Owiredu Shawn, Ranganathan Abhay, Greenwood John C, Piel Sarah, Janowska Joanna I, Eckmann David M, Kelly Matthew, Ehinger Johannes K, Kilbaugh Todd J, Jang David H
Department of Emergency Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, United States.
Children's Hospital of Philadelphia (CHOP), Philadelphia, PA, 19104, United States.
Toxicol Rep. 2020 Sep 17;7:1263-1271. doi: 10.1016/j.toxrep.2020.09.002. eCollection 2020.
The objective of this study was to compare the use of hydroxocobalamin (B12a) and a succinate prodrug to evaluate for improvement in mitochondrial function in an model of cyanide poisoning. Peripheral blood mononuclear cells (PBMC) and human aortic smooth muscle cells (HASMC) incubated with 50 mM of sodium cyanide (CN) for five minutes serving as the CN group compared to controls. We investigated the following: (1) Mitochondrial respiration; (2) Superoxide and mitochondrial membrane potential with microscopy; (3) Citrate synthase protein expression. All experiments were performed with a cell concentration of 2-3 × 10 cells/ml for both PBMC and HASMC. There were four conditions: (1) Control (no exposure); (2) Cyanide (exposure only); (3) B12a (cyanide exposure followed by B12a treatment); (4) NV118 (cyanide followed by NV118 treatment). In this study the key findings include: (1) Improvement in key mitochondrial respiratory states with the succinate prodrug (NV118) but not B12a; (2) Attenuation of superoxide production with treatment of NV118 but not with B12a treatment; (3) The changes in respiration were not secondary to increased mitochondrial content as measured by citrate synthase; (4) The use of easily accessible human blood cells showed similar mitochondrial response to both cyanide and treatment to HASMC. The use of a succinate prodrug to circumvent partial CIV inhibition by cyanide with clear reversal of cellular respiration and superoxide production that was not attributed to changes in mitochondrial content not seen by the use of B12a.
本研究的目的是比较使用羟钴胺素（B12a）和一种琥珀酸酯前药，以评估在氰化物中毒模型中线粒体功能的改善情况。与对照组相比，外周血单个核细胞（PBMC）和人主动脉平滑肌细胞（HASMC）用50 mM氰化钠（CN）孵育5分钟作为CN组。我们研究了以下内容：（1）线粒体呼吸；（2）通过显微镜观察超氧化物和线粒体膜电位；（3）柠檬酸合酶蛋白表达。所有实验均在PBMC和HASMC细胞浓度为2-3×10个细胞/ml的条件下进行。有四种情况：（1）对照组（无暴露）；（2）氰化物组（仅暴露）；（3）B12a组（氰化物暴露后用B12a治疗）；（4）NV118组（氰化物暴露后用NV118治疗）。在本研究中，主要发现包括：（1）琥珀酸酯前药（NV118）可改善关键的线粒体呼吸状态，但B12a不能；（2）NV118治疗可减轻超氧化物的产生，但B12a治疗不能；（3）呼吸变化并非由柠檬酸合酶测量的线粒体含量增加所致；（4）使用易于获取的人类血细胞显示，对氰化物和治疗的线粒体反应与HASMC相似。使用琥珀酸酯前药可规避氰化物对细胞色素c氧化酶（CIV）的部分抑制，明显逆转细胞呼吸和超氧化物的产生，这并非归因于线粒体含量的变化，而使用B12a则未观察到这种变化。
