Jackson C J, Arkell J, Nguyen M
Sutton Arthritis Research Laboratory, Department of Rheumatology, Royal North Shore Hospital, St Leonards NSW, Australia.
Ann Rheum Dis. 1998 Mar;57(3):158-61. doi: 10.1136/ard.57.3.158.
Angiogenesis (the formation of new blood vessels) is a major component of the inflammatory pannus in rheumatoid arthritis (RA). Matrix metalloproteinase (MMP) secretion by microvascular endothelial cells is an essential step in angiogenesis. The secretion of MMP1, MMP2, MMP9, and TIMP1 by human microvascular endothelial cells derived from RA synovium (RASE) to normal synovium (NSE) and neonatal foreskin (FSE) was compared.
Confluent monolayers of endothelial cells in basal medium were pre-incubated for 24 hours in the presence or absence of phorbol myristate acetate (PMA, 100 ng/ml). MMP1 activity was measured using a spectrophotometric assay and western blotting. MMP2 and MMP9 were measured using zymography. TIMP1 was measured by enzyme linked immunosorbent assay and western blotting.
There was little difference between the amounts of MMP2 secreted by any of the cell lines. In response to PMA both synovial cell types showed a significantly higher MMP1 and MMP9 activity compared with FSE, although there was no difference between RASE and NSE. Tumour necrosis factor alpha had minimal effect on MMP activity. There was a striking decrease in the amount of TIMP1 secreted by RASE compared with normal synovium.
As overall MMP activity is a balance between the amount of MMP and TIMP1 present, the low levels of TIMP1 produced by RASE would shift the balance in favour of increased MMP activity by these cells. This is likely to contribute to the angiogenic potential of RASE.
血管生成（新血管的形成）是类风湿关节炎（RA）炎症性血管翳的主要组成部分。微血管内皮细胞分泌基质金属蛋白酶（MMP）是血管生成的关键步骤。比较了类风湿关节炎滑膜来源的人微血管内皮细胞（RASE）、正常滑膜来源的人微血管内皮细胞（NSE）和新生儿包皮来源的人微血管内皮细胞（FSE）分泌MMP1、MMP2、MMP9和TIMP1的情况。
将处于基础培养基中的汇合内皮细胞单层在有或无佛波酯（PMA，100 ng/ml）存在的情况下预孵育24小时。使用分光光度法和蛋白质印迹法测量MMP1活性。使用酶谱法测量MMP2和MMP9。通过酶联免疫吸附测定法和蛋白质印迹法测量TIMP1。
任何细胞系分泌的MMP2量之间几乎没有差异。与FSE相比，在PMA刺激下，两种滑膜细胞类型的MMP1和MMP9活性均显著更高，尽管RASE和NSE之间没有差异。肿瘤坏死因子α对MMP活性影响最小。与正常滑膜相比，RASE分泌的TIMP1量显著减少。
由于总体MMP活性是MMP和TIMP1量之间的平衡，RASE产生的低水平TIMP1会使平衡向有利于这些细胞增加MMP活性的方向转变。这可能有助于RASE的血管生成潜力。
