研究目的
To evaluate the effects of direct exposure to non-equilibrium atmospheric pressure plasma (NEAPP) on biomolecules, including lipids, proteins, and nucleic acids, and to standardize these oxidative modifications for potential biomedical applications, particularly in cancer treatment.
研究成果
Direct exposure to NEAPP induces simultaneous oxidative and UV-specific damage in biomolecules, including lipids, proteins, and DNA, in a dose-dependent manner. Hydroxyl radicals are a major effector, and the damage can be controlled site-specifically. This suggests potential applications in cancer treatment, such as eradicating surface cancer cells, but requires further study for clinical translation.
研究不足
The oxidative stress induced by NEAPP is relatively mild, as GSH levels did not decrease significantly. The study focused on in vitro and ex vivo models, which may not fully replicate in vivo conditions. The depth of effect is limited to a few millimeters in tissue, and further research is needed to understand DNA repair mechanisms and the precise fractions of reactive species generated.
1：Experimental Design and Method Selection:
The study employed a NEAPP device with high electron density to generate reactive species. Established techniques in free radical biology, such as ESR spin-trapping, spectrophotometry, ELISA, and immunohistochemistry, were used to detect oxidative and UV-induced modifications.
2：Sample Selection and Data Sources:
Samples included linoleic acid, α-linolenic acid, phosphatidylcholine, liposomes from vitamin E-stripped corn oil, rat liver homogenates, plasmid DNA, and rat genomic DNA. Animal experiments used F1 hybrid rats bred from Fischer-344 and Brown-Norway strains.
3：List of Experimental Equipment and Materials:
Key equipment included a NEAPP device, ESR spectrometer (JEOL FR-30), spectrophotometer (GeneQuant 1300), fluorometric assay system (PowerScan4), and imaging system (BZ-9000). Chemicals and kits were sourced from various suppliers, e.g., Oxford Biomedical Research, Wako, Sigma-Aldrich.
4：0). Chemicals and kits were sourced from various suppliers, e.g., Oxford Biomedical Research, Wako, Sigma-Aldrich.
Experimental Procedures and Operational Workflow:
4. Experimental Procedures and Operational Workflow: Samples were exposed to NEAPP in 96-well plates for specified durations (e.g., 30 s to 5 min). Post-exposure, analyses were conducted: ESR for hydroxyl radicals, absorbance at 234 nm for conjugated dienes, TBARS assay for lipid peroxidation, GSH measurement, immunohistochemistry for protein modifications, gel electrophoresis for DNA strand breaks, and ELISA for 8-OHdG and CPDs.
5：Data Analysis Methods:
Statistical analyses used one-way ANOVA and unpaired t-tests with GraphPad Prism software. Data are expressed as mean ± SEM, with significance at p<0.05.
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