中国组织工程研究 ›› 2023, Vol. 27 ›› Issue (20): 3202-3208.doi: 10.12307/2023.429

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

温针灸抑制NLRP3炎症小体激活改善兔膝骨关节炎的软骨损伤

武永利1,2,李  龙3,刘君伟1,刘  娣1,王  铎3   

  1. 1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,2少数民族医药现代化教育部重点实验室,3中医学院,宁夏回族自治区银川市  750004
  • 收稿日期:2022-04-16 接受日期:2022-06-23 出版日期:2023-07-18 发布日期:2022-11-19
  • 通讯作者: 武永利,教授,主任医师,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:武永利,男,1968 年生,宁夏回族自治区银川市人,汉族,教授,主任医师,博士生导师,主要从事退行性脊柱关节病的基础及临床研究。
  • 基金资助:
    国家自然科学基金项目(82160939,81760891),项目负责人:武永利;宁夏回族自治区第二批科技领军人才项目(KJT201610),项目负责人:武永利

Warm-needling moxibusion inhibits NLRP3 inflammasome activation and improves cartilage injury in a rabbit model of knee osteoarthritis

Wu Yongli1, 2, Li Long3, Liu Junwei1, Liu Di1, Wang Duo3   

  1. 1Department of Orthopedics and Traumatology of Traditional Chinese Medicine, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 2Key Laboratory of Ningxia Ethnomedicine Modernization, Ministry of Education, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 3School of Traditional Chinese Medicine, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Received:2022-04-16 Accepted:2022-06-23 Online:2023-07-18 Published:2022-11-19
  • Contact: Wu Yongli, Department of Orthopedics and Traumatology of Traditional Chinese Medicine, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Key Laboratory of Ningxia Ethnomedicine Modernization, Ministry of Education, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • About author:Wu Yongli, Professor, Chief physician, Doctoral supervisor, Department of Orthopedics and Traumatology of Traditional Chinese Medicine, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Key Laboratory of Ningxia Ethnomedicine Modernization, Ministry of Education, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Supported by:
    the National Natural Science Foundation of China, Nos. 82160939 and 81760891 (both to WYL); The Second Batch of Science and Technology Leading Talents Project of Ningxia Hui Autonomous Region, No. KJT201610 (to WYL)

摘要:


文题释义:

温针灸:是针刺效应与灸热效应相结合的一种中医特色外治法,是在针刺得气后将艾绒搓团捻裹于针柄上点燃,借助针体将热力传入穴位。针刺穴位局部产生的酸胀重麻的针感配以艾灸的温热效应,更有利于得气产生及经络感传。该法具有温经通络、消肿止痛的功效,可改善病变部位的气血运行,起到抗炎、消肿、镇痛的作用。该疗法简便验廉、疗效确切,被广泛应用于治疗膝骨关节炎。
NLRP3炎症小体:是由胞浆内模式识别受体参与组装的多蛋白复合物,是天然免疫系统的重要组成部分,其在接收机体损伤信号及释放相关炎性因子的过程中起关键作用,又被称炎性反应的感受器与调节器。炎症小体的活化能够诱导产生多种炎性因子,进而加重炎性反应。NLRP3炎症小体在膝骨关节炎的发生发展过程中发挥着关键作用,因此作为炎症反应的核心,NLRP3炎症小体可能为膝骨关节炎的治疗提供新的靶点。

背景:温针灸治疗膝骨关节炎疗效显著,但其作用机制尚不明确。
目的:观察温针灸对兔膝骨关节炎软骨组织中NLRP3、基质金属蛋白酶3、基质金属蛋白酶13、Ⅰ型胶原蛋白表达的影响。
方法:取6月龄新西兰雄兔40只,采用随机数字表法分成空白组、模型组、温针灸组、抑制剂组,每组10只:除空白组外,其他3组均采用石膏管型固定法建立右后肢膝骨关节炎模型。造模成功后,温针灸组进行温针灸治疗,1次/d,连续治疗4周;抑制剂组膝关节腔内注射NLRP3抑制剂MCC950,1次/周,注射4次。干预结束后,采用Lequesne MG评分评估兔行为学改变,苏木精-伊红染色观察软骨形态学结构,免疫组织化学染色及蛋白质印迹法检测软骨组织中NLRP3、基质金属蛋白酶3、基质金属蛋白酶13、Ⅰ型胶原蛋白的表达。
结果与结论:①Lequesne MG评分:模型组Lequesne MG评分高于空白组(P < 0.05),温针灸组、抑制剂组Lequesne MG评分低于模型组(P < 0.05);②苏木精-伊红染色:与空白组比较,模型组软骨细胞数量减少,结构完整性被破坏,Mankin’s评分升高(P < 0.05);与模型组比较,温针灸组、抑制剂组软骨细胞分布较均匀,软骨层结构相对清晰,Mankin’s评分降低(P < 0.05);③免疫组织化学与蛋白质印迹检测:与空白组比较,模型组NLRP3、基质金属蛋白酶3、基质金属蛋白酶13表达升高(P < 0.05),Ⅰ型胶原蛋白表达降低(P < 0.05);与模型组比较,温针灸组与抑制剂组NLRP3、基质金属蛋白酶3、基质金属蛋白酶13表达降低(P < 0.05),Ⅰ型胶原蛋白表达升高(P < 0.05);④结果表明:温针灸可抑制NLRP3炎性启动因子的激活、下调炎性因子基质金属蛋白酶3,13的释放,降低炎性放大反应,减缓Ⅰ型胶原蛋白的降解,降低膝骨关节炎的关节软骨损伤及局部破坏。
https://orcid.org/0000-0001-9774-453X(武永利)

关键词: 温针灸, 膝骨关节炎, NLRP3炎症小体, 软骨损伤, 基质金属蛋白酶

Abstract: BACKGROUND: Warm-needling moxibusion has a significant clinical effect on knee osteoarthritis, but its therapeutic mechanism remains unclear. 
OBJECTIVE: To observe the effect of warm-needling moxibusion on the expression of NLRP3, matrix metalloproteinases 3, 13, and type I collagen in cartilage tissue of rabbit knee osteoarthritis.
METHODS: Forty 6-month-old male New Zealand rabbits were randomly divided into blank group, model group, warm-needling moxibusion group, and NLRP3 inhibitor group, with 10 rabbits in each group. Except for the blank group, osteoarthritic models of the right hind limb were established in the other three groups using plaster cast fixation. After modeling, intervention with warm-needling moxibusion in the warm-needling moxibusion group was performed once a day and lasted for 4 weeks; in the NLRP3 inhibitor group, MCC950, an NLRP3 inhibitor, was injected into the knee joint cavity, once a week, for four times in total. Lequesne MG score was used for behavioral evaluation, hematoxylin-eosin staining was used to observe the morphological structure of cartilage tissue, and the expression of NLRP3, matrix metalloproteinases 3, 13, and type I collagen in chondrocytes was detected by immunohistochemistry and western blot.
RESULTS AND CONCLUSION: Lequesne MG score: Compared with the blank group, Lequesne MG score increased in the other three groups after modeling (P < 0.05). After treatment, the Lequesne MG scores were significantly decreased in the warm-needling moxibusion group and NLRP3 inhibitor group compared with the model group (P < 0.05). Hematoxylin-eosin staining: compared with the blank group, the number of chondrocytes was decreased, the structural integrity was destroyed, and the Mankin’s score was significantly increased in the model group (P < 0.05). Compared with the model group, the distribution of chondrocytes was more uniform, the structure of cartilage layer was relatively clear, and the Mankin’s score was decreased in the warm-needling moxibusion group and NLRP3 inhibitor group (P < 0.05). Immunohistochemistry and western blot detection: compared with the blank group, the expression of NLRP3, matrix metalloproteinases 3 and 13 was significantly increased (all P < 0.05), and the expression of type I collagen was significantly down-regulated in the model group (P < 0.05). Compared with the model group, the expression of NLRP3, matrix metalloproteinases 3 and 13 were significantly decreased in the warm-needling moxibusion group and NLRP3inhibitor group (all P < 0.05), and the expression of type I collagen was significantly up-regulated (P < 0.05). To conclude, warm-needling moxibusion can inhibit the activation of NLRP3 inflammatory promoter, down-regulate the release of inflammatory factors, matrix metalloproteinases 3 and 13, reduce the inflammatory amplification reaction, and delay the degradation of type I collagen, thereby reducing cartilage injury and local damage of articular cartilage due to knee osteoarthritis.

Key words: warm-needling moxibusion, knee osteoarthritis, NLRP3 inflammasome, cartilage injury, matrix metalloproteinase

中图分类号: