Involvement of anion exchanger-2 in apoptosis of endothelial cells induced by high glucose through an mPTP-ROS-Caspase-3 dependent pathway
Involvement of anion exchanger-2 in apoptosis of endothelial cells induced by high glucose through an mPTP-ROS-Caspase-3 dependent pathway
复制标题
阴离子交换剂2通过mPTP-ROS-Caspase-3依赖性途径参与高糖诱导的内皮细胞凋亡
DOI：
10.1007/s10495-010-0477-9
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发表时间：
2010-06-01
期刊：
影响因子：
7.2
通讯作者：
He, Ming
中科院分区：
文献类型：
--
作者：
Huang, Qi-Ren;Li, Qing;He, Ming
内皮细胞(EC)的过度凋亡在糖尿病血管病变的发生发展中起重要作用。阴离子交换因子-2(AE2)可能参与了血管病变。然而，AE2介导内皮细胞凋亡的分子机制尚不清楚。本研究旨在探讨AE2在高糖诱导人脐静脉内皮细胞凋亡中的作用及其可能机制。首先，在不同的时间点(分别为12、24、48、72、120和168h)，将人脐静脉内皮细胞暴露于不同的葡萄糖浓度(分别为5.5、17.8、35.6、71.2和142.4 mmol/L，pH=77.40)中。检测细胞内Cl-−浓度([Cl-−]i)、AE2表达及细胞凋亡率。然后，用4，4‘-二异硫氰基二苯乙烯-2，2’-二磺酸(DIDS)、无氯−培养液或Ae2的特异性RNA干扰(RNAi)来确定Ae2是否能介导HG诱导的细胞凋亡。最后，通过检测线粒体通透性转换孔(MPTP、ΔΨm)开口量、活性氧(ROS)水平和Caspase3活性，探讨了Ae2诱导细胞凋亡的机制。我们发现，HG以时间和浓度依赖的方式上调AE2的表达和活性，升高[Cl−]i，诱导细胞凋亡。AE2可能通过MPTP-ROS-Caspase-3依赖途径介导HUVECs的凋亡。这些发现提示AE2可能是一种葡萄糖敏感的跨膜转运蛋白，是治疗糖尿病血管病变的一个新的潜在靶点。
Excess apoptosis of endothelial cells (EC) plays crucial roles in the onset and progression of vasculopathy in diabetes mellitus. Anion exchanger-2 (AE2) might be involved in the vasculopathy. However, little is known about the molecular mechanisms that AE2 mediated the apoptosis of EC. The purpose of this study was to explore the role of AE2 in the apoptosis of HUVECs induced by high glucose (HG) and its possible mechanisms. First, HUVECs were exposed to different glucose concentrations (5.5, 17.8, 35.6, 71.2 and 142.4 mmol/l, respectively, pH = 7.40) for different time points (12, 24, 48, 72, 120, and 168 h, respectively). Intracellular Cl−concentration ([Cl−]i), AE2 expression and the apoptosis were assayed. Then, 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS), Cl−-free media or specific RNA interference (RNAi) for AE2 was used to confirm whether AE2 could mediate the apoptosis induced by HG. Finally, the mechanisms of the AE2-mediated apoptosis were investigated by detecting mitochondrial permeability transition pore (mPTP, ΔΨm) openings, reactive oxygen species (ROS) levels and Caspase-3 activity. We found that HG upregulated the AE2 expression and activity, increased [Cl−]i and induced the apoptosis in a time- and concentration-dependent manner. The apoptosis of HUVECs by HG was possibly mediated by AE2 through an mPTP-ROS-Caspase-3 dependent pathway. These findings suggested that AE2 was likely to be a glucose-sensitive transmembrane transporter and a novel potential therapeutic target for diabetic vasculopathy.