中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (28): 7251-7259.doi: 10.12307/2026.833

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

少阳生骨方抑制氧化应激延缓膝骨关节炎大鼠软骨衰老

雍  侨1，2，孙  鑫1，2，汪国友1，张  磊1，沈骅睿1，刘  欢1，关钛元1   

  1. 1西南医科大学附属中医医院，四川省泸州市  646000；2西南医科大学中西医结合学院，四川省泸州市  646000
  • 收稿日期:2025-08-04 修回日期:2025-12-31 出版日期:2026-10-08 发布日期:2026-02-09
  • 通讯作者: 关钛元，博士，副教授，西南医科大学附属中医医院，四川省泸州市 646000
  • 作者简介:雍侨，男，2000年生，四川省南充市人，汉族，西南医科大学在读硕士，主要从事骨关节相关研究。
  • 基金资助:

    四川省中医药管理局资助项目(2020JC0148)，项目负责人：关钛元；泸州市科技局项目(2022-SYF-68)，项目负责人：关钛元

Shaoyang Shenggu Fang inhibits oxidative stress and delays cartilage aging in rats with knee osteoarthritis

Yong Qiao1, 2, Sun Xin1, 2, Wang Guoyou1, Zhang Lei1, Shen Huarui1, Liu Huan1, Guan Taiyuan1#br#

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  1. 1Southwest Medical University Affiliated Hospital of Traditional Chinese Medicine, Luzhou 646000, Sichuan Province, China; 2College of Integrated Traditional Chinese and Western Medicine, Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Received:2025-08-04 Revised:2025-12-31 Online:2026-10-08 Published:2026-02-09
  • Contact: Guan Taiyuan, PhD, Associate professor, Southwest Medical University Affiliated Hospital of Traditional Chinese Medicine, Luzhou 646000, Sichuan Province, China
  • About author:Yong Qiao, MS candidate, Southwest Medical University Affiliated Hospital of Traditional Chinese Medicine, Luzhou 646000, Sichuan Province, China; College of Integrated Traditional Chinese and Western Medicine, Southwest Medical University, Luzhou 646000, Sichuan Province, China
  • Supported by:
    a grant from Sichuan Provincial Administration of Traditional Chinese Medicine, No. 2020JC0148 (to GTY); Luzhou Municipal Science and Technology Bureau Project, No. 2022-SYF-68 (to GTY)

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摘要：



文题释义：
少阳生骨方：该方脱胎于经典名方小柴胡汤，并结合《内经》“少阳主骨”理论进行化裁，同时融入了全国名中医孙同郊教授治疗骨病的临床经验。为了增强原方强筋健骨的作用，特加骨碎补与川牛膝两药。最终方剂由柴胡、半夏、党参、黄芩、大枣、甘草、骨碎补、川牛膝8味中药配伍而成，具有和解少阳、通利枢机、补肝益肾、活血止痛的功效，对于治疗膝骨关节炎疗效甚佳。
Wnt/β-catenin通路：在调控膝骨关节炎的发病机制中具有重要作用，可以通过调控抗氧化基因、与氧化应激双向交互，在氧化损伤与细胞死亡中扮演枢纽角色。

背景：前期研究证实，少阳生骨方可以减轻关节软骨退变，促进软骨修复，但其改善膝骨关节炎症状的具体机制尚未明确。Wnt/β-catenin通路与氧化应激在维持关节软骨稳态方面发挥着重要作用。
目的：进一步探讨少阳生骨方调控Wnt/β-catenin通路抑制软骨氧化应激水平延缓大鼠膝骨关节炎软骨衰老的分子作用机制。
方法：32只SD大鼠随机分为空白组、模型组、西药组和中药组4组，除空白组外，其他3组大鼠均采用双膝前交叉韧带离断+内侧半月板前角切除建立膝骨关节炎模型。造模28 d后中药组大鼠予以浓缩少阳生骨方16 g/(kg·d)灌胃给药治疗，西药组大鼠予以盐酸氨基葡萄糖溶液组4 mL/d灌胃，空白组与模型组大鼠给予相同体积生理盐水灌胃。4周后应用苏木精-伊红染色和番红O-固绿染色观察大鼠膝关节软骨损伤退变程度，ELISA法检测血清中相关炎症因子和氧化应激指标水平；采用Western Blot检测膝关节软骨中p21Cip1、p16INK4a蛋白及Wnt信号通路相关蛋白的表达。
结果与结论：①与模型组比较，西药组与中药组大鼠软骨缺损、软骨层变薄、密度下降情况明显改善，Mankin评分显著降低(P < 0.05)；②与模型组相比，西药组和中药组大鼠的血清白细胞介素1β、肿瘤坏死因子α、白细胞介素6水平显著降低(P < 0.05)，超氧化物歧化酶和谷胱甘肽过氧化物酶水平升高，丙二醛浓度降低(均P < 0.05)；中药组p21Cip1、p16INK4a、Wnt5a蛋白表达水平显著下降(P < 0.05与P < 0.01)，β-catenin和C-Myc蛋白表达水平下降(P < 0.05)，糖原合成酶激酶3β蛋白表达水平显著提升(P < 0.05)；③结果提示，少阳生骨方可显著减轻大鼠膝骨关节炎的炎症并缓解软骨衰老，其潜在机制可能是通过少阳生骨方调控Wnt/β-catenin通路抑制软骨氧化应激水平。
https://orcid.org/0009-0000-7038-4095(雍侨)


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: 膝骨关节炎, 少阳生骨方, 氧化应激, 炎症因子, Wnt/β-catenin信号通路, 软骨细胞衰老, 大鼠

Abstract: BACKGROUND: Preliminary studies have demonstrated that Shaoyang Shengguo Fang can alleviate joint cartilage degeneration and promote cartilage repair, but its specific mechanism for alleviating knee osteoarthritis symptoms remains unclear. The Wnt/β-catenin pathway and oxidative stress play crucial roles in maintaining articular cartilage homeostasis.
OBJECTIVE: To investigate the molecular mechanisms by which Shaoyang Shenggu Fang regulates the Wnt/β-catenin pathway to inhibit oxidative stress in cartilage and thereby delay cartilage aging in a rat model of knee osteoarthritis.
METHODS: Thirty-two Sprague-Dawley rats were randomly divided into four groups: a blank control group, a model group, a Western medicine group, and a Chinese medicine group. Animal models of knee osteoarthritis were established in all groups except for the blank control group by transecting the anterior cruciate ligament and resecting the anterior horn of the medial meniscus. At 28 days after modeling, the Chinese medicine group was administered a concentrated Shaoyang Shenggu Fang at 16 g/(kg·d) by gavage, the western medicine group was administered glucosamine hydrochloride solution at 4 mL/d by gavage, and the blank control group and model group were administered the same volume of normal saline by gavage. Four weeks after the start of treatment, hematoxylin-eosin staining and safranin O-fast green staining were used to assess the degree of cartilage injury and degeneration in the rat knee joints. ELISA was used to measure the levels of inflammatory factors and oxidative stress markers in rat serum. Western blot assay was used to determine the expression levels of p21Cip1, p16INK4a proteins, and Wnt signaling pathway-related proteins in rat knee cartilage.
RESULTS AND CONCLUSION: (1) Compared with the model group, the Western medicine group and the Chinese medicine group showed significant improvement in cartilage defects, cartilage layer thinning, and density decrease in rats, with a significant reduction in Mankin scores (P < 0.05). (2) Compared with the model group, the serum levels of interleukin-1β, tumor necrosis factor-α, and interleukin-6 in the Western medicine group and  Chinese medicine group were significantly reduced (P < 0.05), the serum levels of superoxide dismutase and glutathione peroxidase were significantly increased, and malondialdehyde content was significantly decreased (all P < 0.05). In the Chinese medicine group, the expression levels of p21Cip1, p16INK4a, and Wnt5a proteins in the Chinese medicine group were significantly decreased (P < 0.05 and P < 0.01), the expression levels of β-catenin and C-Myc protein expression levels were decreased (P < 0.05), while the expression level of glycogen synthase kinase 3β protein significantly increased (P < 0.05). (3) To conclude, Shaoyang Shenggu Fang can significantly improve the inflammation and cartilage aging in rats with knee osteoarthritis, and its mechanism may be through the Wnt/β-catenin pathway to inhibit oxidative stress levels in cartilage and delay articular cartilage aging.

Key words: knee osteoarthritis, Shaoyang Shenggu Fang, oxidative stress, inflammatory factors, Wnt/β-catenin signaling pathway, chondrocyte aging, rats

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