Aims: Irradiation-induced damage to pulmonary endothelial cells is thought to be an important mediator of the pathogenesis of radiation pneumonopathy. Tetrahydropalmatine (THP) has been shown to have a protective effect against oxidative stress. This study was designed to investigate the potential radioprotective effect of THP against irradiation-induced endothelial cellular damage and to elucidate the underlying mechanisms.
Main Methods: Human EA.hy926 cells were treated with THP and irradiation. Cell viability was measured using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. For the detection of apoptosis, morphological observation, flow cytometry and a caspase-3 activity assay were employed. The expression of cytochrome-c and Bax/Bcl-2 protein were detected by western blot analysis. Generation of reactive oxygen species (ROS) was measured by flow cytometry. Malondialdehyde (MDA), lactate dehydrogenase (LDH), glutathione (GSH) and superoxide dismutase (SOD) were measured to assess cellular oxidative stress induced injury.
Key Findings: Preincubation of EA.hy926 cells with THP before gamma-radiation resulted in significant inhibition of apoptosis and enhancement of cell viability, as revealed by morphological observation, flow cytometry and MTT assay. THP significantly reduced intracellular ROS formation, levels of intracellular MDA and LDH, and enhanced the production of intracellular antioxidants (GSH and SOD) in EA.hy926 cells. Meanwhile, THP also inhibited the decrease of intracellular mitochondrial membrane potential (psim), caspase-3 activation, cytochrome-c release and reduced Bax/Bcl-2 ratio in THP pretreated, irradiated cells.
Significance: Our findings demonstrated THP could effectively protect endothelial cells against gamma-irradiation injury, which could potentially be applied to the prevention of endothelial cell dysfunctions associated with ionizing irradiation-induced lung injury.
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