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Protection Mechanism of Hydrogen Against Hyperoxia-Induced Injury in Type II Alveolar Epithelial Cells
Wu Dan1, Yao Lan2, Fang Fang1, Xu Feng1, Liu Chengjun1*
1Department of Pediatric Intensive Care Unit, Children’s Hospital of Chongqing Medical University, Chongqing 400014, China; 2Ministry of Education Key Laboratory of Child Development and Disorders; China International Science and Technology Cooperation Base of Child Development and Critical Disorders; Chongqing Key Laboratory of Pediatrics, Chongqing 400014, China
Abstract: This work was aim to investigate the protection mechanism of hydrogen against hyperoxiainduced injury in type II alveolar epithelial cells (AECII). AECII were isolated and purified from premature rats and were divided into 4 groups: air group， hyperoxia group， hyperoxia+hydrogen group， hyperoxia+hydrogen+PD98059 group. Air group and hyperoxia group were exposued to 21% oxygen and 95% oxygen， respectively. Hydrogen was added to the hyperoxia+hydrogen group before hyperoxia exposure. Hydrogen and ERK1/2 specific inhibitor PD98059 were added to hyperoxia+hydrogen+PD98059 before hyperoxia exposure， and then placed into 95% oxygen for 24 h. The ability of cell proliferation was measured by CCK-8 colorimetric assay. The cell apoptosis was analyzed by flow cytometry. The protein levels of ERK1/2， p-ERK1/2， Bax were detected by Western blot. The mRNA levels of Bax and caspase-3 were assessed by qPCR. Compared with air group， cell apoptosis rates significantly increased (P<0.01)， cells proliferation significantly decreased (P<0.01)， the mRNA levels of Bax and caspase-3 significantly increased and p-ERK1/2 significantly decreased under hyperoxia exposure (P<0.01). While with hydrogen treatment， the ability of cells proliferation markedly increased (P<0.01)， cell apoptosis rates， Bax， caspase-3 mRNA levels notably decreased (P<0.05) and p-ERK1/2 protein levels increased (P<0.05). The effect of hydrogen was abolished after treatment with inhibitor PD98059 (P<0.05). Hydrogen can inhibit the expression of apoptosis related-genes via activating the MAPK-ERK1/2 signaling pathway to improve the proliferation restriction of AECII cells induced by hyperoxia， and promote the survival of cells.
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