研究目的
Investigating the effect of chlorin e6-mediated photodynamic therapy (Ce6-PDT) with halogen light on lipogenesis in human sebocytes, specifically its potential as a sebostatic treatment for acne vulgaris.
研究成果
Ce6-PDT with halogen light significantly reduces lipid synthesis in human sebocytes by modulating Akt/mTOR and AMPK/PPARγ/C/EBPα signaling pathways. This demonstrates its potential as a sebostatic therapy for acne vulgaris, offering a safer alternative to existing treatments with fewer side effects.
研究不足
The study is conducted in vitro on human sebocyte cell lines, which may not fully replicate in vivo conditions of acne vulgaris. The use of halogen light and specific Ce6 concentrations might have limitations in penetration and efficacy in clinical settings. Potential side effects or long-term impacts of Ce6-PDT were not assessed.
1：Experimental Design and Method Selection:
The study used Ce6-PDT with halogen light to treat human sebocytes pre-treated with linoleic acid (LA) to induce lipogenesis. Methods included oil red O staining, triglyceride and cholesterol assays, and western blot analysis to assess lipid content and molecular signaling pathways.
2：Sample Selection and Data Sources:
Human sebocytes were obtained from Celprogen Inc. and cultured in sebocyte complete medium. Cells were pre-treated with LA to stimulate lipogenesis before Ce6-PDT application.
3：List of Experimental Equipment and Materials:
Equipment included a 6-well cell culture plate (Falcon, Corning), halogen light source (5,000 lux), CCD camera (IMT cam CCD Pro2, IMT i-Solution Inc.), ELISA reader (Sunrise, Tecan), microplate reader (Sunrise, Tecan), and western blot apparatus (Supernova ECL system, Cyanagen s.r.l.; Sensi-Q 2000, Lugen Sci Co. Ltd.). Materials included Ce6, oil red O stain, triglyceride and cholesterol assay kits (Abcam), primary and secondary antibodies, RIPA buffer, SDS-PAGE gels, and PVDF membranes.
4：Experimental Procedures and Operational Workflow:
Sebocytes were seeded, pre-treated with LA for 24h, treated with Ce6 for 30min, exposed to halogen light for 30min, incubated in the dark, and then assessed for lipid content and protein expression after additional incubation. Staining and assays were performed according to standard protocols, with optical density measurements and western blot analysis.
5：Data Analysis Methods:
Data were analyzed using SPSS version 20 with one-way ANOVA followed by Tukey's test. Band intensities from western blots were quantified using ImageJ software.
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6-well cell culture plate
Falcon
Corning
Used for culturing sebocytes in the experiment
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CCD camera
IMT cam CCD Pro2
IMT i-Solution Inc.
Used for photographing stained cells in oil red O staining
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ELISA reader
Sunrise
Tecan
Used for measuring optical density in lipid content determination and assays
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Microplate reader
Sunrise
Tecan
Used for measuring optical densities in triglyceride and cholesterol assays
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Western blotting detection system
Supernova ECL
Cyanagen s.r.l.
Used for visualizing proteins in western blot analysis
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Imaging system
Sensi-Q 2000
Lugen Sci Co. Ltd.
Used for taking images in western blot analysis
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Triglyceride Quantification Assay Kit
Abcam
Used for measuring triglyceride levels in sebocytes
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Cholesterol Assay Kit
Abcam
Used for measuring cholesterol levels in sebocytes
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