Hine Christopher, Mitchell James R
Department of Cellular and Molecular Medicine, Cleveland Clinic Lerner Research Institute, Cleveland, OH 44195 USA.
Department of Genetics and Complex Diseases, Harvard T.H. Chan School of Public Health, Boston, MA 02115 USA.
Bio Protoc. 2017 Jul 5;7(13). doi: 10.21769/BioProtoc.2382.
Hydrogen sulfide (HS) gas is produced in cells and tissues via various enzymatic processes. HS is an important signaling molecule in numerous biological processes, and deficiencies in endogenous HS production are linked to cardiovascular and other health complications. Quantitation of steady-state HS levels is challenging due to volatility of the gas and the need for specialized equipment. However, the capacity of an organ or tissue extract to produce HS under optimized reaction conditions can be measured by a number of current assays that vary in sensitivity, specificity and throughput capacity. We developed a rapid, inexpensive, specific and relatively high-throughput method for quantitative detection of HS production capacity from biological tissues. HS released into the head space above a biological sample reacts with lead acetate to form lead sulfide, which is measured on a continuous basis using a plate reader or as an endpoint assay.
硫化氢(HS)气体通过各种酶促过程在细胞和组织中产生。HS是众多生物过程中的重要信号分子,内源性HS生成不足与心血管及其他健康并发症有关。由于该气体的挥发性以及对专业设备的需求,对稳态HS水平进行定量分析具有挑战性。然而,在优化的反应条件下,器官或组织提取物产生HS的能力可以通过多种当前的检测方法来测量,这些方法在灵敏度、特异性和通量能力方面存在差异。我们开发了一种快速、廉价、特异且通量相对较高的方法,用于定量检测生物组织产生HS的能力。释放到生物样品上方顶空中的HS与醋酸铅反应形成硫化铅,可使用酶标仪连续测量或作为终点检测法进行测量。