中国组织工程研究

中国组织工程研究 ›› 2024, Vol. 28 ›› Issue (8): 1174-1179.doi: 10.12307/2023.955

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

电针干预老年膝骨关节炎大鼠关节软骨及软骨下骨P53、P21的表达

黄夏荣1，2，3，胡莉芝4，孙光华1，2，3，彭昕珂1，2，3，廖  瑛1，2，3，廖  源5，刘  静1，2，3，尹林伟1，2，3，钟培瑞1，2，3，彭  婷1，2，3，周  君1，2，3，6，屈萌艰1，2，3
  

  1. 南华大学衡阳医学院附属第一医院，1康复医学科，2康复医学中心，3康复医学实验室，4肛肠科，湖南省衡阳市  421001；5广州医科大学附属第六医院，清远市人民医院康复科，广东省清远市  511500；6康复医学四川省重点实验室，四川省成都市  610041
  • 收稿日期:2022-10-28 接受日期:2022-12-24 出版日期:2024-03-18 发布日期:2023-07-18
  • 通讯作者: 屈萌艰，在读博士，主管技师，南华大学衡阳医学院附属第一医院康复医学科，康复医学中心，康复医学实验室，湖南省衡阳市 421001
  • 作者简介:黄夏荣，男，1989年生，江西省吉安市人，主管技师，主要从事骨关节炎和骨质疏松的临床与基础研究。
  • 基金资助:
    国家自然科学基金项目(81973917)，项目负责人：周君；国家自然科学基金项目(82105022)，项目负责人：廖源；湖南省自然科学基金面上项目(2021JJ30616)，项目负责人：廖源；湖南省自然科学基金青年项目(2021JJ40493)，项目负责人：黄夏荣；南华大学校级重点项目(USCKF201902K02)，项目负责人：廖瑛

Effect of electroacupuncture on the expression of P53 and P21 in articular cartilage and subchondral bone of aged rats with knee osteoarthritis

Huang Xiarong1, 2, 3, Hu Lizhi4, Sun Guanghua1, 2, 3, Peng Xinke1, 2, 3, Liao Ying1, 2, 3, Liao Yuan5, Liu Jing1, 2, 3, Yin Linwei1, 2, 3, Zhong Peirui1, 2, 3, Peng Ting1, 2, 3, Zhou Jun1, 2, 3, 6, Qu Mengjian1, 2, 3   

  1. 1Department of Rehabilitation, 2Rehabilitation Medicine Center, 3Rehabilitation Laboratory, 4Department of Proctology, the First Affiliated Hospital of Hengyang Medical School, University of South China, Hengyang 421001, Hunan Province, China; 5Department of Rehabilitation, Qingyuan People’s Hospital, the Sixth Affiliated Hospital of Guangzhou Medical University, Qingyuan 511500, China; 6Key Laboratory of Rehabilitation Medicine in Sichuan Province, Chengdu 610041, Sichuan Province, China
  • Received:2022-10-28 Accepted:2022-12-24 Online:2024-03-18 Published:2023-07-18
  • Contact: Qu Mengjian, MD candidate, Technologist-in-charge, Department of Rehabilitation, Rehabilitation Medicine Center, Rehabilitation Laboratory, the First Affiliated Hospital of Hengyang Medical School, University of South China, Hengyang 421001, Hunan Province, China
  • About author:Huang Xiarong, Technologist-in-charge, Department of Rehabilitation, Rehabilitation Medicine Center, Rehabilitation Laboratory, the First Affiliated Hospital of Hengyang Medical School, University of South China, Hengyang 421001, Hunan Province, China
  • Supported by:
    National Natural Science Foundation of China, Nos. 81973917 (to ZJ) and 82105022 (to LY); Natural Science Foundation of Hunan Province (General Program), No. 2021JJ30616 (to LY); Natural Science Foundation of Hunan Province (Youth Program), No. 2021JJ40493 (to HXR); University Key Project of South China University, No. USCKF201902K02 (to LY)

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摘要：


文题释义：

骨关节炎：是一种与年龄、性别、肥胖和关节损伤相关的异质性疾病，其病理表现主要为关节软骨的软化、纤维化、溃疡形成和缺失，软骨下骨硬化、骨赘形成及滑膜炎等。
关节软骨：是一种复杂的特殊结构，其覆盖在组成关节的骨端，功能是为关节在一定范围的活动提供一种低摩擦的接触表面，并可缓冲关节活动过程中的冲击力。


背景：膝骨关节炎的治疗方法有很多，其中电针作为一种重要的非药物治疗方法，治疗膝骨关节炎疗效确切，目前其确切的作用机制尚不明确。

目的：探讨电针对老年大鼠膝骨关节炎关节软骨及软骨下骨P53、P21表达的影响。
方法：青年组为8只6月龄SD雄性大鼠，16只24月龄SD雄性大鼠按随机抽样法分为老年组和电针组，每组8只。电针组大鼠接受电针刺激，每周5 d，1次/d，连续刺激8周，其余两组不做处理。8周后取材，ELISA法检测外周血Ⅱ型胶原C端肽水平，番红O-固绿染色对左膝关节软骨及软骨下骨进行形态学观察；改良Mankin’s评分评估膝关节软骨退变程度；显微CT检测左膝关节软骨及软骨下骨微结构；RT-qPCR、Western blot检测右膝关节软骨及软骨下骨基质金属蛋白酶13、P53、P21 mRNA及蛋白表达水平。

结果与结论：①与青年组相比，老年组大鼠外周血Ⅱ型胶原C端肽水平升高(P < 0.05)；显微CT显示老年组大鼠骨体积分数、骨密度、骨小梁数量均降低(P < 0.05)，骨小梁分离度增大(P < 0.05)；番红O-固绿染色显示老年组大鼠软骨表层不平整，出现裂层，软骨细胞形态不均，染色不均匀，软骨与软骨下骨分界模糊，基质损失严重；Mankin’s评分增加(P < 0.05)；基质金属蛋白酶13、P53、P21 mRNA表达升高(P < 0.05)，蛋白表达升高(P < 0.05)；②与老年组相比，电针组大鼠Ⅱ型胶原C端肽水平降低(P < 0.05)；显微CT显示电针组大鼠骨体积分数、骨密度、骨小梁数量均升高(P < 0.05)，骨小梁分离度减小(P < 0.05)；番红O-固绿染色显示软骨表面较平整，红染较均匀，细胞形态结构介于青年组和老年组之间；Mankin’s 评分降低(P < 0.05)；基质金属蛋白酶13、P21 mRNA表达降低(P < 0.05)，P53 mRNA表达有降低趋势，但差异无显著性意义(P > 0.05)；基质金属蛋白酶13、P53蛋白表达降低(P < 0.05)，P21蛋白表达有下降趋势，差异无统计学意义(P > 0.05)；③结果表明电针可能通过抑制P53和P21的表达而延缓老年大鼠关节软骨退变，并抑制关节软骨下骨骨质疏松，从而达到保护关节的作用延缓关节衰老。

https://orcid.org/0000-0001-9215-5744(黄夏荣)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 电针, 膝骨关节炎, 软骨, 软骨下骨, P53蛋白, P21蛋白

Abstract: BACKGROUND: There are many treatment methods for knee osteoarthritis, among which electroacupuncture, as an important non-drug treatment, is effective in the treatment of knee osteoarthritis, but its exact mechanism is not clear.
OBJECTIVE: Effect of electroacupuncture on the expression of p53 and P21 in articular cartilage and subchondral bone of aged rats with knee osteoarthritis.
METHODS: Eight 6-month-old male Sprague-Dawley rats were included in the young group and sixteen 24-month-old male Sprague-Dawley rats were randomly divided into old group (n=8) and electroacupuncture group (n=8). The rats in the electroacupuncture group received electroacupuncture stimulation once a day, 5 days a week, for 8 continuous weeks, and the other two groups did not do any treatment. Eight weeks later, the level of type II collagen C-terminal peptide in peripheral blood was detected by ELISA, the morphology of left knee cartilage and subchondral bone was observed by safranin O-fast green staining, the degree of knee cartilage degeneration was evaluated by modified Mankin’s score, the microstructure of left knee cartilage and subchondral bone was detected by micro-CT, and the expression levels of matrix metalloproteinase 13, P53, P21 mRNA and protein were detected by RT-PCR and western blot respectively.
RESULTS AND CONCLUSION: Compared with the young group, the level of C-terminal peptide of type II collagen in the peripheral blood was increased in the old group (P < 0.05). The micro-CT results showed that the bone volume fraction, bone mineral density and the number of bone trabeculae were decreased in the old group compared with the young group (P < 0.05), while the trabecular separation increased (P < 0.05). Safranin O-fast green staining showed that in the old group, the surface layer of cartilage was uneven with fissures, the morphology of chondrocytes was irregular and stained unevenly, the boundary between the cartilage and subchondral bone was blurred, and the matrix loss was serious. The Mankin’s score was higher in the old group than the young group (P < 0.05). The expression of matrix metalloproteinase 13, P53, P21 at mRNA and protein levels increased in the old group compared with the young group (P < 0.05). Compared with the old group, electroacupuncture decreased the level of C-terminal peptide of type II collagen (P < 0.05), increased the bone volume fraction, bone mineral density and the number of bone trabeculae (P < 0.05), and decreased the trabecular separation (P < 0.05). Safranin O-fast green staining showed that in the electroacupuncture group, the surface of cartilage was smooth and red staining was uniform, and the cell morphology and structure were between the young group and the old group. Following electroacupuncture treatment, the Mankin’s score (P < 0.05), matrix metalloproteinase 13 and P21 mRNA expression (P < 0.05), and matrix metalloproteinase 13 and P53 protein expression decreased (P < 0.05), while there was a decreasing trend of P53 mRNA and P21 protein expression, but with no statistical significance (P > 0.05). To conclude, electroacupuncture may delay articular cartilage degeneration and subchondral osteoporosis in aged rats by inhibiting the expression of P53 and P21, so as to protect joints and delay joint aging.

Key words: electroacupuncture, knee osteoarthritis, cartilage, subchondral bone, P53 protein, P21 protein

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