Reduction in lipopolysaccharide-induced apoptosis of fibroblasts obtained from a patient with gingival overgrowth during nifedipine-treatment
Reduction in lipopolysaccharide-induced apoptosis of fibroblasts obtained from a patient with gingival overgrowth during nifedipine-treatment
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DOI：
10.1016/j.archoralbio.2011.03.006
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发表时间：
2011-10-01
影响因子：
3
通讯作者：
Fujii, Akira
中科院分区：
文献类型：
--
作者：
Takeuchi, Reiri;Matsumoto, Hiroko;Fujii, Akira
目的：我们之前已经证明硝苯地平（NIF）诱导牙龈过度生长的机制与观察到NIF反应性患者（NIFr）的牙龈成纤维细胞增殖和细胞周期进展大于NIF非反应性患者（NIFn）有关。据报道，牙龈过度生长也是牙龈成纤维细胞凋亡受到抑制的结果。脂多糖（LPS）可诱导成纤维细胞凋亡。因此，我们重点评估NIFn和NIFr在lps诱导的细胞凋亡方面是否存在差异。方法：将NIFn和NIFr捕获在含有0.5% FBS的DMEM中，用LPS刺激，检测细胞凋亡、细胞周期分析、Western blotting和caspase活性。结果：与NIFn相比，NIFr的凋亡细胞数量明显减少，S期和G(2)/M期细胞比例明显增加。与NIFn相比，LPS没有上调NIFr中Bax和细胞色素c蛋白的水平。NIFn和NIFr的蛋白表达变化如下：Bad升高，Bcl-xL降低，Bcl-2和p53不变。LPS显著提高了NIFn中Caspase-3和-9的活性，但在NIFr中没有变化。在LPS处理下，NIFn和NIFr的Caspase-2活性保持不变，而caspase-8活性显著增加。结论：Bad、Bax、细胞色素c、p53、caspase -2、-3、-8、-9是促凋亡蛋白。Bcl-2和Bcl-xL是抗凋亡蛋白。因此，nif诱导牙龈过度生长的机制可能与NIFr中Bax、细胞色素c、caspase-3和-9的减少有关。(C) 2011 Elsevier Ltd.版权所有。
Objective: We have previously demonstrated that the mechanism of nifedipine (NIF)-induced gingival overgrowth is related to the observation that proliferation and cell cycle progression of gingival fibroblasts derived from NIF reactive patient (NIFr) are greater than those from NIF non-reactive patient (NIFn). Gingival overgrowth has also been reported to be a result of inhibited apoptosis of gingival fibroblasts. Apoptosis in fibroblasts is induced by lipopolysaccharide (LPS). Thus, we focused upon evaluating whether there is a difference in LPS-induced apoptosis between NIFn and NIFr.Methods: Both NIFn and NIFr were arrested in DMEM containing 0.5% FBS, stimulated by LPS, and assayed for apoptosis, cell cycle analysis, Western blotting, and caspase activity.Results: Compared to NIFn, the number of apoptotic cells was significantly decreased and the percentage of cells in S and G(2)/M phase was significantly increased in NIFr. The levels of Bax and cytochrome c proteins in NIFr were not up-regulated by LPS compared with NIFn. Both NIFn and NIFr displayed the following changes in protein expression: increased Bad, decreased Bcl-xL, and unchanged Bcl-2 and p53. Caspase-3 and -9 activities were significantly increased by LPS in NIFn but were unchanged in NIFr. Caspase-2 activity remained constant whilst caspase-8 activity significantly increased upon LPS treatment in both NIFn and NIFr.Conclusion: Bad, Bax, cytochrome c, p53, and caspases-2, -3, -8, and -9 are pro-apoptotic proteins. Bcl-2 and Bcl-xL are anti-apoptotic proteins. Thus, the mechanism of NIF-induced gingival overgrowth might be related to decreased apoptosis in NIFr through a reduction of Bax, cytochrome c, and caspase-3 and -9. (C) 2011 Elsevier Ltd. All rights reserved.