中国组织工程研究

中国组织工程研究 ›› 2026, Vol. 30 ›› Issue (36): 9445-9452.doi: 10.12307/2026.898

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

温针灸抑制细胞焦亡改善兔膝关节软骨退变

刘  娣1，王明磊2，李  春1，王  铎3，李  龙1，武永利1，4   

  1. 宁夏医科大学总医院，1中医骨伤科，2放射科，宁夏回族自治区银川市  750004；3灵武市中医院，宁夏回族自治区灵武市  751600；4宁夏医科大学宁夏少数民族医药现代化教育部重点实验室，宁夏回族自治区银川市  750004
  • 收稿日期:2025-09-29 修回日期:2026-03-04 出版日期:2026-12-28 发布日期:2026-05-21
  • 通讯作者: 武永利，硕士，主任医师，教授，博士生导师，宁夏医科大学总医院中医骨伤科，宁夏回族自治区银川市 750004；宁夏医科大学宁夏少数民族医药现代化教育部重点实验室，宁夏回族自治区银川市 750004
  • 作者简介:刘娣，女，1989年生，宁夏回族自治区银川市人，汉族，博士，副主任医师，主要从事退行性脊柱关节病的临床与基础研究。
  • 基金资助:
    国家自然科学基金资助项目(82160939)，项目负责人：武永利；宁夏自然科学基金项目(2023AAC03538)，项目负责人：刘娣；宁夏自然科学基金项目(2022AAC03585)，项目负责人：王明磊；宁夏教育厅项目(NYG20241)，项目负责人：刘娣

Warm acupuncture inhibits pyroptosis and improves cartilage degeneration in rabbit knee joints

Liu Di1, Wang Minglei2, Li Chun1, Wang Duo3, Li Long1, Wu Yongli1, 4   

  1. 1Department of Traditional Chinese Medicine Orthopedics and Traumatology, 2Department of Radiology, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 3Lingwu Municipal Hospital of Traditional Chinese Medicine, Lingwu 751600, Ningxia Hui Autonomous Region, China; 4Key Laboratory of Ningxia Minority Medicine Modernization, Ministry of Education, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Received:2025-09-29 Revised:2026-03-04 Online:2026-12-28 Published:2026-05-21
  • Contact: Wu Yongli, MS, Chief physician, Professor, Doctoral supervisor, Department of Traditional Chinese Medicine Orthopedics and Traumatology, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Key Laboratory of Ningxia Minority Medicine Modernization, Ministry of Education, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • About author:Liu Di, PhD, Associate chief physician, Department of Traditional Chinese Medicine Orthopedics and Traumatology, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Supported by:
    National Natural Science Foundation of China, No. 82160939 (to WYL); Natural Science Foundation of Ningxia, Nos. 2023AAC03538 (to LD) and 2022AAC03585 (to WML); Project of the Department of Education of Ningxia, No. NYG20241 (to LD)

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摘要：



文题释义：
温针灸疗法：是一种将针刺与艾灸相结合的中医外治法之一，通过将刺入穴位的毫针针柄上安装艾炷并点燃，使热力通过针体传导至穴位深处，通过“热力-机械”的复合刺激发挥抗炎、镇痛、改善循环等效应，起到温经通痹的作用。
细胞焦亡：是一种伴随持续性炎症反应的细胞死亡方式，主要依赖于Caspase-1的介导，活化后的Caspase-1可触发并分解消皮素D产生N端，从而启动细胞焦亡，导致病变局部促炎因子的升高，触发并放大炎症级联反应，而大量炎性因子释放又会加剧细胞焦亡，引起“炎症-焦亡”恶性循环，造成组织严重损伤。细胞焦亡驱动下的持续性炎症环境是导致膝骨关节炎软骨进行性破坏的重要原因。
Toll样受体4/核因子κB信号通路：是经典的炎性反应信号通路之一，也是介导细胞焦亡发生的重要信号通路。Toll样受体4作为模式识别受体，能够快速识别内源性或外源性危险信号，进而激活核因子κB信号通路，活化后的核因子κB能够显著上调炎症小体组分(如
pro-caspase-1)，并调控下游焦亡关键执行蛋白消皮素D的表达，诱发细胞焦亡发生，进一步促进促炎细胞因子白细胞介素1β、白细胞介素18的产生。

背景：温针灸疗法治疗膝骨关节炎的疗效显著，可通过抑制关节周围炎症改善关节软骨退变，但该作用是否与Toll样受体4/核因子κB信号通路介导的细胞焦亡相关尚不清楚。
目的：从细胞焦亡角度探讨温针灸治疗膝骨关节炎的作用机制。
方法：采用随机数字表法将40只新西兰兔随机分为空白组、模型组、西药组及温针灸组，每组10只。空白组不进行任何干预，其余3组采用石膏管型固定法制备膝骨关节炎模型；造模4周后，模型组兔置于固定器上固定15 min，每天1次，连续固定2周；温针灸组选取“鹤顶”“内膝眼”及“外膝眼”进行温针灸治疗，每次15 min，每天1次，连续治疗2周；西药组灌胃给予塞来昔布，每天1次，连续给药2周。治疗结束后第2天取材，ELISA检测关节液中白细胞介素1β、白细胞介素18水平，苏木精-伊红染色观察膝关节软骨形态学改变，qPCR、免疫组化、Western blot检测膝关节软骨组织中Toll样受体4、核因子κB p65、Caspase-1及消皮素D表达。
结果与结论：①模型组白细胞介素1β、白细胞介素18水平高于空白组(P < 0.05)，温针灸组和西药组白细胞介素1β、白细胞介素18水平低于模型组(P < 0.05)；②苏木精-伊红染色显示，模型组软骨表面粗糙不平，软骨受损明显，软骨细胞分布不规则、排列紊乱；温针灸组和西药组软骨层表面较为光滑，软骨细胞分布比较均匀整齐；③qPCR、免疫组化、Western blot检测显示，与空白组比较，模型组软骨组织中Toll样受体4、核因子κB p65、Caspase-1及消皮素D的mRNA和蛋白表达均升高(P < 0.05)；与模型组比较，温针灸组和西药组软骨组织中Toll样受体4、核因子κB p65、Caspase-1及消皮素D的mRNA和蛋白表达均降低(P < 0.05)；④结果表明，温针灸可通过调控Toll样受体4/核因子κB信号通路下调焦亡相关蛋白Caspase-1、消皮素D及下游炎性因子白细胞介素1β、白细胞介素18 的表达，抑制细胞焦亡，进而改善兔膝骨关节炎的关节软骨退变。
https://orcid.org/0000-0002-2209-8219(刘娣)


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: 温针灸, 膝骨关节炎, 细胞焦亡, Toll样受体4/核因子κB信号通路, Caspase-1, 消皮素D

Abstract: BACKGROUND: Warm acupuncture has a significant therapeutic effect on knee osteoarthritis. It can improve articular cartilage degeneration by inhibiting periarticular inflammation. However, whether this effect is related to pyroptosis mediated by the Toll-like receptor 4/nuclear factor-κB (TLR4/NF-κB) signaling pathway remains unclear.
OBJECTIVE: To investigate the mechanism of warm acupuncture in the treatment of knee osteoarthritis from the perspective of pyroptosis.
METHODS: Forty New Zealand rabbits were randomly divided into blank, model, western medicine, and warm acupuncture groups using a random number table, with 10 rabbits in each group. The blank group received no intervention, while knee osteoarthritis models were established in the other three groups using the plaster cast fixation method. Four weeks after modeling, the model group was immobilized in a fixator for 15 minutes once daily for 2 consecutive weeks. The warm acupuncture group received warm acupuncture at “Heding” (EX-LE2), “Neixiyan” (EX-LE4), and “Waixiyan” (ST35) for 15 minutes once daily for 2 weeks. The western medicine group received celecoxib by gavage once daily for 2 weeks. Samples were collected on the second day after treatment. Interleukin-1β and interleukin-18 levels in synovial fluid were detected by ELISA. Morphological changes in knee cartilage were observed by hematoxylin-eosin staining. The expression of TLR4, NF-κB p65, Caspase-1, and gasdermin D in knee cartilage was detected by qPCR, immunohistochemistry, and western blot.
RESULTS AND CONCLUSION: (1) Interleukin-1β and interleukin-18 levels were significantly higher in the model group than in the blank group (P < 0.05), and significantly lower in the warm acupuncture and western medicine groups than in the model group (P < 0.05). (2) Hematoxylin-eosin staining results showed that the cartilage surface in the model group was rough and significantly damaged, with irregular and disorganized chondrocyte distribution. In the warm acupuncture and western medicine groups, the cartilage surface was relatively smooth, and chondrocytes were more uniformly and regularly distributed. (3) Results from qPCR, immunohistochemistry, and western blot showed that compared with the blank group, the mRNA and protein expression levels of TLR4, NF-κB p65, Caspase-1, and gasdermin D were significantly increased in the model group (P < 0.05). Compared with the model group, the mRNA and protein expression levels of TLR4, NF-κB p65, Caspase-1, and gasdermin D were significantly decreased in the warm acupuncture and western medicine groups (P < 0.05). These findings indicate that warm acupuncture can alleviate articular cartilage degeneration in rabbits with knee osteoarthritis by regulating the TLR4/NF-κB signaling pathway to downregulate the expression of pyroptosis-related proteins Caspase-1 and gasdermin D, as well as downstream inflammatory factors interleukin-1β and interleukin-18, thereby inhibiting pyroptosis.

Key words: warm acupuncture, knee osteoarthritis, pyroptosis, Toll-like receptor 4/nuclear factor-κB signaling pathway, Caspase-1, gasdermin D

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