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游泳运动对膝骨关节炎小鼠软骨退变和NF-κB激活的影响

来源:用户上传      作者:蔡任,钱佳佳,许奇,许炜民,赵佩茹,袁嘉奇,李妍,黄桂成

  摘要:目的:通过观测游泳运动对膝骨关节炎(KOA)小鼠软骨基质降解及NF-质降信号通路介导的炎症反应的影响,探讨游泳运动延缓KOA小鼠软骨退变的分子机制。方法:3月龄C57BL/6小鼠40只,随机分为空白组(Blank)、ACLT模型组(ACLT)、ACLT模型+游泳组(ACLT+Swim)、假手术组(Sham)和假手术+游泳组(Sham+Swim)。采用前交叉韧带横断术(ACLT)制作KOA模型,手术2周后,游泳组小鼠进行6周的无负重游泳运动(40 min/d,5 d/周)。游泳结束后,取各组小鼠膝关节软骨,HE染色观察骨病理学改变,免疫组织化学染色检测软骨中炎症介质表达、RT-PCR法检测软骨基质相关指标mRNA表达、Western blot技术检测软骨基质及NF-κB通路相关分子蛋白表达,ELISA方法检测血清中炎症指标表达水平。结果:1)与空白组相比:ACLT组软骨损伤明显,Mankin’s评分显著升高(P<0.01);软骨Col II含量显著下降,而MMP-13表达增高(P<0.01);软骨中炎症介质COX-2、iNOS表达增高,血清炎症因子TNF-α、IL-6表达亦增高(P<0.01);NF-κB通路中磷酸化p65、IκBα蛋白水平显著升高(P<0.01)。2)与ACLT组相比:ACLT+Swim组Mankin′s评分明显下降(P<0.01)Col IImRNA(P<0.05)及蛋白水平(P<0.01)均提升,MMP-13mRNA及蛋白水平均显著下降(P<0.01),软骨中炎症介质COX-2、iNOS及血清炎症因子TNF-α、IL-6均显著降低(P<0.01),p65、IκBα磷酸化水平亦明显下降(P<0.01)。结论:中等强度游泳运动可能通过调控NF-κB信号通路介导的炎症反应,抑制软骨基质降解从而延缓软骨退变,干预KOA小鼠病理进程。
  关键词:游泳运动;膝骨关节炎;软骨退变;基质降解;炎症反应;核转录因子κB信号通路
  中图分类号:G804.53文献标识码:A文章编号:1006-2076(2021)06-0025-09
  Effects of swimming on cartilage degeneration and NF-κB activation in mice with knee osteoarthritis
  CAI Ren1,QIAN Jiajia2,3,XU Qi2,XU Weimin2, ZHAO Peiru3,YUAN Jiaqi3,LI Yan3, HUANG Guicheng2
  1.Dept.of Basic Physical Education,Nanjing University of Chinese Medicine,Nanjing 210023,Jiangsu,China;2.Laboratory for New Techniques of Restoration & Reconstruction of Orthopedics and Traumatology, Nanjing 210023, Jiangsu,China;3.Dept. of Rehabilitation Therapy, Nanjing University of Chinese Medicine, Nanjing 210023, Jiangsu,China
  Abstract:Objective:To observe the effect of swimming exercise with moderate intensity on cartilage matrix degradation and inflammatory response mediated by NF-κB signaling pathway in KOA mice, and to explore the molecular mechanism of swimming exercise delaying cartilage degeneration in KOA mice. Methods: 40 C57BL/6 mice aged 3 months were randomly divided into blank group (Blank), ACLT model group (ACLT), ACLT+swim group (ACLT+Swim), sham operation group (Sham) and sham+swim group (Sham+Swim). KOA model was made by anterior cruciate ligament transection (ACLT), 2 weeks after the operation, mice in the swim group were swimming without weight for 6 weeks (40 min/D, 5d/week). After swimming, the cartilage of knee joint of mice in each group was taken, and the pathological changes of cartilage were observed by HE staining. The expression of inflammatory mediators in cartilage was detected by immunohistochemical staining. The mRNA expression of cartilage matrix related indicators was detected by RT-PCR. The protein expression of cartilage matrix and NF-κB pathway related molecules was detected by Western blot. The expression level of inflammatory indicators in serum was detected by ELISA. Results: 1)Compared with the blank group, the cartilage injury in ACLT group was obvious, Mankin's score was significantly increased (P<0.01); the content of Col II in cartilage was significantly decreased, but the expression of MMP-13 was increased (P<0.01); the expression of COX-2 and iNOS in cartilage was increased, and the expression of TNF-α and IL-6 in serum was also increased (P<0.01); the protein levels of phosphorylated p65 and IκBα in NF-κB pathway were significantly increased (P<0.01). 2) Compared with ACLT group, Mankin's score decreased significantly (P<0.01), Col IImRNA (P<0.05) and protein level (P<0.01) were both increased, MMP-13 mRNA and protein level decreased significantly (P<0.01), COX-2, iNOS in cartilage and TNF-α, IL-6 in serum decreased significantly (P<0.01), the level of p65 and IκBα phosphorylation decreased significantly (P<0.01) in ACLT+Swim group. Conclusion: Moderate intensity swimming can inhibit the degradation of cartilage matrix by regulating the inflammatory response mediated by NF-κB signaling pathway, thus delaying cartilage degeneration and intervening the pathological process of KOA mice. This study provides a theoretical and experimental basis for swimming assisted therapy of KOA.

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