Hydrogen peroxide induces apoptosis in human dental pulp cells via caspase-9 dependent pathway.
Hydrogen peroxide induces apoptosis in human dental pulp cells via caspase-9 dependent pathway.
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DOI：
10.1016/j.joen.2013.06.006
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发表时间：
2013-09
影响因子：
4.2
通讯作者：
Tian-tian Wu;Li-fen Li-Li-fen-Li-47681337;R. Du;Long Jiang;Ya-qin Zhu
中科院分区：
文献类型：
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作者：
Tian-tian Wu;Li-fen Li-Li-fen-Li-47681337;R. Du;Long Jiang;Ya-qin Zhu
前言活性氧是真核细胞氧化代谢过程中产生的一组代谢中间产物。它们包括超氧阴离子(O2−)、过氧化氢(H2O2)、羟基自由基(·OH)和1O2。在这些中间体中，过氧化氢是最稳定的。牙髓细胞可能受到牙齿漂白、激光辐射和牙科材料的侵袭。这会影响细胞内的活性氧水平。细胞凋亡是最常见的程序性细胞死亡形式，对组织发育和再生至关重要。关于H_2O_2与人牙髓细胞(HDPC)凋亡之间的关系的研究很少。方法采用改良的组织块体外培养技术，在37℃、20%O2(5%CO2、95%空气)的改良Eagle培养基中培养hDPC。用四甲基偶氮唑蓝比色法检测细胞存活率。应用Annexin V-异硫氰酸荧光素/碘化丙啶细胞凋亡法和流式细胞术检测细胞凋亡。结果培养1~7天的HDPC细胞存活率较对照组明显下降。20和50μ/L双氧水作用于HDPC后，细胞相对凋亡数及活化Caspase-3和裂解Caspase-9的表达均显著增加。结论低浓度的过氧化氢对HDPC具有细胞毒作用，并以caspase-9依赖的方式诱导HDPC凋亡。
IntroductionReactive oxygen species are a group of metabolic intermediates produced during oxidative metabolism in eukaryotic cells. They include superoxide anion (O2−), hydrogen peroxide (H2O2), hydroxyl radical (·OH), and1O2. Of these intermediates, H2O2is the most stable. Dental pulp cells can be invaded by tooth bleaching, laser radiation, and dental materials. This can influence the intracellular level of reactive oxygen species. Apoptosis, which is the best-known form of programmed cell death, is pivotal to tissue development and regeneration. Little information is available regarding the relationship between H2O2and apoptosis of human dental pulp cells (hDPCs). The purpose of this study was to investigate whether H2O2can induce apoptosis in hDPCs and its signaling way.MethodsHDPCs were obtained by using a modified tissue explant techniquein vitroand cultured at 37°C, 20% O2(5% CO2, 95% air) in Dulbecco modified Eagle medium. Cell viability was investigated by methyl-thiazol-tetrazolium assay. Cell apoptosis was detected by using the annexin V–fluorescein isothiocyanate/propidium iodide apoptosis assay and flow cytometry. Expression of activated caspase-3, cleaved caspase-9, and β-actin was analyzed by using Western blot.ResultsCell viability of hDPCs decreased more in treated groups than in the control group from days 1 to 7. The relative number of apoptotic cells and the expression of activated caspase-3 and cleaved caspase-9 were much higher in groups exposed to 20 and 50 μmol/L H2O2.ConclusionsThese results imply that low concentrations of H2O2are cytotoxic to hDPCs and induce apoptosis in hDPCs in a caspase-9–dependent way.