10-HDA promoted the proliferation of bone marrow-derived mesenchymal stem cells. Therefore, these three intervention concentrations were selected for subsequent experiments. (2) 10-HDA (0.5 mmol/L) significantly enhanced the osteogenic differentiation and mineralization of bone marrow-derived mesenchymal stem cells and increased the expression of osteogenesis-related proteins, as evidenced by alkaline phosphatase staining, alizarin red staining, western blot and immunofluorescence staining. (3) Tartrate-resistant acid phosphatase staining and F-actin staining revealed that 0.5 mmol/L 10-HDA significantly inhibited osteoclast formation. (4) Western blot and immunofluorescence staining revealed that 10-HDA activated the silent information regulator 1/forkhead box transcription factor O1 signaling pathway to promote deacetylation and nuclear translocation of forkhead box transcription factor O1, thereby upregulating autophagy-related proteins and antioxidant enzymes. (5) Alkaline phosphatase and alizarin red staining revealed that 10-HDA promotes osteogenic differentiation and mineralization in rat bone marrow-derived mesenchymal stem cells under oxidative stress conditions. (6) Western blot analysis revealed that under oxidative stress conditions, 10-HDA enhanced the antioxidant capacity of rat bone marrow mesenchymal stem cells by activating the silent information regulator 1/forkhead box transcription factor O1 signaling pathway. TUNEL and β-galactosidase staining revealed that under oxidative stress conditions, 10-HDA reduced apoptosis and senescence in rat bone marrow mesenchymal stem cells by activating the silent information regulator 1/forkhead box transcription factor O1 signaling pathway. To conclude, 10-HDA enhances autophagy and antioxidant capacity by regulating the silent information regulator 1/forkhead box transcription factor O1 signaling pathway, thereby promoting osteogenic differentiation.
Key words: osteoporosis; bone marrow-derived mesenchymal stem cells; 10-hydroxy-2-decenoic acid; osteogenic differentiation; osteoclasts; silent information regulator 1; autophagy; antioxidant


Key words: osteoporosis, bone marrow-derived mesenchymal stem cells, 10-hydroxy-2-decenoic acid, osteogenic differentiation, osteoclasts, silent information regulator 1, autophagy, antioxidant

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