Validation of lucigenin as a chemiluminescent probe to monitor vascular superoxide as well as basal vascular nitric oxide production
Validation of lucigenin as a chemiluminescent probe to monitor vascular superoxide as well as basal vascular nitric oxide production
复制标题
DOI：
10.1006/bbrc.1998.9942
复制
发表时间：
1999-01-19
影响因子：
3.1
通讯作者：
Münzel, T
中科院分区：
文献类型：
--
作者：
Skatchkov, MP;Sperling, D;Münzel, T
光泽精已被广泛用作血管超氧阴离子（O-2（.-））监测的荧光底物阵光泽精检测O-2（.-）的有效性由于O-2（.-）是由光泽精本身产生的。已表明光泽精的浓度是影响该测定法有效性的关键参数。在目前的研究中，我们评估了光泽精（5 μ M）作为定量O-2（.-）在维管组织中产生。光泽精诱导的内皮功能的影响进行了评估，通过等距张力记录的离体主动脉环悬挂在器官浴。光泽精对O-2（.-）的影响使用自旋捕获和电子自旋共振光谱研究了产物。光泽精在250 μ M而不是在5 μ M时，引起内皮依赖性舒张乙酰胆碱的显着衰减，这是通过用超氧化物歧化酶预处理来防止的。自旋捕获研究表明，光泽精在250 μ M时增加血管O-2（.-）而5 μ M光泽精不能刺激O-2（.-）生产N-G-单甲基-L-精氨酸对NO合酶的抑制以及内皮的去除几乎使光泽精衍生的化学发光（LDCL）加倍，表明内皮衍生的NO的基础产生抑制基线化学发光信号。因此，光泽精在5 μ M的浓度似乎是一个敏感和有效的探针评估O-2（.-）在血管组织中。它也可以用作间接探针来估计基础血管NO释放。(C)北京：科学出版社.
Lucigenin has been widely used as a chemiluminescent substrate to monitor vascular superoxide (O-2(.-)) formation. The validity of lucigenin for detection of O-2(.-) has been questioned because O-2(.-) is generated by lucigenin itself. It has been shown that the concentration of lucigenin is a critical parameter affecting the validity of this assay. In the present studies we evaluated a reduced concentration of lucigenin (5 mu M) as a tool to quantify O-2(.-) production in vascular tissue. Lucigenin-induced effects on endothelial function were assessed by isometric tension recording of isolated aortic rings suspended in organ baths. The effects of lucigenin on O-2(.-) production were studied using spin trapping and electron spin resonance spectroscopy. Lucigenin at 250 mu M but not at 5 mu M caused a significant attenuation of endothelium-dependent relaxations to acetylcholine, which was prevented by pretreatment with superoxide dismutase, Spin-trapping studies revealed that lucigenin at 250 mu M increased vascular O-2(.-) production several fold while 5 mu M lucigenin did not stimulate O-2(.-) production. Inhibition of NO synthase by N-G-momomethyl-L-arginine as well as the removal of the endothelium almost doubled lucigenin-derived chemiluminescence (LDCL), indicating that basal production of endothelium-derived NO depresses the baseline chemiluminescence signal. Thus, lucigenin at a concentration of 5 mu M seems to be a sensitive and valid probe for assessing O-2(.-) in vascular tissue. It can also be used as an indirect probe to estimate basal vascular NO release. (C) 1999 Academic Press.