Anion Exchanger 3 Is Required for Sasanquasaponin to Inhibit Ischemia/Reperfusion-Induced Elevation of Intracellular Cl- Concentration and to Elicit Cardioprotection
Anion Exchanger 3 Is Required for Sasanquasaponin to Inhibit Ischemia/Reperfusion-Induced Elevation of Intracellular Cl- Concentration and to Elicit Cardioprotection
复制标题
阴离子交换剂 3 是油茶皂苷抑制缺血/再灌注引起的细胞内 Cl- 浓度升高并引起心脏保护作用所必需的
DOI：
10.1002/jcb.23195
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发表时间：
2011-10-01
影响因子：
4
通讯作者：
Huang, Min
中科院分区：
文献类型：
--
作者：
Chen, He-Ping;He, Ming;Huang, Min
近年来研究表明，油茶皂苷（SQS）对缺血再灌注损伤的心脏保护作用与抑制缺血再灌注引起的细胞内Cl-浓度（[Cl-]（i））升高有关。然而，抑制机制仍不清楚。阴离子交换蛋白3（AE（3））是[Cl-]（i）的重要调节蛋白。本研究探讨AE（3）是否在SQS抑制缺血/再灌注引起的[Cl-]（i）升高的作用中起关键作用，并介导SQS对H9 c2细胞的心脏保护作用。将正常和AE（3）-敲低的H9 c2细胞与或不与不同浓度的SQS（0.1、1或10 μ M）一起孵育24小时，随后进行模拟缺血/再灌注（sI/R）。Western blot检测AE（3）的表达。采用流式细胞仪分析测定[Cl-]（i）、[Ca 2 +]（i）、活性氧（ROS）产生和细胞凋亡。结果表明，SQS预处理浓度依赖性地减弱sI/R诱导的正常H9 c2细胞活力丧失和乳酸脱氢酶渗漏。另外，SQS浓度依赖性地上调AE（3）蛋白表达，抑制sI/R诱导的[Cl-]（i）升高，降低Ca 2+超载、ROS产生和细胞凋亡。然而，SQS对AE（3）基因敲低的H9 c2细胞的剂量依赖性心肌保护作用被取消，SQS对[Cl-]（i）、Ca 2+超载、ROS产生和细胞凋亡的抑制作用也被逆转。我们的数据表明，AE（3）介导SQS对sI/R损伤的心脏保护作用。重要的是，SQS需要AE（3）来抑制sI/R诱导的[Cl-]（i）升高，[Cl-]（i）随后抑制sI/R诱导的Ca 2+过载、ROS产生和细胞凋亡。J.细胞。112：2803-2812，2011. (C)2011 Wiley-Liss，Inc.
Recent studies have shown that the cardioprotection of sasanquasaponin (SQS) against ischemia/reperfusion injury is related to inhibiting ischemia/reperfusion-induced elevation of intracellular Cl- concentration ([Cl-](i)). However, the mechanism of inhibition remains unclear. Anion exchanger 3 (AE(3)) is an important regulatory protein for [Cl-](i). This study investigated whether AE(3) plays the critical role in the inhibitory effect of SQS on elevation of [Cl-](i) induced by ischemia/reperfusion and mediates the cardioprotection of SQS in H9c2 cells. Normal and AE(3)-knockdown H9c2 cells were incubated for 24 h with or without various concentrations of SQS (0.1, 1, or 10 mu M) followed by simulated ischemia/reperfusion (sI/R). AE(3) expression was detected by Western blot. Flow cytometer analysis was employed to determine [Cl-](i), [Ca2+](i), reactive oxygen species (ROS) production, and cell apoptosis. The results showed that SQS pretreatment concentration-dependently attenuated sI/R-induced viability loss and lactate dehydrogenase leakage in normal H9c2 cells. Additionally, SQS concentration-dependently up-regulated AE(3) protein expression, and inhibited sI/R-induced the elevation of [Cl-](i) followed by the attenuation of Ca2+ overload, ROS production, and cell apoptosis. However, the dose-dependent cardioprotection induced by SQS was abolished in AE(3)-knockdown H9c2 cells, and the inhibitory effects of SQS on [Cl-](i), Ca2+ overload, ROS production, and cell apoptosis were also reversed. Our data indicate that AE(3) mediates the cardioprotective effect of SQS against sI/R injury. Importantly, AE(3) is required for SQS to inhibit sI/R-induced elevation of [Cl-](i), which subsequently inhibited sI/R-induced Ca2+ overload, ROS production, and cell apoptosis. J. Cell. Biochem. 112: 2803-2812, 2011. (C) 2011 Wiley-Liss, Inc.