中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (24): 6214-6219.doi: 10.12307/2026.176

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

右归丸减缓膝骨关节炎模型大鼠软骨细胞焦亡的作用机制

范元赫1，杨永菊2，何璠玙1，乔  隆2，张  宇1，张  帅3，李志文3，关雪峰4   

  1. 1辽宁中医药大学，辽宁省沈阳市   110847；2辽宁中医药大学附属医院，辽宁省沈阳市   110032；3辽宁中医药大学附属第二医院，辽宁省沈阳市   110034；4沈阳药科大学，辽宁省沈阳市   110016
  • 收稿日期:2025-04-16 修回日期:2025-10-08 出版日期:2026-08-28 发布日期:2026-01-29
  • 通讯作者: 关雪峰，博士，教授，主任医师，博士生导师，沈阳药科大学，辽宁省沈阳市 110016
  • 作者简介:范元赫，男，1994年生，辽宁省丹东市人，汉族，2025年辽宁中医药大学毕业，博士，主要从事中医药防治骨关节病研究。
  • 基金资助:
    辽宁省创新能力提升联合基金(2022NLTS-13-02)，项目负责人：张宇；2024年联合基金项目博士科研启动项目(2023BSBA-217)，项目负责人：何璠玙

Mechanism by which Yougui Pill inhibits pyroptosis of chondrocytes in rats with knee osteoarthritis

Fan Yuanhe1, Yang Yongju2, He Fanyu1, Qiao Long2, Zhang Yu1, Zhang Shuai3, Li Zhiwen3, Guan Xuefeng4   

  1. 1Liaoning University of Traditional Chinese Medicine, Shenyang 110847, Liaoning Province, China; 2Liaoning University of Traditional Chinese Medicine Affiliated Hospital, Shenyang 110032, Liaoning Province, China; 3Second Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang 110034, Liaoning Province, China; 4Shenyang Pharmaceutical University, Shenyang 110006, Liaoning Province, China
  • Received:2025-04-16 Revised:2025-10-08 Online:2026-08-28 Published:2026-01-29
  • Contact: Guan Xuefeng, PhD, Professor, Chief physician, Shenyang Pharmaceutical University, Shenyang 110006, Liaoning Province, China
  • About author:Fan Yuanhe, PhD, Liaoning University of Traditional Chinese Medicine, Shenyang 110847, Liaoning Province, China
  • Supported by:
    Liaoning Province Joint Fund for Enhancing Innovation Capacity, No. 2022NLTS-13-02 (to ZY); 2024 Joint Fund Project for Doctoral Research Startup, No. 2023BSBA-217 (to HFY) 

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摘要：



文题释义：
膝骨关节炎：是一种以关节软骨退行性变、骨质增生及滑膜炎症为特征的慢性退行性关节疾病，主要病理表现为软骨厚度减少(正常2-
4 mm，晚期可降至1 mm以下)、关节间隙狭窄(内侧间隙常小于3 mm)、骨赘形成(骨赘大于2 mm)。影像学Kellgren-Lawrence分级是常用诊断标准，其中2级以上(关节间隙明确狭窄伴骨赘形成)具有临床诊断价值。疼痛目测类比评分≥3分、WOMAC指数> 39分提示症状显著，治疗方案需根据K-L分级、疼痛程度及功能受限情况个体化制定。
细胞焦亡：是一种程序性细胞死亡方式，特征为细胞膜穿孔、细胞肿胀并最终裂解，释放大量炎症因子。该过程由gasdermin蛋白介导，其N端结构域在激活后形成直径10-16 nm的膜孔，导致细胞渗透压失衡。与凋亡不同，焦亡伴随白细胞介素1β和白细胞介素18等前炎症因子释放。在骨科领域，焦亡与骨关节炎密切相关。研究显示，骨关节炎患者滑膜中gasdermin D表达量较正常组织升高两三倍，且白细胞介素1β水平可达100-200 pg/mL，显著高于健康对照组(通常< 10 pg/mL)，抑制焦亡通路可能成为治疗骨关节疾病的新靶点。

背景：右归丸出自《景岳全书》，有研究证实右归丸对膝骨关节炎患者疗效显著，但作用机制尚不明确。
目的：探讨右归丸改善大鼠膝骨关节炎的潜在分子作用机制。
方法：48只SPF级SD大鼠采用随机数字表法分为4组，分别为空白组、模型组、右归丸组、塞来昔布组，后3组采用改良Hulth法对大鼠膝关节进行手术造模，伤口愈合后驱赶8周。造模结束后，塞来昔布组大鼠灌胃塞来昔布混悬液，右归丸组大鼠灌胃右归丸水煎液，假手术组与模型组灌胃等体积生理盐水，1次/d，连续灌胃4周。采用苏木精-伊红染色、甲苯胺蓝染色、番红-固绿染色观察大鼠软骨组织病理变化，透射电镜观察大鼠软骨细胞超微结构，ELISA法检测大鼠血清中白细胞介素18、白细胞介素1β、肿瘤坏死因子α水平，Western blot法检测大鼠软骨组织PI3K、AKT、NF-κB、p-PI3K、p-AKT、p-P65、NLRP3、GSDMD、GSDMD-N、Caspase1、Cleaved-Caspase1、白细胞介素18、白细胞介素1β蛋白表达。
结果与结论：①与空白组比较，模型组大鼠软骨边缘严重破坏，软骨组织缺损，软骨层细胞变薄、排列紊乱，软骨下骨增生，潮线紊乱，软骨细胞结构损伤严重、焦亡小体形成，血清肿瘤坏死因子α、白细胞介素18、白细胞介素1β水平显著升高(P < 0.05)，软骨组织p-PI3K、p-AKT、p-P65、NLRP3、GSDMD-N、Cleaved-Caspase1、白细胞介素18、白细胞介素1β蛋白表达均显著升高(P < 0.01)；②与模型组比较，右归丸组和塞来昔布组大鼠软骨结构趋于正常，软骨着色较深，软骨增厚，软骨细胞胞膜较完整，血清中肿瘤坏死因子α、白细胞介素18、白细胞介素1β水平显著减低(P < 0.05)，软骨组织p-PI3K、p-AKT、p-P65、NLRP3、GSDMD-N、Cleaved-Caspase1、白细胞介素18、白细胞介素1β蛋白表达显著降低(P < 0.01)；③与塞来昔布组比较，右归丸组软骨细胞排列较规整，关节软骨表面较光滑，软骨层增厚明显，潮线相对完整，软骨细胞胞膜相对完整，血清中肿瘤坏死因子α、白细胞介素18、白细胞介素1β水平显著减低(P < 0.05)，软骨组织p-PI3K、p-AKT、p-P65、NLRP3、GSDMD-N、Cleaved-Caspase1、白细胞介素18、白细胞介素1β蛋白表达显著降低(P < 0.01)。结果表明，右归丸可改善膝骨关节炎大鼠软骨细胞炎症反应，作用机制可能与抑制PI3K/AKT/NF-κB通路的激活，从而调控NLRP3/Caspase1/GSDMD通路介导的细胞焦亡相关。
https://orcid.org/0009-0004-8288-1792 (范元赫) 


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: 右归丸, 膝骨关节炎, 中药, 软骨, 炎症, 焦亡, 信号通路

Abstract: BACKGROUND: Yougui Pill is derived from Jingyue Quanshu. Studies have confirmed that Yougui Pill is highly effective in treating patients with knee osteoarthritis, but its mechanism of action remains unclear.
OBJECTIVE: To explore the potential molecular mechanism of Yougui Pill in improving knee osteoarthritis in rats. 
METHODS: Forty-eight SPF-grade Sprague-Dawley rats were randomly divided into four groups: blank control group, model group, Yougui Pill group and celecoxib group. Modified Hulth modeling method was used to establish the left knee joint model in the latter three groups. After wound healing, the rats were urged to move for 8 weeks. After modeling, the celecoxib group was given celecoxib suspension by gavage, the Yougui Pill group was given Yougui Pill decoction by gavage, and the blank control group and the model group were given the same volume of normal saline by gavage, once a day for 4 continuous weeks. The pathological changes of rat cartilage tissue were observed by hematoxylin-eosin staining, toluidine blue staining, and safranin O-fast green staining, and the ultrastructure of rat chondrocytes was observed by transmission electron microscope. ELISA was used to detect the levels of interleukin-18, interleukin-1β, and tumor necrosis factor-α in serum. The protein expressions of PI3K, AKT, NF-κB, p-PI3K, p-AKT, p-P65, NLRP3, GSDMD, GSDMD-N, Caspase1, Cleaved-Caspase1, interleukin-18, and interleukin-1β in cartilage tissues of rats were detected by western blot. 
RESULTS AND CONCLUSION: (1) Compared with the blank control group, the rats in the model group had severe cartilage edge destruction, cartilage tissue defect, thinning and disordered arrangement of cartilage layer cells, subchondral bone hyperplasia, disordered tide line, severe structural damage of chondrocytes and formation of pyroptosis. The levels of tumor necrosis factor-α, interleukin-18 and interleukin-1β in serum were significantly increased (P < 0.05). The protein expressions of p-PI3K, p-Akt, p-p65, NLRP3, GSDMD-N, Cleaved Caspase1, interleukin-18, and interleukin-1β in cartilage tissue were significantly increased (P < 0.01). (2) Compared with the model group, the Yougui Pill group and celecoxib group had normal cartilage structure, darker cartilage color, thicker cartilage, more complete chondrocyte membrane, and significantly reduced serum levels of tumor necrosis factor-α, interleukin-18 and interleukin-1β (P < 0.05). The protein expressions of p-PI3K, p-Akt, p-p65, NLRP3, GSDMD-N, Cleaved Caspase1, interleukin-18, and interleukin-1β in cartilage tissue were significantly decreased (P < 0.01). (3) Compared with the celecoxib group, the chondrocytes in the Yougui Pill group were arranged more regular, the articular cartilage surface was relatively smooth, the cartilage layer was thickened, the tide line was relatively complete, and the cell membrane of chondrocytes was relatively complete. The levels of tumor necrosis factor-α, interleukin-18 and interleukin-1β in serum were significantly decreased (P < 0.05). The protein expressions of p-PI3K, p-Akt, p-p65, NLRP3, GSDMD-N, Cleaved Caspase1, interleukin-18, and interleukin-1β in cartilage tissue were significantly decreased (P < 0.01). In conclusion, Yougui Pill can improve the inflammatory response of chondrocytes in rats with knee osteoarthritis, and its mechanism may be related to the inhibition of PI3K/AKT/NF-kB pathway activation, thereby regulating pyroptosis mediated via the NLRP3/Caspase-1/GSDMD pathway.

Key words: Yougui Pill, knee osteoarthritis, traditional Chinese medicine, cartilage, inflammation, pyroptosis, signaling pathway

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