Expression of matrix metalloproteinases and inhibitor by human trabecular meshwork.
Expression of matrix metalloproteinases and inhibitor by human trabecular meshwork.
复制标题
DOI：
--
复制
发表时间：
1991
影响因子：
4.4
通讯作者：
J. Alexander;J. Samples;E. Michael;Van Duskirk;Ted Acotr
中科院分区：
文献类型：
--
作者：
J. Alexander;J. Samples;E. Michael;Van Duskirk;Ted Acotr
细胞外基质（ECM）的转换和重塑至少部分是由基质金属蛋白酶家族成员的调节分泌启动的。人和牛小梁网状组织在培养中分泌间质胶原酶，包括72-和92-kD形式的IV型胶原酶（明胶酶）和基质溶解素，以及金属蛋白酶的组织抑制剂（TIMP）。这些蛋白酶和TIMP通过免疫印迹法从十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）中转移，使用几种特异性的抗蛋白和抗肽多克隆抗体进行鉴定。明胶酶和基质溶酶的酶活性也通过底物SDS-PAGE进行了分析，其中蛋白酶底物在电泳之前被聚合到凝胶中，以便进行后续的活性测定。这些基质金属蛋白酶和TIMP在小梁培养基中以低基础水平分泌；用phorbol有丝分裂原处理培养物，它们的分泌水平增加了几倍。12-O-tetradecanoylphorbol-13-acetate (TPA)。小梁基质金属蛋白酶和TIMP的特征与包括视网膜色素上皮在内的许多其他组织分泌的金属蛋白酶和TIMP相似。这些蛋白酶可能在正常和青光眼的小梁细胞外基质以及随后的房水流出通道的维持和调节中起重要作用。
Extracellular matrix (ECM) turnover and remodeling are initiated, at least in part, by the regulated secretion of members of a family of matrix metalloproteinases. Human and bovine trabecular mesh-work in culture secrete interstitial collagenase, both the 72- and the 92-kD forms of type IV collagenase (gelatinases) and stromelysin, and the tissue inhibitor of metalloproteinases (TIMP). These proteinases and TIMP were identified by immunoblotting western transfers from sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using several specific antiprotein and antipeptide polyclonal antibodies. Gelatinase and stromelysin enzymatic activities were also analyzed by substrate SDS-PAGE, in which proteinase substrates were polymerized into the gels before electrophoresis to allow subsequent activity assays. These matrix metalloproteinases and TIMP are secreted at low basal levels into trabecular culture medium; their secretion levels are increased several-fold by treatment of the cultures with the phorbol mitogen. 12-O-tetradecanoylphorbol-13-acetate (TPA). Characteristics of the trabecular matrix metalloproteinases and TIMP are similar to those secreted by numerous other tissues, including the retinal pigment epithelium. These proteinases may serve an important role in the maintenance and regulation of the trabecular extracellular matrix and, subsequently, of the aqueous humor outflow pathway in normal and glaucomatous eyes.