氢气对高氧致肺泡Ⅱ型上皮细胞损伤的保护作用

摘要

Objective To investigate the protective effect of hydrogen against hyperoxia-induced oxidative stress injury in premature rat type II alveolar epithelial cells (AECs). Methods The type Ⅱ AECs isolated from premature rats were randomly divided into air (21% oxygen) control group, hyperoxia (95% oxygen) control group, air + hydrogen group, and hyperoxia+ hydrogen group. The cells with hydrogen treatment were cultured in the presence of rich hydrogen. After the corresponding exposure for 24 h, the cell morphology was observed microscopically. MTT assay was used to evaluated the cell proliferation ability, and JC-1 fluorescence probe was used to detect the mitochondrial membrane potential (ΔΨ) changes of the type Ⅱ AECs. The concentration of maleic dialdehyde (MDA) and superoxide dismutase (SOD) activity in the cell supernatant were detected using colorimetric method. Results No significant differences were found in cell growth or measurements between air control and air + hydrogen groups. Compared with air control group, the cells exposed to hyperoxia showed significantly suppressed proliferation, reduced mitochondrial membrane potential, increased MDA content, and decreased SOD activity. Intervention with hydrogen resulted in significantly increased cell proliferation and SOD activity and lowered MDA content, and restored the mitochondrial membrane potential in the cells with hyperoxia exposure (P<0.05). Conclusion Hydrogen can significantly reduce hyperoxia-induced oxidative stress injury in premature rat type II AECs, improve the cellular antioxidant capacity, stabilize the mitochondrial membrane potential, and reduce the inhibitory effect of hyperoxia on cell proliferation.%目的 探讨氢气对高浓度氧导致的早产大鼠肺泡型Ⅱ上皮细胞(AECⅡ)氧化应激性损伤的影响.方法 将分离培养的原代AECⅡ随机分为空气对照组、高氧对照组、空气+氢气组、高氧+氢气组.空气组和高氧组分别暴露在21％和95％氧气中,空气+氢气组和高氧+氢气组细胞置于富氢培养基中.各组均于24 h后光镜观察细胞形态改变,采用四甲基偶唑氮蓝(MTT)法检测各组细胞增殖能力,JC-1荧光探针法检测各组细胞线粒体膜电位(△ψ)变化,化学比色法测定细胞上清丙二醛(MDA)含量和超氧化物歧化酶(SOD)活力.结果 与空气对照组比较,空气+氢气组各项指标均无显著差异,高氧对照组细胞的增殖活性明显受到抑制,细胞的△ψ明显降低,MDA含量显著增高,SOD活性明显降低.氢气干预后,与高氧对照组相比,细胞的增殖活性和SOD活性均有所升高,△ψ有所恢复,MDA含量显著下降(P＜0.05).结论 氢气可显著减轻高氧导致的早产鼠AECⅡ的氧化损伤,提高细胞抗氧化能力,稳定细胞△ψ,减弱高氧对细胞的增殖抑制效应.