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光动力疗法后芦荟大黄素负载脂质体对黑色素瘤细胞系的体外光敏疗效研究

The examination of in vitro photosensitizing efficacy of aloe-emodin loaded liposome following photodynamic therapy on melanoma cell lines.

作者信息

Nowak-Perlak Martyna, Olszowy Marta, Kupczyk Piotr, Wrześniok Dorota, Zaremba-Czogalla Magdalena, Dębicka Roksana, Makarec Alicja, Gubernator Jerzy, Ziółkowski Piotr, Woźniak Marta

机构信息

Department of Clinical and Experimental Pathology, Division of General and Experimental Pathology, Wroclaw Medical University, Wroclaw, 50-368, Poland.

Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences in Sosnowiec, Medical University of Silesia in Katowice, 4 Jagiellońska, Sosnowiec, 41-200, Poland.

出版信息

Sci Rep. 2025 Aug 6;15(1):28833. doi: 10.1038/s41598-025-14777-4.

Abstract

This study focused on investigating the enhanced photoactive properties of liposomal aloe-emodin for photodynamic treatment (PDT). Aloe-emodin (A-E) was encapsulated in liposomes by using the copper ion gradient method. This study aimed to highlight the function of naturally occurring photoactive plant-based derivatives in liposomal formulations as simple alternative photosensitizers (PS) for melanoma photodynamic therapy (PDT). Furthermore, we aimed to show that phototoxic chemicals loaded into liposomes are less cytotoxic than non-encapsulated molecules, which we used in our previous study. To assess the effectiveness of liposome aloe-emodin as a photosensitizer in PDT on melanoma cell lines, we performed cellular uptake by flow cytometry, MTT assay to measure the cytotoxicity of a natural compound, wound healing assay, ROS analysis by confocal microscopy, ROS analysis by flow cytometry, immunocytochemistry staining to measure the level of apoptotic proteins, and Real-Time PCR gene expression analysis. The melanoma cells showed increased phototoxicity after therapy compared to the normal cells in the MTT assay. Moreover, liposome aloe-emodin-based photodynamic therapy resulted in an increased ratio of apoptotic cells, increased ROS levels, and increased or decreased gene expression. This study also demonstrated that aloe-emodin encapsulated in liposomes significantly decreased melanoma cell motility following PDT treatment. In conclusion, our study provides evidence that aloe-emodin-mediated PDT exhibits potential for clinical development as a novel, safe, and effective photosensitizer for melanoma treatment.

摘要

本研究聚焦于探究脂质体芦荟大黄素用于光动力治疗(PDT)时增强的光活性特性。采用铜离子梯度法将芦荟大黄素(A-E)包封于脂质体中。本研究旨在突出脂质体制剂中天然存在的具有光活性的植物源性衍生物作为黑色素瘤光动力疗法(PDT)的简单替代光敏剂(PS)的功能。此外,我们旨在表明,负载于脂质体中的光毒性化学物质的细胞毒性低于未包封的分子,这一点我们在之前的研究中已有所应用。为评估脂质体芦荟大黄素作为PDT中黑色素瘤细胞系光敏剂的有效性,我们通过流式细胞术进行细胞摄取实验、采用MTT法测定天然化合物的细胞毒性、进行伤口愈合实验、通过共聚焦显微镜进行活性氧(ROS)分析、通过流式细胞术进行ROS分析、进行免疫细胞化学染色以测定凋亡蛋白水平以及进行实时聚合酶链反应(Real-Time PCR)基因表达分析。在MTT实验中,与正常细胞相比,黑色素瘤细胞在治疗后表现出增强的光毒性。此外,基于脂质体芦荟大黄素的光动力疗法导致凋亡细胞比例增加、ROS水平升高以及基因表达增加或减少。本研究还表明,PDT处理后,脂质体包封的芦荟大黄素显著降低了黑色素瘤细胞的运动能力。总之,我们的研究提供了证据,表明芦荟大黄素介导的PDT作为一种用于黑色素瘤治疗的新型、安全且有效的光敏剂具有临床开发潜力。

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