【摘要】 目的:探讨血卟啉单甲醚光动力治疗(hematoporphyrin monomethyl ether photodynamic therapy,HMME-PDT)对人真皮微血管内皮细胞氧化应激损伤的具体作用及可能涉及的机制,为HMME-PDT治疗葡萄酒色斑的临床应用提供理论依据。方法:采用HDMECs 10～15代用于本次实验。通过CCK-8检测细胞活性变化;流式细胞术检测各组细胞凋亡比例及细胞内活性氧簇(reactive oxygen species,ROS)水平;分光光度计法检测各组细胞培养上清液乳酸脱氢酶水平及丙二醛水平;采用方差分析比较各组差异。Western blot检测各组c-Jun氨基末端激酶(c-Jun N-terminal Kinase,JNK)、P38、细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)总蛋白及磷酸化蛋白表达。结果:HMME-PDT能明显降低人真皮微血管内皮细胞活性,诱导细胞凋亡(P<0.05)。HMME-PDT能诱导人真皮微血管内皮细胞产生大量ROS(P<0.05)。HMME-PDT可以诱导P38、ERK磷酸化。结论:HMME-PDT能诱导人真皮微血管内皮细胞的氧化损伤,其机制可能与激活MAPKs通路有关。更多还原

【Abstract】 Objective: To investigate the effects and mechanism of hematoporphyrin monomethyl ether(HMME) photodynamic therapy(PDT) induced oxidative stress in human dermal microvascular endothelial cells(HDMECs) to provide a theoretical basis for the therapeutic opportunities in port-wine stains by HMME-PDT. Methods: The 10 th-15 th generation of HDMECs were used for all experiments. Changes in cell viability were detected by CCK-8; Intracellular and apoptosis reactive oxygen species(ROS) levels were investigated by flow cytometry; lactate dehydrogenase and malonaldehyde contents were evaluated by spectrophotometry; and analysis of variance was used to compare the differences in groups. Total protein and phosphorylated protein of JNK, P38, ERK were detected by Western blot. Results: HMME-PDT significantly reduced the activity of HDMECs(P<0.05), and induced apoptosis(P<0.05), intracellular ROS(P<0.05) and P38, ERK phosphorylation. Conclusion: HMME-PDT can significantly induce oxidative stress in HDMECs, which may be associated with the activation of MAPKs.更多还原