Superoxide production in the vasculature of lipopolysaccharide-treated rats and pigs.
Superoxide production in the vasculature of lipopolysaccharide-treated rats and pigs.
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脂多糖处理的大鼠和猪的脉管系统中超氧化物的产生。
DOI：
10.1097/01.shk.0000054374.88889.37
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发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Magder,SheldonA
中科院分区：
文献类型：
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作者：
Javesghani,Danesh;Hussain,SabahNA;Scheidel,Jonathan;Quinn,MarkT;Magder,SheldonA
脓毒症与活性氧（ROS）的产生增加有关;然而，ROS增加的代谢来源还不清楚。我们推测，最近描述的非吞噬细胞NAD（P）H氧化酶系统可能是脓毒症过程中ROS超氧阴离子（O2-）的重要来源，O2-与一氧化氮（NO）的相互作用可能有助于脓毒症诱导的血管损伤。为了评估这个问题，我们测量了脂多糖（LPS）治疗前后大鼠（诱导型NO合酶产生者（NOSII））和猪（不产生NOSII）的O2 −产生。LPS使大鼠主动脉的O2 −生成量从0.38±0.07 nmol/mg/10 min增加到1.18±0.23 nmol/mg/10 min（P= 0.001），使猪颈动脉的O2 −生成量从0.63±0.05 nmol/mg/10 min增加到1.5±1.6 nmol/mg/10 min（P= 0.001）。在大鼠主动脉和猪主动脉及颈动脉中存在NAD（P）H氧化酶的组分，包括p22 phox、gp 91 phox、p47 phox、p67 phox、mRNA和p22 phox、gp 91 phox蛋白。在大鼠中表达轻度增加，但在猪中未增加。在大鼠中，NADH和NADPH大大增加了O2 −的产生，在未处理的大鼠与LPS处理的大鼠中没有差异。在LPS处理的大鼠中，添加L-NAME使NADH依赖性O 2−的产生从75±3 nmol/O 2−/mg/10 min增加到113±7 nmol/O 2−/mg/10 min，但对未处理的大鼠没有影响。在猪中，即使没有L-NAME，在LPS前和LPS后，NADH刺激的O2 −产生量分别为43±8 nmol/mg/10 min和63±4.3 nmol/mg/10 min（P< 0.05）。与LPS处理的大鼠相比，L-NAME显著降低NADH刺激的O2 −产生（63±4 nmol/mg/10 min至33±5.6 nmol/mg/10 min，P< 0.01）。鲁米诺增强的化学发光也增加了猪颈动脉LPS治疗后，这是一致的过氧亚硝酸盐的形成。我们的研究结果表明，猪和大鼠的血管中存在NAD（P）H氧化酶的成分，并且在LPS后有大量的NADH依赖性O 2−产生增加。然而，NAD（P）H氧化酶在产生NOSII和不产生NOSII的物种中的行为不同，在大鼠中通过NO减少O2 −，而在猪中依赖NO产生。
Sepsis is associated with increased production of reactive oxygen species (ROS); however, the metabolic sources of increased ROS are not well understood. We hypothesized that the recently described nonphagocytic NAD (P) H oxidase system could be an important source of the ROS superoxide anion (O 2−) during sepsis, and the interaction of O 2− with nitric oxide (NO) may contribute to sepsis-induced vascular injury. To evaluate this issue, we measured O 2− production before and after treatment with lipopolysaccharide (LPS) in rats, who are inducible NO synthase producers (NOSII) and in pigs, who do not produce NOSII. LPS increased O 2− production in aorta from rats from 0.38±0.07 nmol/mg/10 min to 1.18±0.23 nmol/mg/10 min,(P= 0.001) in rats, and 0.63±0.05 nmol/mg/10 min to 1.5±1.6 nmol/mg/10 min (P= 0.001) in carotid arteries from pigs. Components of NAD (P) H oxidase, including p22 phox, gp91 phox, p47 phox, p67 phox, mRNA and p22 phox, and gp91 phox proteins were present in rat aorta and aorta and carotid arteries from pigs. Expression mildly increased in rats, but not in pigs. In rats, NADH and NADPH greatly increased O 2− production with no difference in untreated versus LPS-treated rats. The addition of L-NAME increased NADH-dependant O 2− production from 75±3 nmol/O 2−/mg/10 min to 113±7 nmol/O 2−/mg/10 min in LPS-treated rats, but had no effect in untreated rats. In pigs, the NADH-stimulated O 2− production was 43±8 nmol/mg/10 min before and 63±4.3 nmol/mg/10 min after LPS even without L-NAME (P< 0.05). In contrast to LPS-treated rats, L-NAME markedly decreased NADH-stimulated O 2− production (63±4 nmol/mg/10 min to 33±5.6 nmol/mg/10 min, P< 0.01). Luminol-enhanced chemiluminescence was also increased in porcine carotid arteries after LPS treatment, which is consistent with peroxynitrite formation. Our results indicate that components of NAD (P) H oxidase are present in vessels of pigs and rats and there is substantial NADH-dependent O 2− production that is increased after LPS. However, the behavior of NAD (P) H oxidase in NOSII-producing and nonproducing species differs with a reduction of O 2− by NO in rats and NO-dependent production in pigs.