Department of Medicine, University of Wisconsin, Madison, USA.
Am J Physiol Heart Circ Physiol. 2010 Dec;299(6):H1884-90. doi: 10.1152/ajpheart.00363.2010. Epub 2010 Oct 8.
The sulfonylurea receptor-2 (SUR2) is a subunit of ATP-sensitive potassium channels (K(ATP)) in heart. Mice with the SUR2 gene disrupted (SUR2m) are constitutively protected from ischemia-reperfusion (I/R) cardiac injury. This was surprising because K(ATP), either sarcolemmal or mitochondrial or both, are thought to be important for cardioprotection. We hypothesized that SUR2m mice have an altered mitochondrial phenotype that protects against I/R. Mitochondrial membrane potential (ΔΨ(m)), tolerance to Ca(2+) load, and reactive oxygen species (ROS) generation were studied by fluorescence-based assays, and volumetric changes in response to K(+) were measured by light scattering in isolated mitochondria. For resting SUR2m mitochondria compared with wild type, the ΔΨ(m) was less polarized (46.1 ± 0.4 vs. 51.9 ± 0.6%), tolerance to Ca(2+) loading was increased (163 ± 2 vs. 116 ± 2 μM), and ROS generation was enhanced with complex I [8.5 ± 1.2 vs. 4.9 ± 0.2 arbitrary fluorescence units (afu)/s] or complex II (351 ± 51.3 vs. 166 ± 36.2 afu/s) substrates. SUR2m mitochondria had greater swelling in K(+) medium (30.2 ± 3.1%) compared with wild type (14.5 ± 0.6%), indicating greater K(+) influx. Additionally, ΔΨ(m) decreased and swelling increased in the absence of ATP in SUR2m, but the sensitivity to ATP was less compared with wild type. When the mitochondria were subjected to hypoxia-reoxygenation, the decrease in respiration rates and respiratory control index was less in SUR2m. ΔΨ(m) maintenance in the SUR2m intact myocytes was also more tolerant to metabolic inhibition. In conclusion, the cardioprotection observed in the SUR2m mice is associated with a protected mitochondrial phenotype resulting from enhanced K(+) conductance that partially dissipated ΔΨ(m). These results have implications for possible SUR2 participation in mitochondrial K(ATP).
磺酰脲受体-2(SUR2)是心脏中 ATP 敏感性钾通道(K(ATP))的亚基。SUR2 基因缺失(SUR2m)的小鼠对缺血再灌注(I/R)心脏损伤具有持续的保护作用。这令人惊讶,因为 K(ATP),无论是肌浆网或线粒体或两者,都被认为对心脏保护很重要。我们假设 SUR2m 小鼠具有改变的线粒体表型,可防止 I/R。通过荧光测定法研究线粒体膜电位(ΔΨ(m))、对 Ca(2+)负荷的耐受性和活性氧(ROS)的产生,通过光散射测量对 K(+)的体积变化。与野生型相比,在静止的 SUR2m 线粒体中,ΔΨ(m)的极化程度较低(46.1 ± 0.4 对 51.9 ± 0.6%),对 Ca(2+)加载的耐受性增加(163 ± 2 对 116 ± 2 μM),并且在存在复合物 I [8.5 ± 1.2 对 4.9 ± 0.2 任意荧光单位(afu)/s]或复合物 II(351 ± 51.3 对 166 ± 36.2 afu/s)底物时,ROS 的产生增强。与野生型相比,SUR2m 线粒体在 K(+)介质中的肿胀更大(30.2 ± 3.1%),表明 K(+)流入增加。此外,在 SUR2m 中没有 ATP 时,ΔΨ(m)减少并且肿胀增加,但是与野生型相比,对 ATP 的敏感性降低。当线粒体受到缺氧-复氧时,SUR2m 中的呼吸速率和呼吸控制指数的下降较小。在 SUR2m 完整心肌细胞中,ΔΨ(m)的维持对代谢抑制也更耐受。总之,在 SUR2m 小鼠中观察到的心脏保护作用与增强的 K(+)电导相关联,该增强的 K(+)电导部分耗散了 ΔΨ(m)。这些结果表明 SUR2 可能参与线粒体 K(ATP)。