中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (35): 9151-9158.doi: 10.12307/2026.432

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

考比替尼与达沙替尼联合槲皮素缓解炎症性软骨细胞衰老的比较

赵敏君1，2，吴绪波1，3，尹枧郦1，2，葛杨硕1，2，丁佳滢1，2，黄春萌1，2，4，孟婷婷1，2，4，王学宗5，刘振峰6，丁道芳1，2   

  1. 上海中医药大学，1康复医学院，2康复医学研究所，上海市   201203；3上海市浦东新区人民医院，上海市   201200；4上海市第二康复医院，上海市   200441；5上海中医药大学附属曙光医院石氏伤科医学中心，上海市   200135；6江苏省无锡市第九人民医院，江苏省无锡市   214062
  • 收稿日期:2025-09-11 修回日期:2026-01-19 出版日期:2026-12-18 发布日期:2026-04-25
  • 通讯作者: 丁道芳，博士，研究员，上海中医药大学，康复医学院，康复医学研究所，上海市 201203
  • 作者简介:赵敏君，女，1999年生，云南省红河州人，蒙古族，上海中医药大学康复医学院在读硕士，主要从事骨伤康复的研究。
  • 基金资助:
    国家自然科学基金项目(82174406)，项目负责人：王学宗；国家自然科学基金项目(81902306)，项目负责人：丁道芳；上海市卫生健康委员会传统医学研究项目(2024QN012)，项目负责人：黄春萌；上海中医药大学科学技术发展项目(23KFL023)，项目负责人： 丁道芳；江苏省无锡市科技项目(ZYYB06)，项目负责人：刘振峰

Effects of cobimetinib versus combination of dasatinib and quercetin on stress-induced chondrocyte senescence

Zhao Minjun1, 2, Wu Xubo1, 3, Yin Jianli1, 2, Ge Yangshuo1, 2, Ding Jiaying1, 2, Huang Chunmeng1, 2, 4, Meng Tingting1, 2, 4, Wang Xuezong5, #br# Liu Zhenfeng6, Ding Daofang1, 2#br#   

  1. 1School of Rehabilitation Medicine, 2Institute of Rehabilitation Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; 3Pudong New Area People’s Hospital, Shanghai 201200, China; 4Shanghai Second Rehabilitation Hospital, Shanghai 200441, China; 5Shishi Orthopedic Medical Center, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200135, China; 6The Ninth People’s Hospital of Wuxi City, Wuxi 214062, Jiangsu Province, China
  • Received:2025-09-11 Revised:2026-01-19 Online:2026-12-18 Published:2026-04-25
  • Contact: Ding Daofang, PhD, Researcher, School of Rehabilitation Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Institute of Rehabilitation Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
  • About author:Zhao Minjun, MS candidate, School of Rehabilitation Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Institute of Rehabilitation Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
  • Supported by:
    National Natural Science Foundation of China, Nos. 82174406 (to WXZ) and 81902306 (to DDF); the Traditional Chinese Medicine Research Project of Shanghai Municipal Health Commission, No. 2024QN012 (to HCM); the Scientific and Technological Development Project of Shanghai University of Traditional Chinese Medicine, No. 23KFL023 (to DDF); Wuxi Science and Technology Project, No. ZYYB06 (to LZF)

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摘要：



文题释义：
衰老相关分泌表型(Senescence-associated secretory phenotype，SASP)：由衰老细胞分泌多种生物因子(如细胞因子、趋化因子、生长因子、蛋白酶等)，改变周围微环境进而对邻近细胞及组织产生影响的表型。
细胞衰老：细胞在执行生命活动过程中，随着时间的推移，细胞增殖与分化能力和生理功能逐渐发生衰退的变化过程。

背景：骨关节炎与衰老密切相关，其特征为关节软骨退化、软骨下骨硬化及低度炎症反应。衰老和损伤是促炎因子介导关节病理变化的重要触发因素。考比替尼(Cobimetinib)为MEK1抑制剂，对软骨细胞炎症性衰老的影响尚不清楚。
目的：旨在比较考比替尼与经典抗衰老药物组合即达沙替尼(Dasatinib)和槲皮素(Quercetin)在抑制软骨细胞炎症性衰老方面的作用差异，探索其在骨关节炎治疗中的潜在应用价值。
方法：分离和培养原代软骨细胞，采用CCK-8法检测细胞活力。将细胞分为对照组、模型组、D+Q组、考比替尼组，除对照组细胞外，其余3组细胞使用白细胞介素1β诱导建立软骨细胞炎症性衰老模型，再分别给予考比替尼或达沙替尼联合槲皮素(D+Q组合)干预，使用β-半乳糖苷酶染色观察细胞衰老表型，实时荧光定量PCR、Western Blot和免疫荧光检测MEK-ERK1/2通路的激活情况，P16、P21、P53等衰老标志物及衰老相关分泌表型因子(如诱导型一氧化氮合酶、环氧化酶2、趋化因子3、白细胞介素6)的表达水平，并评估细胞外基质合成与降解相关分子(COL2A1、基质金属蛋白酶13、基质金属蛋白酶3)的变化。
结果与结论：考比替尼和达沙替尼联合槲皮素均能有效抑制MEK-ERK1/2通路的激活，缓解软骨细胞的衰老表型，显著下调P16、P21、P53表达，减少衰老相关分泌表型因子水平，恢复COL2A1表达并抑制基质金属蛋白酶13、基质金属蛋白酶3表达。结果说明，考比替尼通过抑制MEK-ERK1/2通路，有效缓解白细胞介素1β诱导的软骨细胞炎症性衰老，与达沙替尼联合槲皮素组合具有相似的抗衰老作用，同时在促进基质合成、抑制降解方面更具有潜力。

https://orcid.org/0009-0008-3808-6205 (赵敏君)；https://orcid.org/0000-0003-4056-142X (丁道芳)


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: 骨关节炎, 细胞衰老, 软骨细胞, 考比替尼, 达沙替尼, 槲皮素, 衰老相关分泌表型, MEK-ERK1/2信号通路

Abstract: BACKGROUND:  Osteoarthritis is closely related to aging and characterized by degeneration of articular cartilage, subchondral bone sclerosis, and low-grade inflammatory responses. Aging and injury are significant triggers for inflammatory factors that mediate joint pathological changes. Cobimetinib, a MEK1 inhibitor, has an unclear effect on the inflammatory senescence of chondrocytes.
OBJECTIVE: To compare the effects of cobimetinib and the classical senolytic drug combination of dasatinib and quercetin (D+Q) in suppressing inflammation-induced chondrocyte senescence, and to explore their potential therapeutic value in osteoarthritis.
METHODS: Primary chondrocytes were isolated and cultured. Cell viability was assessed by cell counting kit-8 assay. Cells were divided into control, model, D+Q, and cobimetinib groups. Except for the control group, inflammatory senescence chondrocyte models were induced by interleukin-1β in the other three groups. The cobimetinib and D+Q groups were treated with cobimetinib and D+Q, respectively. Senescent phenotypes were evaluated using SA-β-Gal staining. The activation status of the MEK-ERK1/2 pathway, expression levels of senescence markers (P16, P21, P53) and senescence-associated secretory phenotype factors (such as inducible nitric oxide synthase, cyclooxygenase-2, chemokine 3, interleukin-6) were analyzed by real-time quantitative PCR, western blot, and immunofluorescence. Additionally, changes in molecules related to extracellular matrix synthesis and degradation (COL2A1, matrix metalloproteinase-13, and matrix metalloproteinase-3) were assessed.
RESULTS AND CONCLUSION: Both cobimetinib and D+Q could effectively inhibit the activation of the MEK-ERK1/2 pathway, ameliorate the senescent phenotypes of chondrocytes, as evidenced by downregulation of P16, P21, and P53, attenuation of senescence-associated secretory phenotype factor levels, restoration of COL2A1 expression, and suppression of matrix metalloproteinase-13 and matrix metalloproteinase-3. Overall, cobimetinib effectively alleviates interleukin-1β-induced inflammatory senescence in chondrocytes by inhibiting the MEK-ERK1/2 pathway. Compared with D+Q, it exhibits comparable anti-senescent effects while demonstrating greater potential in promoting matrix synthesis and inhibiting degradation.


Key words: osteoarthritis, cellular senescence, chondrocytes, cobimetinib, dasatinib, quercetin, senescence-associated secretory phenotype, MEK-ERK1/2 signaling pathway

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