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将两类水难溶性金属卟啉(MP),meso-四苯基卟啉金属配合物(MTPP,M=ZnII,CoII)和meso-四(p-羟基苯基)卟啉金属配合物(MTpHPP,M=ZnII,CoII)分别与牛血清白蛋白(BSA)结合,制得水溶性金属卟啉白蛋白结合体(MP@BSA).采用UV-vis光谱、聚丙烯酰胺凝胶电泳(PAGE)、圆二色谱进行了表征与分析,发现金属卟啉的水溶性和稳定性得到大幅度提高.采用NBT光还原法测定了MP@BSA结合体清除超氧阴离子自由基(O2·-)的能力,发现结合体MP@BSA具有清除O2·-的性能,与小分子金属卟啉相比,结合体的抗O2·-性能提高了一个数量级.MP@BSA抗氧化活性分别是VC和BSA的600倍和40倍.其中,羟基取代金属卟啉结合体表现出更强的清除O2·-的活性,CoTpHPP@BSA的EC50为1.5μmol/L,对天然Cu,Zn-SOD的模拟度为2.73%.最后提出了生物高分子结合体清除O2·-的可能机理.
Two kinds of metal complexes of water-insoluble metalloporphyrin (MP), meso-tetraphenylporphyrin (MTPP, M = ZnII, CoII) and meso-tetrakis (p- hydroxyphenyl) porphyrin , M = ZnII, CoII) were respectively combined with bovine serum albumin (BSA) to prepare a water-soluble metalloporphyrin albumin complex (MP @ BSA) .Using UV-vis spectroscopy, polyacrylamide gel electrophoresis , Circular dichroism spectroscopy was used to characterize and analyze the water-soluble metalloporphyrins and their stability was greatly improved. The ability of MP @ BSA conjugates to scavenge superoxide anion radicals (O2 · -) was measured by NBT photoreduction method , And found that the combination MP @ BSA has the ability to scavenge O2 ·, and the anti-O2 · -adjunction of the conjugate is improved by one order of magnitude compared with the small molecule metalloporphyrin.MP @ BSA antioxidant activity is 600 Times and 40 times, respectively.The hydroxyl-substituted metalloporphyrins showed stronger O2 · - removal activity, the EC50 of CoTpHPP @ BSA was 1.5μmol / L, the mimics of natural Cu and Zn-SOD were 2.73% Finally, the possible mechanism of biological macromolecule combination eliminating O2 · - is proposed.