Laboratory of Periodontal Physiology, Dentistry Academic Unit, Faculty of Medicine, Pontificia Universidad Católica de Chile, Santiago, Chile.
J Periodontal Res. 2011 Aug;46(4):456-63. doi: 10.1111/j.1600-0765.2011.01360.x. Epub 2011 Mar 13.
Statins have been used to control hypercholesterolemia. However, these drugs also exert pleiotropic effects that include the modulation of inflammation and cell signaling. The present study has analyzed the effects of simvastatin on several cell responses involved in tissue repair, including cell adhesion, cell migration and invasion, actin cytoskeleton remodeling and cell viability.
Primary cultures of gingival fibroblasts were stimulated with simvastatin. Cell adhesion was evaluated using a colorimetric assay. Cell spreading was evaluated microscopically. Cell migration and invasion were assessed using a scratch wound-healing assay and a bicameral cell culture system, respectively. Changes in actin cytoskeleton and focal adhesion assembly were evaluated through immunofluorescence for actin, vinculin and active β1 integrin. Rac activation was evaluated by means of a pull-down assay. Cell viability was assessed using a colorimetric assay that determines mitochondrial functionality. Data analysis was performed using the Mann-Whitney U-test.
Simvastatin diminished cell adhesion and spreading over a fibronectin matrix. It also altered the closure of scratch wounds induced on cell monolayers and cell invasion through a Transwell system. Simvastatin-treated cells displayed an altered lamellipodia with poorly developed focal adhesion contacts and reduced levels of β1 integrin activation. During cell spreading, simvastatin diminished Rac activation.
The present study shows that simvastatin may alter cell migration by disrupting the cell signaling networks that regulate the actin cytoskeleton dynamics. This mechanism may affect the response of gingival mesenchymal cells during wound healing.
他汀类药物已被用于控制高胆固醇血症。然而，这些药物还具有多种作用，包括调节炎症和细胞信号转导。本研究分析了辛伐他汀对几种参与组织修复的细胞反应的影响，包括细胞黏附、细胞迁移和侵袭、肌动蛋白细胞骨架重塑和细胞活力。
用辛伐他汀刺激原代培养的牙龈成纤维细胞。通过比色法评估细胞黏附。通过显微镜评估细胞铺展。通过划痕愈合试验和双室细胞培养系统分别评估细胞迁移和侵袭。通过肌动蛋白、纽蛋白和活性β1 整合素的免疫荧光评估肌动蛋白细胞骨架和焦点黏附组装的变化。通过下拉测定评估 Rac 激活。通过测定线粒体功能的比色法评估细胞活力。使用曼-惠特尼 U 检验进行数据分析。
辛伐他汀降低了细胞在纤维连接蛋白基质上的黏附和铺展。它还改变了细胞单层上划痕伤口的闭合和 Transwell 系统中的细胞侵袭。用辛伐他汀处理的细胞显示出改变的片状伪足，焦点黏附接触不良，β1 整合素激活水平降低。在细胞铺展过程中，辛伐他汀降低了 Rac 的激活。
本研究表明，辛伐他汀可能通过破坏调节肌动蛋白细胞骨架动力学的细胞信号转导网络来改变细胞迁移。这种机制可能会影响牙龈间充质细胞在伤口愈合过程中的反应。
