Kansai Institute for Photon Science, National Institutes for Quantum Science and Technology (QST), Kizugawa 619-0215, Japan.
Kansai Institute for Photon Science, National Institutes for Quantum Science and Technology (QST), Kizugawa 619-0215, Japan.
J Photochem Photobiol B. 2024 Sep;258:112994. doi: 10.1016/j.jphotobiol.2024.112994. Epub 2024 Jul 23.
We investigated the deoxyribonucleic acid (DNA) damage induced by laser filamentation, which was generated by focusing femtosecond near-infrared Ti:Sapphire laser light in water at several repetition rates ranging from 1000 Hz to 10 Hz. Using plasmid DNA (pUC19), the single-strand break, double-strand break, nucleobase lesions, and the fragmented DNA were analyzed and quantified by agarose gel electrophoresis. Additionally, the HO concentration after irradiation was determined. We observed that (1) the DNA damage per laser shot and (2) the enzyme-sensitive base lesions per total DNA damage decreased as the laser repetition rate increased. Furthermore, (3) extraordinarily short DNA fragments were likely to be produced, compared with those produced using X-rays, and (4) most OH radicals could be eliminated by recombination to generate HO, preventing them from damaging the DNA. The Monte-Carlo simulation of the strand break formation implies that the observed dependency of strand break efficiency on the laser repetition rate is mainly due to diffusion of DNA molecules. These findings quantitatively and qualitatively revealed that an intense laser pulse induces a specific DNA damage profile that is not induced by X-rays, a sparsely ionizing radiation source.
我们研究了由飞秒近红外钛宝石激光在水中聚焦产生的光丝化诱导的脱氧核糖核酸(DNA)损伤,激光重复率范围从 1000 Hz 到 10 Hz。我们使用质粒 DNA(pUC19)通过琼脂糖凝胶电泳分析和定量了单链断裂、双链断裂、核碱基损伤和碎片化的 DNA。此外,还测定了辐照后 HO 的浓度。我们观察到:(1)随着激光重复率的增加,每个激光脉冲的 DNA 损伤和(2)总 DNA 损伤中酶敏感碱基损伤的比例降低。此外,(3)与 X 射线相比,可能产生极短的 DNA 片段,(4)大多数 OH 自由基可以通过重组生成 HO 而被消除,从而防止它们损伤 DNA。链断裂形成的蒙特卡罗模拟表明,观察到的链断裂效率对激光重复率的依赖性主要是由于 DNA 分子的扩散。这些发现从定量和定性两方面揭示了强激光脉冲诱导的特定 DNA 损伤谱,而这种损伤谱不会由 X 射线这种稀疏电离辐射源引起。