Shetewy Aza, Shimada-Takaura Kayoko, Warner Danielle, Jong Chian Ju, Mehdi Abu-Bakr Al, Alexeyev Mikhail, Takahashi Kyoko, Schaffer Stephen W
Department of Pharmacology, University of South Alabama College of Medicine, Mobile, AL, 36688, USA.
Department of Cell Biology/Neuroscience, University of South Alabama College of Medicine, Mobile, USA.
Mol Cell Biochem. 2016 May;416(1-2):11-22. doi: 10.1007/s11010-016-2688-z. Epub 2016 Mar 29.
Hyper-beta-alaninemia is a rare metabolic condition that results in elevated plasma and urinary β-alanine levels and is characterized by neurotoxicity, hypotonia, and respiratory distress. It has been proposed that at least some of the symptoms are caused by oxidative stress; however, only limited information is available on the mechanism of reactive oxygen species generation. The present study examines the hypothesis that β-alanine reduces cellular levels of taurine, which are required for normal respiratory chain function; cellular taurine depletion is known to reduce respiratory function and elevate mitochondrial superoxide generation. To test the taurine hypothesis, isolated neonatal rat cardiomyocytes and mouse embryonic fibroblasts were incubated with medium lacking or containing β-alanine. β-alanine treatment led to mitochondrial superoxide accumulation in conjunction with a decrease in oxygen consumption. The defect in β-alanine-mediated respiratory function was detected in permeabilized cells exposed to glutamate/malate but not in cells utilizing succinate, suggesting that β-alanine leads to impaired complex I activity. Taurine treatment limited mitochondrial superoxide generation, supporting a role for taurine in maintaining complex I activity. Also affected by taurine is mitochondrial morphology, as β-alanine-treated fibroblasts undergo fragmentation, a sign of unhealthy mitochondria that is reversed by taurine treatment. If left unaltered, β-alanine-treated fibroblasts also undergo mitochondrial apoptosis, as evidenced by activation of caspases 3 and 9 and the initiation of the mitochondrial permeability transition. Together, these data show that β-alanine mediates changes that reduce ATP generation and enhance oxidative stress, factors that contribute to heart failure.
高β-丙氨酸血症是一种罕见的代谢疾病,会导致血浆和尿液中β-丙氨酸水平升高,其特征为神经毒性、肌张力减退和呼吸窘迫。有人提出,至少部分症状是由氧化应激引起的;然而,关于活性氧生成机制的信息有限。本研究检验了以下假设:β-丙氨酸会降低细胞内牛磺酸水平,而正常呼吸链功能需要牛磺酸;已知细胞内牛磺酸耗竭会降低呼吸功能并增加线粒体超氧化物生成。为了验证牛磺酸假说,将分离出的新生大鼠心肌细胞和小鼠胚胎成纤维细胞分别在不含或含有β-丙氨酸的培养基中培养。β-丙氨酸处理导致线粒体超氧化物积累,同时氧消耗减少。在暴露于谷氨酸/苹果酸的透化细胞中检测到β-丙氨酸介导的呼吸功能缺陷,但在利用琥珀酸的细胞中未检测到,这表明β-丙氨酸会导致复合体I活性受损。牛磺酸处理限制了线粒体超氧化物生成,支持了牛磺酸在维持复合体I活性中的作用。线粒体形态也受牛磺酸影响,因为经β-丙氨酸处理的成纤维细胞会发生碎片化,这是线粒体不健康的迹象,而牛磺酸处理可使其恢复。如果不改变,经β-丙氨酸处理的成纤维细胞也会发生线粒体凋亡,这可通过半胱天冬酶3和9的激活以及线粒体通透性转换的启动得到证明。总之,这些数据表明,β-丙氨酸介导的变化会减少ATP生成并增强氧化应激,这些因素会导致心力衰竭。