Flavin-containing enzymes as a source of reactive oxygen species in HEMA-induced apoptosis.
Flavin-containing enzymes as a source of reactive oxygen species in HEMA-induced apoptosis.
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含黄素酶作为 HEMA 诱导细胞凋亡中活性氧的来源
DOI：
10.1016/j.dental.2017.01.014
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Buchalla W
中科院分区：
文献类型：
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作者：
Schweikl H;Gallorini M;Forstner M;Petzel C;Bolay C;Hiller KA;Cataldi A;Krifka S;Buchalla W
由牙科复合材料的化合物如甲基丙烯酸2-羟乙酯（HEMA）诱导的氧化应激由于活性氧（ROS）的过量形成而干扰重要的细胞功能，导致细胞凋亡。暴露于树脂单体的细胞中ROS的来源是未知的。本研究探讨了含黄素的ROS和RNS的功能方法用小鼠RAW264.7巨噬细胞暴露于HEMA后，在用2′7′-二氯二氢荧光素二乙酸酯染色细胞后，通过流式细胞术（FACS）测定（0-6-8 mM（H2 DCF-DA）、二氢乙锭（DHE）或二氢罗丹明123（DHR 123）。通过Annexin-V-FITC/碘化丙啶标记，随后通过FACS分析鉴定凋亡或坏死的细胞。ROS/RNS产生酶的表达进行了分析，通过Western blotting.ResultsDCF荧光增加，在细胞暴露于HEMA为1小时，表明羟基自由基，H2 O2，或一氧化氮和超氧阴离子形成过氧亚硝酸根（ONOO-）的生产。24小时后增加的DHR 123荧光表明主要形成H2 O2。低浓度的二亚苯基碘鎓（DPI），含黄素酶的抑制剂，在HEMA的存在下，细胞凋亡的诱导减少。p47 phox的表达，超氧化物产生的Nox 2的调节亚基，下调，和NOS的表达，产生一氧化氮（NO）的可能是抑制的反馈环机制在HEMA暴露的文化。VAS 2870或夹竹桃黄素对HEMA诱导的细胞凋亡的抑制作用进一步表明了Nox 2的重要功能。
ObjectiveOxidative stress induced by compounds of dental composites like 2-hydroxyethyl methacrylate (HEMA) due to excess formation of reactive oxygen species (ROS) disturbs vital cell functions leading to apoptosis. The sources of ROS in cells exposed to resin monomers are unknown. The present study investigates functions of flavin-containing ROS and RNS (reactive nitrogen species) producing enzymes in cells exposed to HEMA.MethodsThe formation of oxidative stress in RAW264.7 mouse macrophages exposed to HEMA (0–6–8 mM) was determined by flow cytometry (FACS) after staining of cells with 2′7′-dichlorodihydrofluorescin diacetate (H2DCF-DA), dihydroethidium (DHE) or dihydrorhodamine 123 (DHR123). Cells in apoptosis or necrosis were identified by annexin-V-FITC/propidium iodide labeling followed by FACS analysis. Expression of ROS/RNS producing enzymes was analyzed by Western blotting.ResultsDCF fluorescence increased in cells exposed to HEMA for 1 h suggesting the production of hydroxyl radicals, H2O2, or nitric oxide and superoxide anions which form peroxynitrite (ONOO-). Increased DHR123 fluorescence after 24 h indicated the formation of mostly H2O2. The induction of apoptosis in the presence of HEMA was decreased by low concentrations of diphenylene iodonium (DPI), an inhibitor of flavin-containing enzymes. Expression of p47phox, a regulatory subunit of the superoxide producing Nox2, was downregulated, and the expression of NOS which produces nitric oxide (NO) was possibly inhibited by feedback loop mechanisms in HEMA-exposed cultures. Inhibition of HEMA-induced apoptosis by VAS2870 or apocynin further suggested a crucial function of Nox2.SignificanceThe present findings show the physiological relevance of flavin-containing enzymes in monomer-induced oxidative stress and apoptosis.
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影响因子：
--
作者：
Kouros P;Koliniotou-Koumpia E;Koulaouzidou E;Helvatjoglu-Antoniades M;Tziafas D
通讯作者：
Tziafas D
影响因子：
7.4
作者：
Lu, Qing;Wainwright, Mark S.;Harris, Valerie A.;Aggarwal, Saurabh;Hou, Yali;Rau, Thomas;Poulsen, David J.;Black, Stephen M.
通讯作者：
Black, Stephen M.
影响因子：
4.2
作者：
Bouillaguet, S;Wataha, JC;Holz, J
通讯作者：
Holz, J
过氧亚硝酸盐和过氧化氢会引起类似的由 Ccp1 传感器蛋白介导的细胞应激反应。
影响因子：
7.4
作者：
Dorival Martins;Iolie C Bakas;K. McIntosh;A. English
通讯作者：
A. English
DOI：
10.1007/978-1-60761-411-1_4
发表时间：
2010-01-01
期刊：
ADVANCED PROTOCOLS IN OXIDATIVE STRESS II
影响因子：
--
作者：
Eruslanov, Evgeniy;Kusmartsev, Sergei
通讯作者：
Kusmartsev, Sergei