背景:5-氨基乙酰丙酸光动力疗法(ALA-PDT)是毛囊皮脂腺炎性疾病的有效治疗方法,如寻常痤疮。在这项研究中,我们探讨了ALA-PDT体外抗痤疮的机制。
方法:我们用ALA(0.2mM)处理人SZ95皮脂腺细胞,并在12小时内接受不同剂量的PDT(0、5、10、20J/cm²)。我们使用AnnexinVFITC/PI凋亡试剂盒评估了治疗后的细胞活力。用DCFDA探针检测皮脂腺细胞中的ROS积累。我们通过定量PCR定量NLRP3和caspase-1mRNA,并通过ELISA测定ALA-PDT后IL-1β的释放。Western印迹有助于确定与焦亡相关的蛋白质(NLRP3,caspase-1和IL-1β)的水平。为了阐明机制,我们在给予不同浓度的NAC抗氧化剂(0,0.4,2,10mM)和半胱天冬酶抑制剂Z-VAD-FMK(0,5,10,20μM)后重新评估了这些参数.
结果:增加PDT剂量反向影响SZ95皮脂腺细胞存活,ROS和焦亡相关蛋白(NLRP3,caspase-1和IL-1β)相应增加。此外,NAC和Z-VAD-FMK以剂量响应方式调节这些分子的表达和分泌。
结论:我们的发现表明ALA-PDT对皮脂腺的潜在作用机制可能涉及ROS诱导,导致NLRP3炎性体组装,从而增强caspase-1激活和IL-1β分泌。这种级联反应可能会放大局部炎症反应,以打破寻常痤疮治疗中的慢性炎症。
BACKGROUND: 5-Aminolevulinic acid photodynamic therapy (ALA-PDT) is an effective treatment for pilosebaceous inflammatory diseases, such as acne vulgaris. In this study, we explored ALA-PDT\'s mechanisms against acne in vitro.
METHODS: We treated human SZ95 sebocytes with ALA (0.2 mM) and subjected them to varied PDT doses (0, 5, 10, 20 J/cm²) over 12 h. We assessed cell viability post-treatment using the Annexin V FITC/PI apoptosis kit. ROS accumulation in the sebocytes was detected with a DCFDA probe. We quantified NLRP3 and caspase-1 mRNA via quantitative PCR and determined IL-1β release following ALA-PDT by ELISA. Western blotting helped identify the levels of proteins associated with pyroptosis (NLRP3, caspase-1, and IL-1β). To elucidate the mechanisms, we re-evaluated these parameters after administering various concentrations of NAC antioxidants (0, 0.4, 2, 10 mM) and the caspase inhibitor Z-VAD-FMK (0, 5, 10, 20 μM).
RESULTS: Increasing PDT dose inversely affected SZ95 sebocyte survival, with a corresponding rise in ROS and pyroptosis-related proteins (NLRP3, caspase-1, and IL-1β). Furthermore, NAC and Z-VAD-FMK modulated the expression and secretion of these molecules in a dose-responsive manner.
CONCLUSIONS: Our findings suggest ALA-PDT\'s potential mechanism of action on sebaceous glands could involve ROS induction, leading to NLRP3 inflammasome assembly, thereby heightening caspase-1 activation and IL-1β secretion. This cascade may amplify the local inflammatory response to break chronic inflammation in acne vulgaris treatment.