Oral malodorous compound causes apoptosis and genomic DNA damage in human gingival fibroblasts
Oral malodorous compound causes apoptosis and genomic DNA damage in human gingival fibroblasts
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DOI：
10.1111/j.1600-0765.2007.01052.x
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发表时间：
2008-08-01
影响因子：
3.5
通讯作者：
Kamoda, T.
中科院分区：
文献类型：
--
作者：
Yaegaki, K.;Qian, W.;Kamoda, T.
背景与目的：挥发性硫化物是引起口臭的主要原因。硫化氢是一种挥发性硫化物，是生理性口臭的主要恶臭化合物。此前已报道过硫化氢的周期性致病活性。硫化氢可诱导主动脉平滑肌细胞和其他组织的细胞凋亡。细胞凋亡在牙周炎的发生发展中起重要作用。本研究的目的是确定硫化氢是否会导致人牙龈成纤维细胞的凋亡。材料和方法：用乳酸脱氢酶法检测坏死细胞。用组蛋白复合DNA片段分析和流式细胞仪检测细胞凋亡。同时检测细胞凋亡信号转导关键酶caspase3的水平，以及硫化氢对细胞内活性氧和超氧化物歧化酶的影响。结果：100 ng/mLH_2S作用72 h后，只有不到10%的人牙龈成纤维细胞出现坏死，而细胞凋亡率显著增加(p<；0.05)。孵育48h和72h后，超氧化物歧化酶活性受到强烈抑制，活性氧产生增加。Caspase3活性在孵育72h后显著升高(p<；0.01)。单细胞凝胶电泳法测定的尾长、尾部脱氧核糖核酸百分率和尾矩在孵育72h后也明显增加(p<；0.001)。结论：硫化氢可引起人牙龈成纤维细胞的凋亡和DNA损伤。硫化氢刺激的活性氧水平升高可能诱导细胞凋亡和DNA链断裂。
Background and Objective: Volatile sulfur compounds are the main cause of halitosis. Hydrogen sulfide is one of these volatile sulfur compounds and the principal malodorous compound in physiological halitosis. Periodontally pathogenic activities of hydrogen sulfide have been previously reported. Hydrogen sulfide induces apoptotic cell death in aorta smooth muscle cells and in other tissues. Apoptosis plays an important role in the onset and progress of periodontitis. The objective of this study was to determine whether hydrogen sulfide causes apoptosis in human gingival fibroblasts.Material and methods: Necrotic cells were detected using a lactate dehydrogenase assay. Apoptosis was ascertained using a histone-complexed DNA fragment assay and flow cytometry. The level of caspase 3, a key enzyme in apoptotic signaling, was also measured, and the effects of hydrogen sulfide on reactive oxygen species and superoxide dismutase were assessed. DNA damage caused by hydrogen sulfide was examined by means of single-cell gel electrophoresis.Results: After 72 h of incubation with 100 ng/mL of hydrogen sulfide, necrosis was found in less than 10% of human gingival fibroblasts, whereas apoptosis was significantly increased (p < 0.05). Superoxide dismutase activity was strongly inhibited, and reactive oxygen species production was enhanced, after 48 and 72 h of incubation. Caspase 3 activity was also increased after 72 h of incubation (p < 0.01). Tail length, percentage of DNA in tail, and tail moment, measured by single-cell gel electrophoresis, were also intensified after 72 h of incubation (p < 0.001).Conclusion: Hydrogen sulfide caused apoptosis and DNA damage in human gingival fibroblasts. An increased level of reactive oxygen species stimulated by hydrogen sulfide may induce apoptosis and DNA strand breaks.