(1)背景:高压氧(HBO)暴露诱导氧化应激,可能导致DNA损伤,已在人外周血淋巴细胞或非人细胞中观察到。这里,我们研究了高压条件对两种人类成骨细胞系的影响:原代人成骨细胞,HOBs,和成骨性肿瘤细胞系SAOS-2。(2)方法:将细胞在实验性高压舱中暴露于HBO(4ATA,100%氧气,37°C,和4小时)或假暴露(1ATA,空气,37°C,和4小时)。之前检查过DNA损伤,之后,以及在暴露于碱性彗星测定法并检测γH2AX53BP1共定位双链断裂(DSB)灶和凋亡后24小时。TGFβ-1,HO-1和NQO1的基因表达,参与抗氧化功能,用qRT-PCR测量。(3)结果:碱性彗星实验显示,HBO治疗4小时后,两种细胞系的DNA损伤水平均显着升高,而DSB病灶与假手术相似。γH2AX分析表明两种细胞系的细胞凋亡略有增加。直接暴露后HOB和SAOS-2中HO-1的表达增加表明在这些细胞中诱导了抗氧化反应。此外,暴露4h后,TGF-β1在HOB细胞中的表达受到负面影响。(4)结论:总之,这项研究表明,成骨细胞对高压氧的DNA损伤作用敏感,HBO诱导的DNA损伤主要由快速修复的单链DNA断裂组成。
(1) Background: Hyperbaric oxygen (HBO) exposure induces oxidative stress that may lead to DNA damage, which has been observed in human peripheral blood lymphocytes or non-human cells. Here, we investigated the impact of hyperbaric conditions on two human osteoblastic cell lines: primary human osteoblasts, HOBs, and the osteogenic tumor cell line SAOS-2. (2) Methods: Cells were exposed to HBO in an experimental hyperbaric chamber (4 ATA, 100% oxygen, 37 °C, and 4 h) or sham-exposed (1 ATA, air, 37 °C, and 4 h). DNA damage was examined before, directly after, and 24 h after exposure with an alkaline comet assay and detection of γH2AX+53BP1 colocalizing double-strand break (DSB) foci and apoptosis. The gene expression of TGFß-1, HO-1, and NQO1, involved in antioxidative functions, was measured with qRT-PCR. (3) Results: The alkaline comet assay showed significantly elevated levels of DNA damage in both cell lines after 4 h of HBO, while the DSB foci were similar to sham. γH2AX analysis indicated a slight increase in apoptosis in both cell lines. The increased expression of HO-1 in HOB and SAOS-2 directly after exposure suggested the induction of an antioxidative response in these cells. Additionally, the expression of TGF-ß1 was negatively affected in HOB cells 4 h after exposure. (4) Conclusions: in summary, this study indicates that osteoblastic cells are sensitive to the DNA-damaging effects of hyperbaric hyperoxia, with the HBO-induced DNA damage consisting largely of single-strand DNA breaks that are rapidly repaired.