中国组织工程研究

中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (5): 1003-1012.doi: 10.12307/2025.282

• 软骨组织构建 cartilage tissue construction • 上一篇    下一篇

二甲双胍抑制PI3K/AKT/mTOR信号通路保护骨关节炎模型大鼠关节软骨

徐田杰1，樊佳欣1，郭小玲1，贾  祥1，赵兴旺2，刘凯楠3，王  茜1   

  1. 1华北理工大学基础医学院，河北省唐山市  063000；2华北理工大学附属医院骨科，河北省唐山市  063000；3邢台医学高等专科学校，河北省邢台市  054000

  • 收稿日期:2023-12-25 接受日期:2024-02-29 出版日期:2025-02-18 发布日期:2024-06-04
  • 通讯作者: 王茜，博士，副教授，硕士生导师，华北理工大学基础医学院，河北省唐山市 063000
  • 作者简介:徐田杰，男，1999年生，陕西省汉中市人，汉族，硕士，主要从事骨组织工程研究。

Metformin exerts a protective effect on articular cartilage in osteoarthritis rats by inhibiting the PI3K/AKT/mTOR pathway 

Xu Tianjie1, Fan Jiaxin1, Guo Xiaoling1, Jia Xiang1, Zhao Xingwang2, Liu kainan3, Wang Qian1   

  1. 1School of Basic Medical Sciences, North China University of Science and Technology, Tangshan 063000, Hebei Province, China; 2Department of Orthopedics, Affiliated Hospital of North China University of Science and Technology, Tangshan 063000, Hebei Province, China; 3College of Basic Medicine, Xingtai Medical College, Xingtai 054000, Hebei Province, China
  • Received:2023-12-25 Accepted:2024-02-29 Online:2025-02-18 Published:2024-06-04
  • Contact: Wang Qian, MD, Associate professor, Master’s supervisor, School of Basic Medical Sciences, North China University of Science and Technology, Tangshan 063000, Hebei Province, China
  • About author:Xu Tianjie, Master, School of Basic Medical Sciences, North China University of Science and Technology, Tangshan 063000, Hebei Province, China

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摘要：



文题释义：

关节软骨：是一层特殊结缔组织，由软骨细胞和细胞外基质组成。关节软骨具有减少摩擦、吸收冲击和分散压力的重要功能，其健康对于关节的正常运动和功能至关重要，然而由于年龄、创伤或疾病等因素的影响，关节软骨可能受到损伤和退化。
PI3K/AKT/mTOR信号通路：是一个重要的细胞信号传导通路，起始于磷脂酰肌醇3-激酶(PI3K)的活化，PI3K会催化磷酸化磷脂酰肌醇二磷酸(PIP2)生成磷脂酰肌醇三磷酸(PIP3)。PIP3进一步激活蛋白激酶B(AKT)，促使AKT被磷酸化，磷酸化的AKT在细胞内激活下游靶蛋白mTOR，从而参与多个细胞生理和病理过程的调控。

背景：研究表明，二甲双胍具有抗炎、抗肿瘤、抗衰老与血管保护作用，可抑制骨关节炎的进展，但其具体的作用机制仍不明确。
目的：探讨二甲双胍对骨关节炎模型大鼠软骨保护的作用机制。
方法：取40只雄性SD大鼠，采用随机数字表法分4组(n=10)：空白组不进行任何手术，假手术组暴露关节腔，模型组、二甲双胍组采用改良Hulth法建立骨关节炎模型；造模后1 d，二甲双胍组大鼠灌胃给予二甲双胍200 mg/(kg•d)，模型组、空白组、假手术组灌胃给予生理盐水，连续给药4周。给药结束后，苏木精-伊红、甲苯胺蓝和番红O-固绿染色观察大鼠膝关节软骨病理形态，免疫组化染色与Western blotting检测大鼠软骨组织中SOX9、Ⅱ型胶原、ADAMTS5、Beclin1、P62、p-PI3K、PI3K、p-AKT、AKT、p-mTOR、mTOR的蛋白表达。 
结果与结论：①苏木精-伊红、甲苯胺蓝和番红O-固绿染色结果显示，空白组、假手术组大鼠膝关节软骨表面光滑，组织形态正常；模型组大鼠膝关节软骨表面不规则，软骨组织出现缺损，软骨细胞数量减少， 软骨基质中蛋白多糖含量减少；相较于模型组，二甲双胍组大鼠膝关节软骨结构损伤有明显改善，软骨表面趋于平整，软骨细胞数量与软骨基质中蛋白多糖含量增加；②免疫组化染色与Western blotting检测结果显示，与空白组、假手术组比较，模型组大鼠软骨组织中SOX9、Ⅱ型胶原、Beclin1蛋白表达降低(P < 0.05)，ADAMTS5、P62及p-PI3K、p-AKT、p-mTOR蛋白表达升高(P < 0.05)；与模型组比较，二甲双胍组大鼠软骨组织中SOX9、Ⅱ型胶原、Beclin1蛋白表达升高(P < 0.05)，ADAMTS5、P62及p-PI3K、p-AKT、p-mTOR蛋白表达降低(P < 0.05)；③结果表明，二甲双胍可通过抑制PI3K/AKT/mTOR信号通路的活化提高骨关节炎模型大鼠软骨细胞自噬活性、减少软骨基质降解，进而发挥关节软骨保护作用。
https://orcid.org/0009-0005-8196-6337(徐田杰)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程


关键词: 骨关节炎, 关节软骨, 自噬, 二甲双胍, PI3K/AKT/mTOR信号通路

Abstract: BACKGROUND: Studies have shown that metformin has anti-inflammatory, anti-tumor, anti-aging and vasoprotective effects, and can inhibit the progression of osteoarthritis, but its specific mechanism of action remains unclear.
OBJECTIVE: To investigate the mechanism of metformin on cartilage protection in a rat model of osteoarthritis.
METHODS: Forty male Sprague-Dawley rats were randomly divided into four groups (n=10 per group): blank, control, sham-operated, and metformin groups. The blank group did not undergo any surgery. In the sham-operated group, the joint cavity was exposed. In the model group and the metformin group, the modified Hulth method was used to establish the osteoarthritis model. At 1 day after modeling, the rats in the metformin group were given 200 mg/kg/d 
metformin by gavage, and the model, blank, and sham-operated groups were given normal saline by gavage. Administration in each group was given for 4 weeks consecutively. Hematoxylin-eosin staining, toluidine blue staining, and safranin O-fast green staining were used to observe the morphological structure of rat knee joints. Immunohistochemical staining and western blot were used to detect the protein expression of SOX9, type II collagen, a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5), Beclin1, P62, phosphatidylinositol 3-kinase (PI3K), p-PI3K, protein kinase B (AKT), p-AKT, mammalian target of rapamycin (mTOR), and p-mTOR in rat cartilage tissue.
RESULTS AND CONCLUSION: The results of hematoxylin-eosin, toluidine blue and safranin O-fast green staining showed smooth cartilage surface of the knee joints and normal histomorphology in the blank group and the sham-operated group, while in the model group, there was irregular cartilage surface of the knee joint and cartilage damage, with a decrease in the number of chondrocytes and the content of proteoglycans in the cartilage matrix. In the metformin group, there was a significant improvement in the damage to the structure of the cartilage in the knee joints of the rats, and the cartilage surface tended to be smooth, with an increase in the number of chondrocytes and the content of proteoglycans in the cartilage matrix. Immunohistochemistry staining and western blot results showed that compared with the control and sham-operated groups, the expression of SOX9, type II collagen, and Beclin1 proteins in the cartilage tissue of rats in the model group was significantly decreased (P < 0.05). Conversely, the expression of ADAMTS5, P62, as well as p-PI3K, p-AKT, and p-mTOR proteins was significantly increased (P < 0.05). Furthermore, compared with the model group, the expression of SOX9, type II collagen, and Beclin1 proteins in the cartilage tissue of rats in the metformin group was significantly increased (P < 0.05), while the expression of ADAMTS5, P62, as well as p-PI3K, p-AKT, and p-mTOR proteins was significantly decreased (P < 0.05). To conclude, Metformin can improve the autophagy activity of chondrocytes and reduce the degradation of cartilage matrix in osteoarthritis rats by inhibiting the activation of PI3K/AKT/mTOR signaling pathway, thus exerting a protective effect on articular cartilage.


Key words: osteoarthritis, articular cartilage, autophagy, metformin, PI3K/AKT/mTOR signaling pathway

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