摘要
目的观察中医手法对实验性兔膝骨关节炎软骨退变的干预作用,并从细胞信号转导角度探讨其作用机制。方法 24只新西兰大白兔随机分为3组,即正常组、模型组和手法组,除正常组外,其余两组动物均采用手术方法建立膝骨关节炎模型。造模4周后对手法组给予髌骨按揉、关节被动屈伸等形式的手法治疗。干预8周后处死动物,暴露膝关节行形态学观察,并取股骨髁部做病理切片,行HE染色,镜下观察其组织病理学特征并进行Mankin’s评分,比较各组关节软骨退变情况。应用RT-PCR方法分别检测滑膜组织中IL-1β、i NOS、MMP-13和软骨组织中P38、ERK1/2的mRNA表达水平。应用Western blot方法检测软骨组织内MMP-13及磷酸化的ERK1/2(p-ERK1/2)的蛋白表达。比较各组实验数据并进行统计学分析。结果手法组关节软骨退变程度较模型组明显减轻,Mankin’s评分分值显著低于模型组(P<0.05)。手法组软骨P38、ERK1/2及滑膜组织IL-1β、i NOS、MMP-13的mRNA表达水平较模型组显著下调(P<0.05)。模型组和手法组的MMP-13、p-ERK1/2的蛋白表达较正常关节软骨明显增强,而手法组MMP-13、p-ERK1/2的蛋白活化程度与模型组比较明显受到抑制。结论中医手法能够有效抑制膝骨关节炎的软骨退变程度,影响炎症介质和细胞因子的表达,其机制可能是通过抑制软骨MAPK信号转导通路发挥作用。
Objective To observe the intervention effects of Chinese medical manipulation on the articular cartilage degeneration in rabbit model of knee osteoarthritis,and explore its mechanism from the perspective of cell signaling transduction.Methods 24 New Zealand white rabbits were randomly divided into the normal group,model group and manipulation group.Except the normal group,the model of knee osteoarthritis was established by operation in the other two groups.After modeling for 4 weeks,the manipulation group was treated with manipulation therapy such as patella massage and joint passive flexion and extension.After intervention for 8 weeks,the rabbits were sacrificed and the knee joints were exposed for morphological observation.The femoral condyle was collected and made into pathological sections for HE staining.The histopathological features were observed under microscope and evaluated by Mankin ＇s score,and the degeneration of articular cartilage in each group was compared.The mRNA expression levels of IL-1β、i NOS and MMP-13 in synovial tissues and P38 as well as ERK1/2 in cartilage tissue were detected by RT-PCR.The protein expression levels of MMP-13 and phosphorylated ERK1/2（p-ERK1/2） in cartilage tissue were detected by Western-blot.All the experimental data were compared and conducted the statistical analysis.Results The degree of articular cartilage degeneration in the manipulation group was obviously alleviated compared with the model group,and the Mankin＇s score in the manipulation group was significantly lower than that in the model group（P〈0.05）.Compared with the model group,the mRNA expression levels of P38 and ERK1/2 in cartilage and IL-1β,i NOS and MMP-13 in synovial tissues in the manipulation group were significantly down-regulated（P〈0.05）.Compared with the normal articular cartilage,the protein expressions of MMP-13 and p-ERK1/2 were significantly up-regulated in the model group and manipulation group.While the protein activation degree of MMP-13 and p-ERK1/2 in the manipula
作者
杨松滨
刘益杰
冯伟
吴绪波
黄正
胡笑燊
褚立希
YANG Songbin;LIU Yijie;FENG Wei;WU Xubo;HUANG Zheng;HU Xiaoshen;CHU Lixi(School of Rehabilitation Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China;Department of Spinal Surgery, Guanghua Hospital of Integrated Chinese and Western Medicine, Changning District, Shanghai 200052, China)
出处
《上海中医药杂志》
2018年第6期72-77,共6页
Shanghai Journal of Traditional Chinese Medicine
基金
国家自然科学基金资助项目(81173357)
上海中医药大学研究生创新培养项目(Y201823)
上海中医药大学康复医学院岐康青年创新人才计划项目(A4-Q1501807)
关键词
膝骨关节炎
手法
关节软骨
丝裂原活化蛋白激酶
信号通路
knee osteoarthritis
manipulation
articular cartilage
mitogen- activated protein kinase
signaling pathway
