实用医学杂志 ›› 2023, Vol. 39 ›› Issue (12): 1473-1479.doi: 10.3969/j.issn.1006⁃5725.2023.12.003

• 基础研究 • 上一篇    下一篇

电针对老年大鼠关节软骨及软骨下骨极化相关蛋白表达的影响

黄夏荣1 周君1 孙光华1 彭昕珂1 廖源2 刘静1 罗敷1 钟培瑞1 彭婷1 胡莉芝3    

  1. 南华大学衡阳医学院附属第一医院1 康复医学科、康复医学中心、康复医学实验室,3 肛肠科(湖南衡阳 421001);2 广州医科大学附属第六医院,清远市人民医院康复科(广东清远511500)

  • 出版日期:2023-06-25 发布日期:2023-06-25
  • 通讯作者: 胡莉芝 E⁃mail:1471485472@qq.com
  • 基金资助:
    国家自然科学基金项目(编号:81973917,82105022);湖南省自然科学基金面上项目(编号:2021JJ30616);湖南省自然科 学基金青年项目(编号:2021JJ40493);南华大学校级重点项目(编号:USCKF201902K02);湖南省卫生健康委项目(编号:20200464)

Effect of electroacupuncture on articular cartilage and subchondral polarization⁃related protein expression in aged rats

HUANG Xiarong*,ZHOU Jun,SUN Guanghua,PENG Xinke,LIAO Yuan,LIU Jing,LUO Fu, ZHONG Peirui,PENG Ting,HU Lizhi.   

  1. Department of Rehabilitation,Rehabilitation Medicine Center,Rehabilita⁃ tion Laboratory,the First Affiliated Hospital,Hengyang Medical School,University of South China,Hengyang 421001,China

  • Online:2023-06-25 Published:2023-06-25
  • Contact: HU Lizhi E⁃mail:1471485472@qq.com

摘要:

目的 探讨电针治疗对老年大鼠的关节软骨及软骨下骨极化相关蛋白表达的影响。 方法 16 只 24 月龄 SD 雄性大鼠随机分为老年组和电针组,每组 8 只;8 只 6 月龄 SD 雄性大鼠为青年组。 电针组接受电针治疗,穴取双侧“太溪”、“足三里”、“阳陵泉”、“血海”。电针参数:疏密波,疏波频率 3 Hz, 密波 15 Hz,电流强度 1 mA,1 次/d,30 min/次,每周 5 d,连续 8 周。其余两组用胶带固定无菌针,连接电针 仪但不开机。8 周后取材,ELISA 法检测血清Ⅱ型胶原 C 端肽(CTX⁃Ⅱ)含量;显微 CT(Micro⁃CT)检测左侧 胫骨软骨下骨微结构;番红⁃固绿染色在光镜下观察左胫骨平台软骨组织形态结构,并采用改良 Mankin′s 评分评估关节软骨退变程度。RT⁃qPCR、Western blot 检测基质金属蛋白酶⁃1(MMP⁃1),基质金属蛋白酶⁃3 (MMP⁃3),基质金属蛋白酶⁃13(MMP⁃13),肿瘤坏死因子⁃α(TNF⁃α),白细胞介素⁃1β(IL⁃1β),巨噬细胞甘 露糖受体(MMR),精氨酸酶 1(Arg1)mRNA 及蛋白表达水平。结果 (1)与青年组大鼠相比,老年组大鼠 血液中 CTX⁃Ⅱ含量增高(P < 0.05);番红⁃固绿染色显示大鼠软骨表面结构破坏严重,软骨与软骨下骨分 解不清,钙化严重,出现多重潮线,细胞数量明显减少,出现裂层,细胞形态结构异常,染色不均;改良 Mankin′s 评分增加(P < 0.05);骨密度、骨体积分数、骨小梁数量均降低(P < 0.05),骨小梁厚度,骨小梁间 距均增大(P < 0.05);MMP⁃1,MMP⁃3,MMP⁃13 mRNA 及蛋白表达升高(P < 0.05),TNF⁃α,IL⁃1β mRNA 及蛋 白表达升高(P < 0.05),MMR,Arg1 mRNA 及蛋白表达降低(P < 0.05)。(2)与老年组大鼠相比,电针组大鼠 的 CTX⁃Ⅱ含量降低(P < 0.05);番红-固绿染色显示大鼠软骨表面稍平整,未见明显裂层,软骨细胞数量较 老年组有所增加,染色较均匀;改良 Mankin′s 评分降低(P < 0.05);骨密度,骨体积分数,骨小梁数量均增 加(P < 0.05),骨小梁间距降低(P < 0.05),骨小梁厚度有降低趋势(P > 0.05);MMP⁃1,MMP⁃3,MMP⁃13 mRNA 及蛋白表达降低(P < 0.05),TNF⁃α,IL⁃1β mRNA 及蛋白表达降低(P < 0.05),MMR,Arg1 mRNA 及 蛋白升高(P < 0.05)。结论 电针可能通过促进老年大鼠 OA 关节软骨及软骨下骨 M2 极化,缓解关节软 骨降解及抑制软骨下骨骨质疏松,从而达到保护关节的作用。

关键词:

电针, 软骨, 软骨下骨, 极化, 骨关节炎

Abstract:

Objective To investigate the effect of electroacupuncture on articular cartilage and subchon⁃ dral polarization ⁃ related protein expression in aged rats. Methods Sixteen 24⁃month ⁃old SD male rats were randomly divided into old group and electroacupuncture group,with 8 rats in each group. Eight SD male rats aged 6 months were youth group. Electroacupuncture group received electroacupuncture treatment,and the two sides of the“Taixi”,“Zusanli”,“Yanglingquan”,“xuehai”were selected. Electroacupuncture parameters:density wave, sparse wave frequency 3 Hz,dense wave 15 Hz,current intensity 1 mA,once a day,30 minutes each time,5 days a week,consecutive 8 weeks. The other two groups fixed sterile needles with adhesive tape,connected the elec⁃ troacupuncture instrument but did not turn on. After 8 weeks,the content of C⁃terminal peptide(CTX⁃Ⅱ)in serum of type Ⅱ collagen was determined by ELISA. Microscopic CT(Micro⁃CT)was used to detect the microstruc⁃ ture of the left tibia subchondral bone. The left tibial plateau cartilage was observed under light microscope with saffron⁃solid green staining,and the degree of articular cartilage degeneration was evaluated by modified Mankin′s score. RT⁃qPCR and Westernblot were used to detect matrix metalloproteinase 1(MMP⁃1),matrix metallopro⁃ teinase 3(MMP⁃3),matrix metalloproteinase 13(MMP⁃13),tumor necrosis factor α(TNF⁃α),interleukin 1β (IL⁃1β),and macrophage mannose⁃receptor(MMR). Expression levels of arginase 1(Arg1)mRNA and protein. Results (1)Compared with the young group,the CTX⁃Ⅱin the blood of the old group increased(P < 0.05),Saf⁃ fron O⁃fast green staining showed that the structure of rat cartilage was seriously damaged,cartilage and subchon⁃ dral bone decomposed indistinctly,calcification was severe,multiple tidal lines appeared,the number of cells decreased significantly,split layer appeared,cell morphology and structure was abnormal,and staining was uneven. and the modified Mankin′s score increased(P < 0.05). Bone mineral density,bone volume fraction and number of bone trabeculae decreased(P < 0.05),trabecular thickness and trabecular spacing increased(P < 0.05),MMP1, MMP3,MMP13 mRNA and protein expression increased(P < 0.05),TNF⁃α,IL⁃1 β mRNA and protein expres⁃ sion increased(P < 0.05),MMR,Arg1mRNA and protein decreased(P < 0.05).(2)Compared with the senile group,the CTX⁃Ⅱdecreased in the electroacupuncture group(P < 0.05),The saffron O⁃fast green staining showed that the cartilage of the rats was slightly smooth and there was no obvious cleavage layer. The number of chondro⁃ cytes in the aged group was higher than that in the aged group,and the staining was more uniform;and the modi⁃ fied Mankin′s score was decreased(P < 0.05). Bone mineral density,bone volume fraction and trabecular number increased(P < 0.05),trabecular spacing decreased(P < 0.05),trabecular thickness tended to decrease(P > 0.05),MMP1,MMP3,MMP13mRNA and protein expression decreased(P < 0.05),TNF⁃α,IL⁃1 β mRNA and protein expression decreased(P < 0.05),MMR,Arg1mRNA and protein increased(P < 0.05). Conclusion Electroacupuncture may protect joints by promoting OA articular cartilage and subchondral bone M2 polarization, alleviating articular cartilage degradation and inhibiting subchondral bone osteoporosis in aged rats.

Key words:

electroacupuncture, cartilage, subchondral bone, polarization, osteoarthritis