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经皮耳穴迷走神经刺激对类风湿关节炎大鼠骨与软骨破坏的影响 被引量:7

Effects of transcutaneous auricular vagus nerve stimulation on bone and cartilage destruction in rats with rheumatoid arthritis
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摘要 目的:观察经皮耳穴迷走神经刺激(taVNS)对胶原诱导型关节炎(CIA)大鼠关节骨和软骨破坏的影响,探讨taVNS缓解类风湿关节炎效应的细胞与分子机制。方法:SD大鼠随机分为正常组、模型组与taVNS组,每组各12只。通过在大鼠尾根部多点注射牛Ⅱ型胶原和弗氏非完全佐剂混合而成的乳剂制备CIA模型。taVNS组大鼠予以双侧耳甲区穴位经皮电刺激干预,每次30 min,1次/d,连续28 d。观测并评估大鼠关节炎指数(AI);通过番红O-固绿染色法观察大鼠踝关节的软骨损毁情况;TRAP染色法检测大鼠关节组织中的破骨细胞生成情况;X线和Micro-CT影像学法观察大鼠的骨侵蚀表现,并计算骨表面积/骨体积(BS/BV);Western blot法检测大鼠滑膜组织中基质金属蛋白酶(MMP)-1、MMP-3、MMP-13和核因子κB受体激活因子配体(RANKL)及骨保护素(OPG)的蛋白表达。结果:与正常组比较,CIA大鼠呈现出典型的类风湿关节炎症状,AI显著升高(P<0.05),taVNS组大鼠的AI较模型组显著降低(P<0.05)。与正常组比较,模型组大鼠踝关节软骨基质丢失,软骨层变簿,软骨明显损毁,关节内破骨细胞数量显著增加(P<0.01);影像学结果显示踝关节骨质丢失、骨的三维结构破坏,BS/BV显著升高(P<0.01),骨侵蚀明显加重;踝关节滑膜组织中MMP-1、MMP-3和MMP-13表达量及RANKL/OPG显著升高(P<0.01,P<0.05),OPG蛋白表达明显降低(P<0.05)。与模型组比较,taVNS组大鼠踝关节软骨层较完整,软骨损毁有所缓解;关节中破骨细胞数量显著减少(P<0.01);BS/BV明显降低(P<0.01),骨质丢失及骨结构破坏等骨侵蚀症状明显缓解;踝关节滑膜中MMP-1、MMP-3、MMP-13的表达水平及RANKL/OPG显著降低(P<0.05,P<0.01),OPG蛋白表达显著升高(P<0.05)。结论:taVNS可有效缓解CIA大鼠的骨与软骨破坏症状,其机制可能与taVNS减少了关节组织中破骨细胞的数量,降低滑膜内MMP-1、MMP-3、MMP-13表达水平及RANKL/OPG有关。 Objective To observe the alleviating effect of transcutaneous auricular vagus nerve stimulation(taVNS)on articular cartilage and bone destruction in rats with collagen-induced arthritis(CIA),and explore the cellular and molecular mechanisms of taVNS against rheumatoid arthritis(RA).Methods The male SD rats were randomly divided into normal control group(n=12),model group(n=12),and taVNS group(n=12).The CIA rat model was established by multi-point injection of emulsion prepared from typeⅡbovine collagen and Freund’s incomplete adjuvant into the root of rat tail.The rats in the taVNS group were treated with taVNS at bilateral auricular conchae,30 min per time,once a day,for consecutive 28 d.The cartilage destruction of the ankle joint was observed by safranin O-fast green staining,the production of osteoclasts in the joint tissue by tartrate-resistant acid phosphatase(TRAP)staining,and the bone erosion by X-ray and Micro-CT imaging.The protein expression levels of matrix metalloproteinase(MMP)-1,MMP-3,MMP-13,receptor activator of nuclear factor-κB ligand(RANKL),and osteoprotegerin(OPG)in the synovial tissues were detected by Western blot.Results Compared with the normal control group,the CIA rats presented with typical RA symptoms and elevated arthritis index(AI,P<0.05).After intervention with taVNS,the AI remarkably declined in comparison with that in the model group(P<0.05).Compared with the control group,the model group displayed loss of cartilage matrix in the ankle joint,thinned cartilage layer,obvious cartilage damage,and increased number of osteo-clasts in the joint(P<0.01);the imaging results showed bone loss and three-dimensional structural destruction of ankle joint and aggravated bone erosion(P<0.01);the expression levels of MMP-1,MMP-3 and MMP-13,and RANKL/OPG ratio were significantly elevated in the synovial tissue of ankle joint(P<0.01,P<0.05),while the expression level of OPG was decreased(P<0.05).Compared with the model group,taVNS resulted in relatively intact cartilage layer of ankle joint,alleviated cartilage destruction,decreased number of osteoclasts(P<0.01),improved bone erosion,loss,and three-dimensional structural destruction(P<0.01),and diminished MMP-1,MMP-3,and MMP-13 expression and RANKL/OPG ratio in the synovial tissue of ankle joint(P<0.05,P<0.01),while the expression level of OPG was increased(P<0.05).Conclusion taVNS effectively relieves bone and cartilage destruction in CIA rats,which might be related to its efficacy in reducing the production of osteoclasts in joint tissues and down-regulating the expression levels of MMP-1,MMP-3 and MMP-13,and RANKL/OPG ratio.
作者 阎琪琪 孙诗月 谭连红 史亦南 乔丽娜 杨永升 YAN Qi-qi;SUN Shi-yue;TAN Lian-hong;SHI Yi-nan;QIAO Li-na;YANG Yong-sheng(Institute of Acu-moxibustion,China Academy of Chinese Medical Sciences,Beijing 100700,China)
出处 《针刺研究》 CAS CSCD 北大核心 2022年第3期237-243,共7页 Acupuncture Research
基金 北京市自然科学基金面上项目(No.7202140) 中央级公益性科研院所基本科研业务费专项资金资助(No.ZZ11-097、ZZ16011、ZZ-ZR2017007、ZZ13-YQ-063)。
关键词 经皮耳穴迷走神经刺激 骨与软骨 类风湿关节炎 Transcutaneous auricular vagus nerve stimulation(taVNS) Bone and cartilage Rheumatoid arthritis
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