Boveris Alberto, Valdez Laura B, Alvarez Silvia, Zaobornyj Tamara, Boveris Alejandro D, Navarro Ana
Laboratory of Free Radical Biology, School of Pharmacy and Biochemistry, University of Buenos Aires, Buenos Aires, Argentina.
Antioxid Redox Signal. 2003 Jun;5(3):265-71. doi: 10.1089/152308603322110841.
Nitric oxide synthase activity was recognized in rat renal cortex mitochondria (mtNOS) with nitric oxide (NO) production rates of 0.14-0.78 nmol/min/mg of protein. Rat pretreatment with enalapril (30 mg/kg/day i.p., up to 15 days) increased NO production in kidney, liver, and heart mitochondria. In kidney, mtNOS activity and mtNOS protein, measured by western blot densitometry, were 5 and 2.3 times increased, respectively. Electron paramagnetic resonance analysis with the probe N-methyl-D-glucamine dithiocarbamate/FeSO(4) detected NO production in mitochondria isolated from enalapril-treated rats, but not in control untreated animals. Polyclonal antibodies anti-iNOS and anti-nNOS detected kidney mtNOS in western blots and inhibited mtNOS biochemical activity. The enzymatic activity of kidney mtNOS generates intramitochondrial NO concentrations that regulate mitochondrial functions: state 3 respiration was decreased by 12-28%, and state 4 hydrogen peroxide production was increased 12-35%.
在大鼠肾皮质线粒体中发现了一氧化氮合酶活性（mtNOS），一氧化氮（NO）生成速率为0.14 - 0.78 nmol/分钟/毫克蛋白质。用依那普利（腹腔注射30毫克/千克/天，最多15天）预处理大鼠可增加肾脏、肝脏和心脏线粒体中的NO生成。在肾脏中，通过蛋白质免疫印迹密度测定法测得的mtNOS活性和mtNOS蛋白分别增加了5倍和2.3倍。用探针N - 甲基 - D - 葡糖胺二硫代氨基甲酸盐/硫酸亚铁（N-methyl-D-glucamine dithiocarbamate/FeSO₄）进行电子顺磁共振分析，在从依那普利处理的大鼠分离的线粒体中检测到了NO生成，但在未处理的对照动物中未检测到。抗诱导型一氧化氮合酶（anti-iNOS）和抗神经元型一氧化氮合酶（anti-nNOS）的多克隆抗体在蛋白质免疫印迹中检测到肾脏mtNOS，并抑制mtNOS的生化活性。肾脏mtNOS的酶活性产生线粒体内的NO浓度，该浓度调节线粒体功能：状态3呼吸降低了12 - 28%，状态4过氧化氢生成增加了12 - 35%。
