Johst Ursula, Betsch Angelika, Wiskirchen Jakub, Schöber Wolfgang, Vonthein Reinhard, Rinkert Natalie, Kehlbach Rainer, Claussen Claus D, Duda Stephan H
Department of Diagnostic Radiology, Eberhard-Karls University, Tübingen, Germany.
J Cardiovasc Pharmacol. 2003 Apr;41(4):526-35. doi: 10.1097/00005344-200304000-00004.
The aim of this study was to evaluate the effects of 9-cis retinoid acid (9-cis RA) and all-trans RA (ATRA) on proliferation, migratory ability, synthesis of extracellular matrix, intracellular signal transduction, and differentiation of human aortic smooth muscle cells (haSMCs) in vitro. Changes of cell proliferation following incubation with RAs in different doses (10-6 M, 10-7 M, and 10-8 M) were determined directly by proliferation kinetics and indirectly by bromodeoxyuridine enzyme-linked immuno sorbant assays and colony-formation assays. The migratory ability of haSMCs was examined with the help of migration assays. The production of the extracellular matrix protein tenascin was explored by immunostaining. The amounts of total p44/p42 mitogen-activated protein kinases (MAPKs) and their phosphorylated forms were detected with the help of Western blots. To judge the state of differentiation of haSMCs, cell cycle distribution and the pattern of alpha-actin were analyzed. Both RAs clearly inhibited the proliferation of haSMCs in a dose-dependent manner. 9-cis RA had a tendency to be more effective than ATRA. After treatment with RAs, the migratory ability was especially reduced during stimulation with platelet-derived growth factor (PDGF) and the synthesis of tenascin decreased. Although the total p44/p42 MAPKs were downregulated, the amounts of activated forms increased markedly in the cells incubated with RAs and particularly stimulated with PDGF. The cell-cycle analysis demonstrated an increased G1-phase, complemented by a stronger expression of alpha-actin after treatment. 9-cis RA especially has the potential to inhibit the proliferation, migration, and synthesis of extracellular matrix of haSMCs by inducing differentiation in vitro.
本研究的目的是评估9-顺式视黄酸（9-cis RA）和全反式视黄酸（ATRA）对人主动脉平滑肌细胞（haSMCs）体外增殖、迁移能力、细胞外基质合成、细胞内信号转导及分化的影响。通过增殖动力学直接测定不同剂量（10-6 M、10-7 M和10-8 M）视黄酸孵育后细胞增殖的变化，并通过溴脱氧尿苷酶联免疫吸附测定和集落形成测定间接测定。借助迁移试验检测haSMCs的迁移能力。通过免疫染色探索细胞外基质蛋白腱生蛋白的产生。借助蛋白质印迹法检测总p44/p42丝裂原活化蛋白激酶（MAPKs）及其磷酸化形式的含量。为判断haSMCs的分化状态，分析细胞周期分布和α-肌动蛋白模式。两种视黄酸均明显以剂量依赖性方式抑制haSMCs的增殖。9-顺式视黄酸比全反式视黄酸更具效果倾向。用视黄酸处理后，在血小板衍生生长因子（PDGF）刺激期间迁移能力尤其降低，腱生蛋白合成减少。尽管总p44/p42 MAPKs下调，但在用视黄酸孵育并特别用PDGF刺激的细胞中，活化形式的含量显著增加。细胞周期分析显示G1期增加，处理后α-肌动蛋白表达增强。9-顺式视黄酸尤其具有通过体外诱导分化来抑制haSMCs增殖、迁移和细胞外基质合成的潜力。
