High glucose-induced apoptosis in human endothelial cells is mediated by sequential activations of c-Jun NH2-terminal kinase and caspase-3
High glucose-induced apoptosis in human endothelial cells is mediated by sequential activations of c-Jun NH2-terminal kinase and caspase-3
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DOI：
10.1161/01.cir.101.22.2618
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发表时间：
2000-06-06
期刊：
影响因子：
37.8
通讯作者：
Lin-Shiau, SY
中科院分区：
文献类型：
--
作者：
Ho, FM;Liu, SH;Lin-Shiau, SY
背景-糖尿病引起多种心血管并发症。高糖可诱导内皮细胞产生活性氧和凋亡。高糖诱导内皮细胞凋亡的分子机制目前知之甚少。方法与结果-我们阐明了高糖诱导人脐静脉内皮细胞（HUVECs）凋亡的信号通路。在存在或不存在抗氧化剂抗坏血酸的情况下，用含有5.5、19或33 mmol/L葡萄糖的培养基处理HUVEC。流式细胞仪检测细胞内H2 O2水平。细胞凋亡检测采用ELISA法和形态学Hoechst染色法。高糖能够诱导c-Jun氨基末端激酶（JNK）的活性，但不能诱导细胞外信号调节激酶1/2或p38丝裂原活化蛋白激酶的活性。此外，我们发现白细胞介素1 β转换酶（ICE）/CED-3家族蛋白酶（caspase-3）在高糖诱导的细胞凋亡中被激活。Caspase-3/CPP 32特异性抑制剂Ac-DEVD-CHO可抑制高糖诱导的细胞凋亡。JNK 1反义寡核苷酸能抑制caspase-3活性，但不影响H_2O_2的产生，并能阻断高糖诱导的细胞凋亡。此外，H2 O2的产生，JNK的活性，caspase-3的活性，以及随后的凋亡诱导高糖可抑制抗坏血酸conclusion-The本研究表明，活性氧诱导高糖可能参与JNK激活，这反过来又触发caspase-3，促进HUVECs的凋亡。
Background-Diabetes mellitus causes multiple cardiovascular complications. High glucose can induce reactive oxygen species and apoptosis in endothelial cells. Little is known about the molecular mechanisms in high glucose-induced endothelial cell apoptosis.Methods and Results-We elucidated the signaling pathway of high glucose-induced apoptosis in human umbilical vein endothelial cells (HUVECs). HUVECs were treated with media containing 5.5, 19, or 33 mmol/L of glucose in the presence or absence of an antioxidant, ascorbic acid. The level of intracellular H2O2 was measured by flow cytometry. For detection of apoptosis, the cell death detection ELISA assay and the morphological Hoechst staining were used. High glucose was capable of inducing the activity of c-Jun NH2-terminal kinase (JNK) but not extracellular signal-regulated kinase 1/2 or p38 mitogen-activated protein kinase during the treatment periods, as evidenced by immunocomplex kinase assay. Moreover, we found that the interleukin 1 beta-converting enzyme (ICE)/CED-3 family protease (caspase-3) became activated in high glucose-induced apoptosis. Caspase-3/CPP32-specific inhibitor, Ac-DEVD-CHO, could inhibit high glucose-induced apoptosis. Furthermore, we found that JNK1 specific antisense oligonucleotide could suppress caspase-3 activity but not affect H2O2 generation and could block apoptosis induced by high glucose. Also, H2O2 generation, JNK activity, caspase-3 activity, and the subsequent apoptosis induced by high glucose could be suppressed by ascorbic acid.Conclusions-The present study indicates that reactive oxygen species induced by high glucose may be involved in JNK activation, which in turn triggers the caspase-3 that facilitates the apoptosis in HUVECs.