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目的研究新型光敏剂17,4-位氨基磺酸竹红菌乙素衍生物(17-4-amino-1-butane-sulfonic acid-hypocrellin B,4SB)的光谱特性及其与人血清白蛋白(human serum albumin,HSA)的相互作用规律,为4SB临床和基础研究提供实验依据。方法使用分光光度计和荧光光谱仪测量4SB在PBS和HSA溶液中的吸收光谱和荧光光谱,并与临床使用的光敏剂血卟啉衍生物(hematoporphyrin derivative,HpD)作对比,研究HSA对4SB光谱的影响。将4SB溶于HSA溶液后持续震荡,不同的时间点测定4SB吸收光谱和荧光光谱,观察4SB与HSA作用的时间规律。观察HSA对4SB吸收和荧光光谱的影响:将浓度为0、1.25、2.5、5.0、10和20μM/ml的HSA加入5μM/ml的4SB中,测定吸收光谱和荧光光谱。HSA与4SB相互的荧光淬灭:将不同浓度的4SB加入HSA中,测定4SB的荧光光谱;将不同浓度的4SB加入HSA中,测定HSA的荧光光谱。结果 4SB在600 nm以上的波段较HpD的吸收有很大提高,在人血清溶液体系中此峰红移且增强了2.3倍。4SB与HSA的结合时间是30 min。HSA与4SB是1对1的特异性结合,且位点在色氨酸残基附近。结论 4SB在红光波段吸收好,与白蛋白结合后增强了对光的吸收,与白蛋白1∶1结合,是一种有开发前景的光敏剂。
Objective To study the spectral characteristics of 17-4-amino-1-butane-sulfonic acid-hypocrellin B (4SB), a novel photosensitizer, and its interaction with human serum albumin human serum albumin (HSA), providing experimental evidence for 4SB clinical and basic research. Methods The absorption and fluorescence spectra of 4SB in PBS and HSA were measured by spectrophotometer and fluorescence spectrophotometer. Compared with the photosensitizer hematoporphyrin derivative (HpD), the effect of HSA on 4SB influences. The 4SB was dissolved in HSA solution and then shocked continuously. The absorbance and fluorescence spectra of 4SB were measured at different time points, and the time course of the action of 4SB and HSA was observed. Observe the effect of HSA on 4SB absorption and fluorescence spectra: HSA at concentrations of 0, 1.25, 2.5, 5.0, 10 and 20 μM / ml were added to 5 μM / ml of 4SB to measure absorbance and fluorescence spectra. Fluorescence quenching between HSA and 4SB: Different concentrations of 4SB were added to HSA to measure the fluorescence spectrum of 4SB; Different concentrations of 4SB were added into HSA to measure the fluorescence spectrum of HSA. Results Compared with HpD, the absorption of 4SB in the band above 600 nm was greatly enhanced, and the peak was red-shifted by 2.3 fold in the human serum solution system. 4SB and HSA binding time is 30 min. HSA and 4SB are 1 to 1 specific binding sites near the tryptophan residue. Conclusion 4SB absorbs well in the red band and enhances the absorption of light when combined with albumin. It binds to albumin 1: 1 and is a promising photosensitizer.