Department of Oral Biology, Indiana University School of Dentistry, Indianapolis, IN 46202, USA.
J Periodontol. 2011 Jul;82(7):1071-9. doi: 10.1902/jop.2010.100540. Epub 2010 Dec 13.
Members of the matrix metalloproteinase (MMP) family have been shown to be involved in periodontal disease. Risk factors for periodontal disease include tobacco smoking. Cigarette smoke condensate (CSC) is comprised of thousands of chemicals. Nicotine is one of the active components in tobacco. This study compares the effects of CSC and nicotine at the level in CSC on the collagen-degrading ability of human gingival fibroblasts (HGFs) and the expression of selected MMPs and tissue inhibitors of metalloproteinases (TIMPs).
HGFs were seeded in six-well collagen-coated plates, exposed to 100 μg/mL (2.4 μg/mL nicotine) of CSC or 2.4 μg/mL nicotine for 3 days, and then collagen degradation was analyzed. After 3 days exposure to CSC or nicotine, the conditioned media from HGFs was collected and the membrane proteins were extracted for gelatin zymography and Western blot analyses. The mRNA levels of MMP-2, MMP-14, and TIMP-2 were measured by reverse transcription-polymerase chain reaction.
The CSC increased collagen degradation, and increased the levels of TIMP-2, MMP-14, and the active MMP-2 in the membrane extracts, and their mRNA levels. CSC also increased the level of active MMP-2 in the conditioned media. Nicotine at the level in CSC (2.4 μg/mL) had little influence on collagen degradation, as well as on the protein and mRNA levels of MMP-2, MMP-14, and TIMP-2.
CSC may increase HGF-mediated collagen degradation by affecting membrane-associated MMPs and TIMPs.
基质金属蛋白酶（MMP）家族的成员已被证明与牙周病有关。牙周病的危险因素包括吸烟。香烟烟雾冷凝物（CSC）由数千种化学物质组成。尼古丁是烟草中的一种活性成分。本研究比较了 CSC 中与 CSC 水平相当的尼古丁对人牙龈成纤维细胞（HGFs）胶原降解能力和选定的 MMPs 和金属蛋白酶组织抑制剂（TIMPs）表达的影响。
将 HGFs 接种在六孔胶原包被的培养板中，用 100μg/ml（2.4μg/ml 尼古丁）的 CSC 或 2.4μg/ml 尼古丁处理 3 天，然后分析胶原降解情况。用 CSC 或尼古丁处理 3 天后，收集 HGFs 的条件培养基，并提取膜蛋白进行明胶酶谱和 Western blot 分析。通过逆转录-聚合酶链反应测量 MMP-2、MMP-14 和 TIMP-2 的 mRNA 水平。
CSC 增加了胶原降解，并增加了膜提取物中 TIMP-2、MMP-14 和活性 MMP-2 的水平及其 mRNA 水平。CSC 还增加了条件培养基中活性 MMP-2 的水平。CSC 水平（2.4μg/ml）的尼古丁对胶原降解以及 MMP-2、MMP-14 和 TIMP-2 的蛋白和 mRNA 水平影响不大。
CSC 可能通过影响膜相关的 MMPs 和 TIMPs 增加 HGF 介导的胶原降解。
