Xue Jingwen, Gruber Florian, Tschachler Erwin, Zhao Yi
Department of Dermatology, Beijing Tsinghua Changgung Hospital, School of Clinical Medicine, Tsinghua University, Beijing 102218, China.
Department of Dermatology, Medical University of Vienna, Vienna, Austria.
Photodiagnosis Photodyn Ther. 2021 Mar;33:102137. doi: 10.1016/j.pdpdt.2020.102137. Epub 2020 Dec 8.
Photodynamic therapy (PDT) provides a treatment for port-wine stain (PWS) using hemoporfin (hematoporphyrin monomethyl ether, HMME), a novel photosensitizer, reporting better efficacy and lower recurrence rate. This study investigated the effects of HMME-PDT on human umbilical vein endothelial cells (HUVECs) as well as underlying mechanisms.
Cell proliferation ability was measured by CCK8 assay and cell apoptosis was determined by TUNEL assay and Western blot analysis. Confocal fluorescence microscopy monitoring RFP-GFP-LC3 transfected HUVECs and Western blot analysis were used to evaluate autophagy. 3-Methyladenine (3-MA), Z-VAD-FMK, N-acetylcysteine (NAC) were used for inhibitor studies.
HMME-PDT decreased cell proliferation ability in an HMME concentration and light dose-dependent manner. Oxidative stress played an important role in HMME-PDT induced cell apoptosis and autophagy in HUVECs. Pretreatment with Z-VAD-FMK, the inhibitor of apoptosis, enhanced HMME-PDT induced autophagy. 3-MA, the suppressor of autophagy, significantly increased HMME-PDT induced apoptosis rates.
Our study demonstrated that HMME-PDT induced both apoptosis and autophagy in HUVECs via oxidative stress. Our data suggested that HMME-PDT- induced autophagy was able to prevent apoptotic cell death of HUVECs and rendered them more resistant to HMME-PDT induced toxicity.
光动力疗法（PDT）使用新型光敏剂血卟啉单甲醚（HMME）治疗鲜红斑痣（PWS），疗效更佳且复发率更低。本研究调查了HMME-PDT对人脐静脉内皮细胞（HUVECs）的影响及其潜在机制。
采用CCK8法检测细胞增殖能力，通过TUNEL法和蛋白质免疫印迹分析确定细胞凋亡情况。利用共聚焦荧光显微镜监测红色荧光蛋白-绿色荧光蛋白-微管相关蛋白1轻链3（RFP-GFP-LC3）转染的HUVECs并进行蛋白质免疫印迹分析，以评估自噬。使用3-甲基腺嘌呤（3-MA）、Z-VAD-FMK、N-乙酰半胱氨酸（NAC）进行抑制剂研究。
HMME-PDT以HMME浓度和光剂量依赖性方式降低细胞增殖能力。氧化应激在HMME-PDT诱导的HUVECs细胞凋亡和自噬中起重要作用。凋亡抑制剂Z-VAD-FMK预处理增强了HMME-PDT诱导的自噬。自噬抑制剂3-MA显著提高了HMME-PDT诱导的凋亡率。
我们的研究表明，HMME-PDT通过氧化应激诱导HUVECs细胞凋亡和自噬。我们的数据表明，HMME-PDT诱导的自噬能够防止HUVECs细胞凋亡死亡，并使其对HMME-PDT诱导的毒性更具抗性。
