复合型高水溶性二氢卟吩类光敏剂的制备及其光动力细胞治疗性能测试
Preparation of compound high water-soluble chlorin-based photosensitizer and its photodynamic cell therapy performance test
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摘要: 目的 探究一种改善二氢卟吩e6(Chlorin e6,Ce6)水溶性的简便策略,并在细胞水平对其光动力治疗性能进行测试。方法 利用生物营养素胆碱(Choline)与Ce6进行超分子复合,制备得到高水溶性光敏剂CHCe6。采用光谱法对光敏剂CHCe6的光物理性质(如紫外-可见吸收光谱、单线态氧量子产率等)进行表征。利用紫外-可见吸收光谱分析光敏剂CHCe6和Ce6在水溶液中的稳定性。通过细胞摄取实验分析鼠源性三阴乳腺癌4T1肿瘤细胞对于光敏剂CHCe6和Ce6的摄取情况。通过DHE和AO染色实验探究光敏剂CHCe6和Ce6的活性氧产生情况及其对溶酶体结构的影响。通过细胞毒性实验评价光敏剂CHCe6和Ce6对于4T1肿瘤细胞的光毒性与暗毒性。结果 通过超分子复合策略制备得到高水溶性光敏剂CHCe6。在低功率LED照射条件下,光敏剂CHCe6表现出较高的单线态氧量子产率和光稳定性。细胞摄取实验表明4T1肿瘤细胞对于光敏剂CHCe6具有较高的摄取量,并呈现时间依赖性。DHE和AO染色进一步证实,在低功率LED照射条件下光敏剂CHCe6在4T1肿瘤细胞中能高效产生活性氧,并通过破坏溶酶体结构快速转运至细胞质。细胞毒性实验表明光敏剂CHCe6表现较低的暗毒性和较高的光毒性。结论 相较于光敏剂Ce6,光敏剂CHCe6具有较高水溶性,并表现出高的光稳定性和活性氧产生能力。在低功率LED照射条件下,光敏剂CHCe6能显著抑制鼠源性三阴乳腺癌4T1细胞生长,具有较好的临床应用潜力。Abstract: Objective To explore an easy and convenient strategy to improve the solubility of chlorin e6(Ce6) in aqueous medium and evaluate its photodynamic activity at the cellular level.Methods The highly water-soluble photosensitizer CHCe6 was prepared by supramolecular combination of bionutrient choline(Choline) and photosensitizer Ce6. The photophysical properties of the photosensitizer CHCe6(including UV-Vis absorption spectrum, and singlet oxygen quantum yield) were characterized by spectrometry. The stability of photosensitizers CHCe6 and Ce6 in aqueous medium was analyzed by UV-Vis absorption spectroscopy. The cellular uptakes of photosensitizers CHCe6 and Ce6 by murine triple-negative breast cancer 4T1 tumor cells were analyzed. DHE and AO stainings were conducted to assess the production of reactive oxygen species of photosensitizers CHCe6 and Ce6 and their effects on lysosomal membrane structure. The phototoxicity and dark toxicity of photosensitizers CHCe6 and Ce6 to 4T1 tumor cells were evaluated by cytotoxicity experiments.Results The highly water-soluble photosensitizer CHCe6 was successfully prepared by the supramolecular combination strategy. CHCe6 exhibited high singlet oxygen quantum yield and photostability under low-power LED irradiation. Cellular uptake experiments showed that CHCe6 was effectively internalized by 4T1 tumor cells by a time-dependent manner. DHE and AO stainings further confirmed that CHCe6 efficiently produced reactive oxygen species in 4T1 tumor cells under low-power LED irradiation and rapidly transported to the cytoplasm through the lysosomal membrane disruption. Furthermore, cytotoxicity experiments showed that CHCe6 exhibited lower dark toxicity and higher phototoxicity.Conclusions Compared with Ce6, CHCe6 exhibits higher water solubility, with better photostability and remarkable capacity to produce reactive oxygen species. Upon low-power LED irradiation, CHCe6 can significantly inhibit the growth of murine triple-negative breast cancer 4T1 cells, demonstrating good clinical application potential.
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