Sofic E, Rimpapa Z, Kundurovic Z, Sapcanin A, Tahirovic I, Rustembegovic A, Cao G
Neuroscience Laboratory, Jean Mayer USDA Human Nutrition Research Center on Aging, Tufts University, Boston, MA, USA.
J Neural Transm (Vienna). 2005 Mar;112(3):349-58. doi: 10.1007/s00702-004-0270-4. Epub 2005 Jan 24.
The aim of this study was to elucidate the antioxidant behaviour of melatonin (M) and determine its activity-structure relationship. M or 5-metoxy-N acetyltriptamine is a neurohormone secreted by the pineal gland, which plays a proven role in maintaining sleep-wake rhythms. The antioxidant capacity of M was analysed using the oxygen radical absorbance capacity (ORAC) assay. Furthermore, spectral measurements for aerobic photolytic reaction of neutral red (NR) and degree of inhibition of photolysis with M, glutathione (GSH), ascorbic acid (AA) and vitamin E analogue Trolox were studied at room temperature 25 degrees C, using visible (VIS) and ultra-violet (UV) radiations. In the ORAC assay 2,2-azobis (2-amidino-propane)dihydrochloride (AAPH) a peroxyl radical generator, ROO degrees ; H2O2-Cu2+, mainly a hydroxyl radical generator, degrees OH; and Cu2+ a transition metal were used. Although some studies indicated that M is a powerful antioxidant, no one has compared its antioxidant capacities with GSH, E-vitamin and AA, using three free radical (FR) generators in an assay which utilizes an area-under curve technique and thus combines both inhibition time and inhibition degree of FR action by an antioxidant into a single quantity. In the current study, we used ORAC assay with three FR generators. The assay is based on propensity of the fluorescence emitted by the protein beta-phycoerythrin (beta-PE) from porphyridium cruentum to be quenched when exposed to FR action. M in our experiments acted as a universal antioxidant against ROO degrees and degrees OH radicals. Also, M served as an antioxidant in the presence of Cu2+. M, which is a lipid-soluble compound, was a twice more powerful antioxidant than vitamin E, and four times than AA or GSH. Furthermore, M inhibited aerobic photolysis of NR photoinduced with VIS and UV rays faster and more effectively, than AA, GSH or vitamin E. AA with NR, under aerobic conditions during irradiation with VIS and UV acted as a pro-oxidant. M may be the premier molecule to protect the cells from oxidative stress.
本研究的目的是阐明褪黑素(M)的抗氧化行为并确定其活性与结构的关系。M即5-甲氧基-N-乙酰色胺,是一种由松果体分泌的神经激素,在维持昼夜节律方面发挥着已被证实的作用。采用氧自由基吸收能力(ORAC)测定法分析了M的抗氧化能力。此外,在25℃室温下,使用可见光(VIS)和紫外线(UV)辐射,研究了中性红(NR)的需氧光解反应的光谱测量以及M、谷胱甘肽(GSH)、抗坏血酸(AA)和维生素E类似物Trolox对光解的抑制程度。在ORAC测定中,使用了过氧自由基发生器2,2-偶氮二(2-脒基丙烷)二盐酸盐(AAPH),即ROO˙;主要作为羟基自由基发生器的H2O2-Cu2+,即˙OH;以及过渡金属Cu2+。尽管一些研究表明M是一种强大的抗氧化剂,但在利用曲线下面积技术的测定中,没有人使用三种自由基(FR)发生器将其抗氧化能力与GSH、维生素E和AA进行比较,该技术将抗氧化剂对FR作用的抑制时间和抑制程度结合为一个单一量。在本研究中,我们使用了带有三种FR发生器的ORAC测定法。该测定基于来自紫球藻的蛋白质β-藻红蛋白(β-PE)发出的荧光在暴露于FR作用时被淬灭的倾向。在我们的实验中,M对ROO˙和˙OH自由基起到了通用抗氧化剂的作用。此外,在Cu2+存在的情况下,M也作为抗氧化剂发挥作用。M是一种脂溶性化合物,其抗氧化能力是维生素E的两倍,是AA或GSH的四倍。此外,M比AA、GSH或维生素E更快、更有效地抑制了由VIS和UV光诱导的NR的需氧光解。在VIS和UV辐射的需氧条件下,AA与NR一起作为促氧化剂。M可能是保护细胞免受氧化应激的首要分子。