Volk Susan W, Kapatkin Amy S, Haskins Mark E, Walton Raquel M, D'Angelo Marina
Department of Clinical Studies, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104-6044, USA.
Am J Vet Res. 2003 Oct;64(10):1225-33. doi: 10.2460/ajvr.2003.64.1225.
To determine matrix metalloproteinase (MMP) activity in synovial fluid (SF) obtained from the joints of dogs with degenerative joint disease (DJD) secondary to various underlying conditions.
35 samples of SF obtained from 18 clinically normal (control) dogs and 34 samples of SF obtained from 17 dogs with DJD; dogs with DJD were from 2 populations (client-owned dogs and research dogs that had DJD secondary to the lysosomal storage disease mucopolysaccharidosis VII).
MMP activity in samples of SF was semiquantitatively examined by use of gelatin or casein zymography. Western blot analysis was performed by use of antibodies for MMP-2 and MMP-9. In addition, in situ MMP activity was observed in sections of synovial membrane obtained from healthy and osteoarthritic joints.
Samples of SF from osteoarthritic joints had higher MMP-2 activity and dramatically increased MMP-9 activity, compared with values for healthy joints. Substrate-overlay analyses indicated minimal gelatin-degrading activity in synoviocytes obtained from control dogs, whereas greater activity was seen in osteoarthritic synoviocytes, with additional activity in the underlying tissue.
Higher MMP-2 activity and dramatic increases in MMP-9 activity were associated with the osteoarthritic state, even though MMP-2 activity was detected in healthy joints. This study expands information on MMP production in SF of osteoarthritic joints in other species and documents the similarity of MMP activity patterns regardless of the cause of DJD.
测定继发于各种潜在病症的退行性关节病(DJD)犬关节滑液(SF)中的基质金属蛋白酶(MMP)活性。
从18只临床正常(对照)犬获取35份SF样本,从17只患有DJD的犬获取34份SF样本;患有DJD的犬来自2个群体(客户拥有的犬以及因溶酶体贮积病粘多糖贮积症VII继发DJD的实验犬)。
通过明胶或酪蛋白酶谱法对SF样本中的MMP活性进行半定量检测。使用针对MMP-2和MMP-9的抗体进行蛋白质印迹分析。此外,在从健康和骨关节炎关节获取的滑膜切片中观察原位MMP活性。
与健康关节相比,骨关节炎关节的SF样本具有更高的MMP-2活性以及显著增加的MMP-9活性。底物覆盖分析表明,从对照犬获取的滑膜细胞中明胶降解活性最低,而在骨关节炎滑膜细胞中观察到更高的活性,并且在其下方组织中也有额外活性。
尽管在健康关节中检测到MMP-2活性,但更高的MMP-2活性以及MMP-9活性的显著增加与骨关节炎状态相关。本研究扩展了关于其他物种骨关节炎关节SF中MMP产生的信息,并证明了无论DJD病因如何,MMP活性模式的相似性。