Carvalho Rodrigo V, Ogliari Fabrício A, de Souza Ana P, Silva Adriana F, Petzhold Cesar L, Line Sergio R P, Piva Evandro, Etges Adriana
Department of Operative Dentistry, School of Dentistry, Federal University of Pelotas, Rio Grande do Sul, Brazil.
Eur J Oral Sci. 2009 Feb;117(1):64-7. doi: 10.1111/j.1600-0722.2008.00591.x.
This study evaluated the effect of different concentrations of 2-hydroxyethyl methacrylate (HEMA) on the inhibition of matrix metalloproteinase-2 (MMP-2) in vitro. Mouse gingival explants were cultured overnight in Dulbecco's modified Eagle's minimal essential medium, following which the expression of secreted enzymes was analyzed by gelatin zymography and the effects of different amounts of HEMA on enzyme activity were investigated. The gelatinolytic proteinases present in the conditioned media were characterized as being matrix metalloproteinases (MMPs) by means of specific chemical inhibition. The MMPs present in the conditioned media were identified, using immunoprecipitation, as MMP-2. Three major bands were detected in the zymographic assays and were characterized, according to their respective molecular weights, into the following forms of MMP-2: zymogene (72 kDa), intermediate (66 kDa), and active (62 kDa). All forms of MMP-2 were inhibited by HEMA in a dose-dependent manner, implying that MMP-2 may be inhibited by HEMA in vivo.
本研究评估了不同浓度的甲基丙烯酸羟乙酯（HEMA）在体外对基质金属蛋白酶-2（MMP-2）的抑制作用。将小鼠牙龈外植体在杜氏改良 Eagle 最低必需培养基中培养过夜，随后通过明胶酶谱法分析分泌酶的表达，并研究不同量的 HEMA 对酶活性的影响。通过特异性化学抑制法，将条件培养基中存在的明胶水解蛋白酶鉴定为基质金属蛋白酶（MMPs）。使用免疫沉淀法将条件培养基中存在的 MMPs 鉴定为 MMP-2。在酶谱分析中检测到三条主要条带，并根据其各自的分子量将其表征为以下形式的 MMP-2：酶原（72 kDa）、中间产物（66 kDa）和活性形式（62 kDa）。所有形式的 MMP-2 均被 HEMA 以剂量依赖性方式抑制，这意味着 MMP-2 在体内可能被 HEMA 抑制。
