Hydrogen peroxide induces apoptosis of chondrocytes; involvement of calcium ion and extracellular signal-regulated protein kinase
Hydrogen peroxide induces apoptosis of chondrocytes; involvement of calcium ion and extracellular signal-regulated protein kinase
复制标题
DOI：
10.1007/s000110050719
复制
发表时间：
2001-01-01
影响因子：
6.7
通讯作者：
Hamanisi, C
中科院分区：
文献类型：
--
作者：
Asada, S;Fukuda, K;Hamanisi, C
目的：最近的研究表明活性氧促进软骨的降解。我们证明过氧化氢（H2 O2）会抑制软骨细胞的蛋白聚糖合成、诱导细胞凋亡并刺激细胞外信号调节蛋白激酶（ERK）（Inflamm Res 48. 399-403，1999）。为了确定ERK的激活是否参与软骨细胞凋亡的诱导，我们研究了在此过氧化氢诱导apoptosis.Design的信号转导通路：牛关节软骨细胞进行培养。为了确定凋亡的诱导，使用膜联蛋白V染色和末端脱氧核苷酸转移酶。使用apopain测定试剂盒测量caspase-3的活性。结果：过氧化氢可增强Annexin V阳性凋亡细胞和凋亡执行者caspase-3的活性。过氧化氢也增强了细胞内钙离子和预孵育与细胞内钙离子螯合剂保护软骨细胞对过氧化氢诱导的细胞凋亡，这表明，在胞浆钙离子的增加在这一行动中起着决定性的作用。结论：过氧化氢通过Ca ~（2+）信号途径诱导软骨细胞凋亡，ERK参与了这些信号转导途径。
Objective: Recent observations demonstrated that reactive oxygen species facilitate cartilage degradation. We demonstrated that hydrogen peroxide (H2O2) caused inhibition of proteoglycan synthesis, induction of apoptosis and stimulation of extracellular signal-regulated protein kinase (ERK) of the chondrocytes (Inflamm Res 48. 399-403, 1999). To determine whether activation of ERK is involved in the induction of chondrocyte apoptosis, we examined the signal transduction pathways in this hydrogen peroxide induced apoptosis.Design: Bovine articular chondrocytes were cultured. To determine the induction of apoptosis, Annexin V staining and terminal deoxynucleotidyl transferase were used. The activity of caspase-3 was measured using an apopain assay kit. Intracellular Ca2+ imaging was observed after fura2-AM loading.Results: Hydrogen peroxide enhanced annexin V positive apoptotic cells and caspase-3 activity, which is an executor of apoptosis. Hydrogen peroxide also enhanced intracellular Ca2+ and preincubation with the intracellular Ca2+ chelator protected chondrocytes against hydrogen peroxide-induced cell apoptosis, indicating that an increase in the cytosolic Ca2+ plays a decisive role in this action. When ERK activity was blocked with geldanamycin and PD098059, increased apoptosis was evident.Conclusion: Hydrogen peroxide induces chondrocyte apoptosis via Ca2+ signaling, and ERK is involved in these signal transduction pathways.