Hyperosmotic Stress Induces Cell Death in an Odontoblast-lineage Cell Line
Hyperosmotic Stress Induces Cell Death in an Odontoblast-lineage Cell Line
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DOI：
10.1016/j.joen.2012.03.023
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发表时间：
2012-07-01
影响因子：
4.2
通讯作者：
Torii, Mitsuo
中科院分区：
文献类型：
--
作者：
Fujisawa, Mari;Tokuda, Masayuki;Torii, Mitsuo
前言：渗透应力是与龋齿或牙本质过敏引起的牙痛有关的刺激之一。牙周炎引起的牙痛的机制还不完全清楚。本研究旨在探讨成牙本质细胞对蔗糖诱导的高渗应激的反应。方法：我们在实验中使用成牙本质细胞系（OLC）。用蔗糖刺激OLC产生高渗应激。采用逆转录聚合酶链反应（RT-PCR）检测牙本质涎磷蛋白（DSPP）和牙本质基质蛋白1（DSP 1）的表达。采用3-（4，5-二甲基噻唑-2-基）-2，5-二苯基四氮唑测定法检测OLC的细胞活力。采用4-6-diamidino-2-phenylindole染色、caspase-3蛋白印迹和annexin V检测细胞死亡。Western blot法检测丝裂原活化蛋白激酶（MAPK）的表达。结果：高渗应激对OLC中DSPP和DSP 1的表达无影响。细胞培养3小时，细胞活力下降超过700 mOsm。在高渗胁迫下，细胞和细胞核的形状变得不规则和空泡化。高渗应激处理后，caspase-3的表达增加。流式细胞仪分析中检测到一些碘化丙啶阳性细胞。高渗胁迫可诱导3个MAPK磷酸化。在500和700 mOsm下，3种MAPK的抑制剂抑制高渗应激诱导的细胞活力下降。结论：高渗应激通过MAPK途径诱导OLC细胞死亡。（J Endod 2012;38：931-935）
Introduction: Osmotic stress is one of the stimulations related to dental pain caused by caries or dentin hypersensitivity. The mechanism of osmotic-induced dental pain is not completely understood. The purpose of this study was to examine the responses of odontoblasts under sucrose-induced hyperosmotic stress. Methods: We used an odontoblast-lineage cell (OLC) line in our experiments. OLCs were stimulated with sucrose to produce hyperosmotic stress. The expressions of dentin sialophosphoprotein (DSPP) and dentin matrix protein 1 (DMP 1) were detected by using reverse transcriptase polymerase chain reaction assay. The cell viability of OLCs was detected by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium assay. The responses accompanied with cell death were detected by using 4-6-diamidino-2-phenylindole staining, Western blotting of caspase-3, and annexin V assay. The expression of mitogen-activated protein kinases (MAPKs) was detected by using Western blot analysis. Results: DSPP and DMP 1 were not affected by hyperosmotic stress in OLCs. Cell viability decreased over 700 mOsm for 3 hours of cell culture. The shapes of cells and nuclei became irregular and vacuolar under hyperosmotic stress. The expression of cleaved caspase-3 was increased after treatment with hyperosmotic stress. Some propidium iodide-positive cells were detected in flow cytometry analysis. Phosphorylation of 3 MAPKs was induced by hyperosmotic stress. Inhibitors of 3 MAPKs inhibited the hyperosmotic stress-induced decline in cell viability at 500 and 700 mOsm. Conclusions: Hyperosmotic stress induces cell death of OLCs with sucrose through a MAPK pathway. (J Endod 2012;38:931-935)