Oral malodorous compound triggers mitochondrial-dependent apoptosis and causes genomic DNA damage in human gingival epithelial cells
Oral malodorous compound triggers mitochondrial-dependent apoptosis and causes genomic DNA damage in human gingival epithelial cells
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DOI：
10.1111/j.1600-0765.2008.01199.x
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发表时间：
2010-02-01
影响因子：
3.5
通讯作者：
Ii, H.
中科院分区：
文献类型：
--
作者：
Calenic, B.;Yaegaki, K.;Ii, H.
背景和目的：挥发性硫化合物是引起口臭的主要化合物。其中一种化合物硫化氢 (H2S) 是造成生理性口臭的原因，据报道也具有牙周致病活性。硫化氢已被证明可以激活不同组织中的细胞凋亡过程。细胞凋亡在牙周炎的发生发展中起着重要作用。本研究的目的是确定 H2S 是否会导致人牙龈上皮细胞凋亡，并检查启动该过程的细胞信号通路。 材料和方法：将人牙龈上皮细胞与 50 ng/mL H2S 在含有 5% CO2 的空气中孵育 24、48 或 72 小时。为了检测细胞凋亡，用膜联蛋白 V 和 7-氨基放线菌素 D 对细胞进行染色，并使用流式细胞术进行分析。使用流式细胞术和酶联免疫吸附测定评估活性氧、线粒体膜去极化和细胞色素 C 释放到胞质溶胶中。还测定了关键凋亡酶 caspase-9、-8 和 -3 的活性水平。使用单细胞凝胶电泳检测基因组 DNA 损伤。结果：24 小时时细胞凋亡显着增加至 24.5 +/- 5.7，48 小时时显着增加至 41.5 +/- 8.9%（p < 0.01）。活性氧增强，线粒体膜去极化崩溃。细胞色素 C 释放显着增加（24 小时时为 0.12 +/- 0.02 与 0.02 +/- 0.01，48 小时时为 0.21 +/- 0.02 与 0.02 +/- 0.01 ng/mL；p < 0.05）。 Caspase-9 和-3 被强烈激活，而 caspase-8 活性仍然较低。 DNA链断裂百分比增加，特别是在48小时时。结论：硫化氢通过激活线粒体途径诱导人牙龈上皮细胞凋亡。
Background and Objective: Volatile sulfur compounds are the main compounds causing halitosis. One of these compounds, hydrogen sulfide (H2S), which is responsible for physiological halitosis, is reported also to have periodontal pathogenic activities. Hydrogen sulfide has been shown to activate the apoptotic process in different tissues. Apoptosis plays an important role in the development of periodontitis. The aim of this study was to determine whether H2S causes apoptosis in human gingival epithelial cells and to examine the cellular signaling pathway initiating the process.Material and Methods: Human gingival epithelial cells were incubated with 50 ng/mL H2S in air contining 5% CO2 for 24, 48 or 72 h. To detect apoptosis, the cells were stained with annexin V and 7-amino actinomycin D, and analyzed using flow cytometry. Reactive oxygen species, mitochondrial membrane depolarization and release of cytochrome C into the cytosol were assessed using flow cytometry and enzyme-linked immunosorbent assay. Activity levels for the key apoptotic enzymes caspase-9, -8 and -3 were also determined. Genomic DNA damage was detected using single-cell gel electrophoresis.Results: Apoptosis was significantly increased to 24.5 +/- 5.7 at 24 h and 41.5 +/- 8.9% at 48 h (p < 0.01). Reactive oxygen species were enhanced and mitochondrial membrane depolarization was collapsed. Cytochrome C release was dramatically increased (0.12 +/- 0.02 vs. 0.02 +/- 0.01 at 24 h and 0.21 +/- 0.02 vs. 0.02 +/- 0.01 ng/mL at 48 h; p < 0.05). Caspase-9 and -3 were strongly activated, while caspase-8 activity remained low. The percentage of DNA strand breaks increased, especially at 48 h.Conclusion: Hydrogen sulfide induces apoptosis in human gingival epithelial cells by activating the mitochondrial pathway.