中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (19): 4825-4835.doi: 10.12307/2026.245

• 骨髓干细胞 bone marrow stem cells •    下一篇

五加皮外泌体样纳米囊泡促进人骨髓间充质干细胞成骨分化

钟焯岚1，彭芝娜1，田晓红2，韩翠菲3，张紫菡3，楚佳奇4   

  1. 1广东医科大学第一临床医学院，广东省湛江市   524001；2河北中石油中心医院，河北省廊坊市   065000；3广东医科大学，广东省湛江市   524003；4广东医科大学附属医院，广东省湛江市   524001
  • 收稿日期:2025-08-08 接受日期:2025-12-10 出版日期:2026-07-08 发布日期:2026-02-13
  • 通讯作者: 楚佳奇，副研究员，广东医科大学附属医院，广东省湛江市 524001
  • 作者简介:钟焯岚，男，1997年生，广东省江门市人，汉族，广东医科大学在读硕士，主要从事植物源性纳米囊泡改善骨质疏松方面的研究。
  • 基金资助:
    广东医科大学附属医院“临床医学”科技共建平台项目(CLP2021A001)，项目负责人：楚佳奇

Acanthopanax exosome-like nanovesicles promote osteogenic differentiation of human bone marrow mesenchymal stem cells

Zhong Zhuolan1, Peng Zhina1, Tian Xiaohong2, Han Cuifei3, Zhang Zihan3, Chu Jiaqi4   

  1. 1First Clinical Medical College of Guangdong Medical University, Zhanjiang 524001, Guangdong Province, China; 2Hebei PetroChina Central Hospital, Langfang 065000, Hebei Province, China; 3Guangdong Medical University, Zhanjiang 524003, Guangdong Province, China; 4Affiliated Hospital of Guangdong Medical University, Zhanjiang 524001, Guangdong Province, China
  • Received:2025-08-08 Accepted:2025-12-10 Online:2026-07-08 Published:2026-02-13
  • Contact: Chu Jiaqi, Associate researcher, Affiliated Hospital of Guangdong Medical University, Zhanjiang 524001, Guangdong Province, China
  • About author:Zhong Zhuolan, MS candidate, First Clinical Medical College of Guangdong Medical University, Zhanjiang 524001, Guangdong Province, China
  • Supported by:
    Guangdong Medical University Affiliated Hospital "Clinical Medicine" Science and Technology Co-construction Platform Project, No. CLP2021A001 (to CJQ) 

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摘要：

文题释义：

五加皮来源外泌体样纳米囊泡：采用差速离心结合蔗糖梯度离心技术，从中药五加皮中分离得到富含蛋白质、microRNA、mRNA及代谢物等生物活性成分的纳米级囊泡。此类囊泡粒径集中在100-200 nm之间，具有天然双层膜结构，能够被靶细胞摄取，进而调控细胞增殖、分化及功能状态。
转化生长因子β1/Smad2/3信号通路：此通路在骨代谢调控中发挥关键的信号传导作用。转化生长因子β1配体通过结合Ⅱ型受体并激活Ⅰ型受体，促进下游Smad2/3蛋白磷酸化，并形成转录复合物进入细胞核，调控成骨相关基因(如Runt相关转录因子2、骨钙素)的表达。在成骨过程中，此通路在不同阶段表现出动态调控特性，既可促进骨祖细胞增殖与早期分化，又可在后期阶段调控矿化及骨重塑。

摘要
背景：五加皮及其提取物具有促成骨作用，但五加皮来源外泌体样纳米囊泡的成骨效应及机制尚未阐明。
目的：探讨五加皮来源外泌体样纳米囊泡促进人骨髓间充质干细胞成骨分化的分子机制及对骨质疏松症的预防作用。
方法：①采用梯度密度离心法提取人骨髓间充质干细胞；采用差速离心、蔗糖梯度密度离心法分离五加皮来源外泌体样纳米囊泡；②以0，2.5，5 μg/mL五加皮来源外泌体样纳米囊泡干预人骨髓间充质干细胞，采用碱性磷酸酶染色、茜素红染色、qRT-PCR及Western blot检测成骨分化水平；③转录组测序筛选关键通路并用转化生长因子β1受体抑制剂验证；④构建卵巢切除骨质疏松大鼠模型，腹腔注射五加皮来源外泌体样纳米囊泡干预12周，Micro-CT分析骨微结构，组织学染色检测成骨蛋白表达。

结果与结论：①五加皮来源外泌体样纳米囊泡呈典型的杯托状或圆盘形；②体外实验证实，五加皮来源外泌体样纳米囊泡剂量依赖性地促进骨髓间充质干细胞成骨分化，表现为碱性磷酸酶活性升高、矿化结节增多及成骨基因表达上调；③转录组分析显示，五加皮来源外泌体样纳米囊泡通过激活转化生长因子β1/Smad2/3信号通路，上调转化生长因子β1及磷酸化Smad2/3蛋白表达，且转化生长因子β1受体抑制剂可部分抑制它的促成骨效应；④动物实验结果显示，5 mg/kg五加皮来源外泌体样纳米囊泡干预后，卵巢切除大鼠骨密度、骨体积分数及骨小梁厚度均显著增加(P < 0.05)，胶原纤维生成明显，且骨组织中Runt相关转录因子2、骨钙素及转化生长因子β1蛋白表达上调；主要脏器组织学观察未见明显毒性变化。结果表明，五加皮来源外泌体样纳米囊泡通过激活转化生长因子β1/Smad2/3通路促进人骨髓间充质干细胞成骨分化，并有效改善骨质疏松。

https://orcid.org/0009-0006-6995-9828 (钟焯岚) 


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: 五加皮, 外泌体样纳米囊泡, 骨髓间充质干细胞, 转化生长因子β1信号通路, Smad2/3, 成骨分化, 骨质疏松, 转录组测序, 工程化干细胞

Abstract: BACKGROUND:  Acanthopanax and its extracts exhibit osteogenic effects, but the osteogenic potential and mechanisms of acanthopanax exosome-like nanovesicles remain unclear. 
OBJECTIVE: To investigate the molecular mechanism by which acanthopanax exosome-like nanovesicles promote osteogenic differentiation of human bone marrow mesenchymal stem cells and their preventive role in osteoporosis.
METHODS: (1) Human bone marrow mesenchymal stem cells were extracted by gradient density centrifugation. Exosome-like nanovesicles derived from acanthopanax were isolated by differential centrifugation and sucrose gradient density centrifugation. (2) Human bone marrow mesenchymal stem cells were treated with 0, 2.5, and 5 μg/mL acanthopanax exosome-like nanovesicles. Osteogenic differentiation was assessed by alkaline phosphatase staining, Alizarin red staining, qRT-PCR, and western blot assay. (3) Key pathways were identified by transcriptome sequencing and validated with a transforming growth factor β1 receptor inhibitor. (4) An ovariectomized rat model of osteoporosis was established. After 12 weeks of intraperitoneal injection of exosome-like nanovesicles derived from acanthopanax, bone microstructure was analyzed by micro-CT and osteogenic protein expression was assessed by histological staining. 
RESULTS AND CONCLUSION: (1) Acanthopanax exosome-like nanovesicles exhibited a cup-shaped or discoid morphology. (2) In vitro, acanthopanax exosome-like nanovesicles dose-dependently promoted osteogenic differentiation of bone marrow mesenchymal stem cells, as evidenced by increased alkaline phosphatase activity, enhanced mineralization nodule formation, and upregulated osteogenic gene expression. (3) Transcriptome analysis revealed that exosome-like nanovesicles activated the transforming growth factor-β1/Smad2/3 pathway, upregulating transforming growth factor-β1 and phosphorylated Smad2/3 protein expression. Moreover, the transforming growth factor-β1 receptor inhibitor partially inhibited the osteogenic effects. (4) Animal experimental results demonstrated that treatment with 5 mg/kg exosome-like nanovesicles from acanthopanax significantly increased bone mineral density, bone volume fraction, and trabecular thickness in ovariectomized rats (P < 0.05), significantly enhanced collagen fibrillogenesis, and upregulated the expression of Runt-related transcription factor 2, osteocalcin, and transforming growth factor-β1 proteins in bone tissue. No significant toxicity was observed in major organ histological observations. Results showed that exosome-like nanovesicles derived from acanthopanax promote osteogenic differentiation of human bone marrow mesenchymal stem cells by activating the transforming growth factor β1/Smad2/3 pathway and effectively improve osteoporosis.

Key words: acanthopanax, exosome-like nanovesicles, bone marrow mesenchymal stem cells, transforming growth factor-β1 signaling pathway, Smad2/3, osteogenic differentiation, osteoporosis, transcriptome sequencing, engineered stem cells

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