The idea of evidence-based medicine(EBM) has brought profound changes to clinical medicine.During clinical medicine teaching,a new method should be used that is based on the nature of evidence-based medicine.All of this is required by the trend of the times and objective reality.

OBJECTIVETo observe the effect of normobaric hyperoxia exposure on the functions of N9 microglia and explore the underlying mechanism of hyperoxia-induced immature brain injury.

METHODSN9 microglial cells were exposed to 900 ml/L O(2) for 2, 6, 12, 24 or 48 h, and the cell apoptotic rate was assessed using flow cytometry. The intracellular oxidative stress was measured using a fluorescent DCFH-DA probe, and the expression of Toll-like receptor 4 (TLR4) mRNA was detected using RT-PCR. Interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) concentrations in the supernatant of the cell cultures were tested with ELISA following the exposures. TLR4 protein expression was observed using immunofluorescence staining.

RESULTSSignificant cell apoptosis was detected after oxygen exposures for 12-24 h. Accumulation of reactive oxygen species (ROS) were detected after a 2-h exposure. After prolonged hyperoxia exposure, TLR4 expression and IL-1β and TNF-α levels significantly increased in the cells.

CONCLUSIONHyperoxia exposure activates TLR4 signaling pathway in N9 microglial cells in vitro, leading to massive production of ROS, IL-1β, and TNF-α and thus triggering cell apoptosis.

Animals ; Apoptosis ; drug effects ; Cell Hypoxia ; Cells, Cultured ; Interleukin-1beta ; metabolism ; Mice ; Mice, Inbred ICR ; Microglia ; cytology ; drug effects ; physiology ; Oxygen ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Reactive Oxygen Species ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism