Kukreja R C, Jesse R L, Hess M L
Department of Medicine, Medical College of Virginia, Richmond 23298.
Mol Cell Biochem. 1992 Apr;111(1-2):17-24. doi: 10.1007/BF00229569.
The purpose of this study was to explore the role of singlet oxygen in cardiovascular injury. To accomplish this objective, we investigated the effect of singlet oxygen [generated from photoactivation of rose-bengal] on the calcium transport and Ca(2+)-ATPase activity of cardiac sarcoplasmic reticulum and compared these results with those obtained by superoxide radical, hydrogen peroxide and hydroxyl radical. Isolated cardiac SR exposed to rose bengal (10 nM) irradiated at (560 nm) produced a significant inhibition of Ca2+ uptake; from 2.27 +/- 0.05 to 0.62 +/- 0.05 mumol Ca2+/mg.min (mean +/- SE) (P less than 0.01) and Ca(2+)-ATPase activity from 2.08 +/- 0.05 mumol Pi/min.mg to 0.28 +/- 0.04 mumol Pi/min.mg (mean +/- SE) (P less than 0.01). The inhibition of calcium uptake and Ca(2+)-ATPase activity by rose bengal derived activated oxygen (singlet oxygen) was dependent on the duration of exposure and intensity of light. The singlet oxygen scavengers ascorbic acid and histidine significantly protected SR Ca(2+)-ATPase against rose bengal derived activated oxygen species but superoxide dismutase and catalase did not attenuate the inhibition. SDS-polyacrylamide gel electrophoresis of SR exposed to photoactivated rose bengal up to 14 min, demonstrated complete loss of Ca(2+)-ATPase monomer band which was significantly protected by histidine. Irradiation of rose bengal also caused an 18% loss of total sulfhydryl groups of SR. On the other hand, superoxide (generated from xanthine oxidase action on xanthine) and hydroxyl radical (0.5 mM H2O2 + Fe(2+)-EDTA) as well as H2O2 (12 mM) were without any effect on the 97,000 dalton Ca(2+)-ATPase band of sarcoplasmic reticulum.(ABSTRACT TRUNCATED AT 250 WORDS)
本研究的目的是探讨单线态氧在心血管损伤中的作用。为实现这一目标，我们研究了[由孟加拉玫瑰红光激活产生的]单线态氧对心肌肌浆网钙转运和Ca(2+)-ATP酶活性的影响，并将这些结果与超氧阴离子、过氧化氢和羟自由基所获得的结果进行比较。分离的心肌肌浆网暴露于在560nm波长照射的孟加拉玫瑰红（10nM）下，Ca2+摄取受到显著抑制；从2.27±0.05降至0.62±0.05μmol Ca2+/mg·min（平均值±标准误）（P<0.01），Ca(2+)-ATP酶活性从2.08±0.05μmol Pi/min·mg降至0.28±0.04μmol Pi/min·mg（平均值±标准误）（P<0.01）。孟加拉玫瑰红衍生的活性氧（单线态氧）对钙摄取和Ca(2+)-ATP酶活性的抑制取决于暴露时间和光照强度。单线态氧清除剂抗坏血酸和组氨酸可显著保护肌浆网Ca(2+)-ATP酶免受孟加拉玫瑰红衍生的活性氧的影响，但超氧化物歧化酶和过氧化氢酶并不能减轻这种抑制作用。暴露于光激活的孟加拉玫瑰红长达14分钟的肌浆网的SDS-聚丙烯酰胺凝胶电泳显示，Ca(2+)-ATP酶单体条带完全消失，而组氨酸可对其提供显著保护。孟加拉玫瑰红的照射还导致肌浆网总巯基基团损失18%。另一方面，超氧阴离子（由黄嘌呤氧化酶作用于黄嘌呤产生）、羟自由基（0.5mM H2O2 + Fe(2+)-EDTA）以及H2O2（12mM）对肌浆网97,000道尔顿的Ca(2+)-ATP酶条带没有任何影响。（摘要截短于250字）
