摘要
目的:观察持续被动运动(CPM)和补充一氧化氮(NO)供体——硝酸甘油(NG)对兔骨关节炎(OA)模型中软骨基质金属蛋白酶-1(MMP-1)和MMP-13表达以及NO含量的影响,探讨CPM下调MMPs的可能机制.方法:30只3~4月龄雄性新西兰大白兔,其中6只进行假手术作为正常对照组(NC组),另外24只建立膝关节OA模型,术后随机分为4组,即OA对照组(OA组)、OA硝酸甘油给药组(NG组)、OA持续被动运动组(CPM组)、OA持续被动运动+硝酸甘油给药组(CPM+ NG组),每组各6只.NC组和OA组不作处理,NG组给予NG软膏膝关节局部涂抹,CMP组在CPM训练仪上进行膝关节持续被动运动,CMP+ NG组即在运动干预的同时给予NG局部涂抹.4周后取各组软骨组织,硝酸还原酶法测定NO含量,HE染色观察软骨形态学变化并进行Mankin's评分,实时荧光定量PCR(RQ-PCR)检测MMP-1和MMP-13 mRNA水平,免疫组织化学法测定MMP-1和MMP-13蛋白表达量.结果:NO含量、Mankin's评分以及MMP-1和MMP-13 mRNA和蛋白水平在OA组明显高于NC组(P<0.01),NG组高于OA组(P<0.01),CPM组低于OA组和NG组(P<0.01),CPM+ NG组低于NG组(P<0.01),但高于CPM组(P<0.01).结论:CPM通过抑制软骨细胞NO合成下调MMP-1和MMP-13表达,进而对软骨细胞起保护作用.
Objective: To observe the effects of continuous passive motion (CPM) and nitric oxide (NO) donor- nitroglycerin (NG) supplement on matrix metalloproteinase- 1 (MMP- 1 ), matrix metalloproteinase- 13 (MMP- 13 ) expression and NO content in cartilage of rabbit osteoarthritis (OA) model and investigate the possible mechanism that CPM downregulated MMPs. Methods : Of thirty male New Zealand white rabbits of 3 - 4 months, six sham operated served as normal control group (NC, not treated) and the other twenty- four animals being created OA model were randomly allocated four group: osteoar- thritis control group (OA, not treated), nitroglycerin group (NG, treated with NG ointment on knee joint), continuous pas- sive motion group ( CPM, exercise on CPM training apparatus) and continuous passive motion + nitroglycerin group ( CPM + NG, exercise plus NG supplement). After four weeks, NO content were measured by nitrate reductase method, cartilage mor- phologic variations by HE stain and Mankin's score, MMP - 1 and MMP - 13 mRNA by real -time fluorescent quantitation PCR (RQ -PCR) and the protein level by immunohistochemistry. Results: NO content, Mankin~ score, MMP- 1 and MMP - 13 mRNA and protein level in OA group were higher than those in NC group ( P 〈0.01 ) and in NG group higher than OA group, while the indexes above in CPM group were lower than OA and NG group and in CPM + NG group lower than NG group but higher than CPM group. Conclusion: CPM downregulated expression of MMP - 1 and MMP - 13 via inhibition of NO synthesis in chondroeytes, thus protecting chondroeytes in OA model.
出处
《广州体育学院学报》
北大核心
2015年第3期90-95,共6页
Journal of Guangzhou Sport University
关键词
持续被动运动
骨关节炎
软骨
一氧化氮
基质金属蛋白酶
continuous passive motion
osteoarthritis
cartilage
nitric oxide
matrix metalloproteinase