Effects Of Hypoxia On The Expression Of IL-1β、IL-6and TNF-α For Cerebral Cortex Cells In SD Rats

ObjectiveTo investigate the effect of hypoxia on the expression of IL-1β、TNF-α andIL-6,primary cultured cerebral cortex cells were treated for different time under dif-ferent oxygen concentration, then IL-1β、TNF-α and IL-6protein and IL-1β、TNF-α�'�IL-6mRNAwere tested.MethodsCultured rat cerebral cortex cells in vitro were seeded into48-well cell cultureplate at5.5-7.0×105cells/ml cell density. Primary cultured cells were divided intotwo groups: hypoxic and control. The cells subjected to hypoxia were further dividedinto four groups: Cells were subjected to1%oxygen concentration for3h,and markedas H3group; Cells were subjected to1%oxygen concentration for6h,and marked asH6group;Similarly, cells were subjected to4%oxygen concentration for3h and alsomarked as H3group; cells were subjected to4%oxygen concentration for6h and alsomarked as H6group.Each hypoxic treatment group had a corresponding normoxiacontrol, and was marked as C3or C6. Cerebral cortex cells which were cultured for5days were subjected to hypoxia. After exposure to hypoxia, culture medium samplesand cells were collected. The production of IL-1β, IL-6and TNF-α mRNA were ana-lyzed by RT-QPCR, and Enzyme-linked immunosorbent assay(ELISA) was used todetermine the levels of IL-1β, IL-6and TNF-α. Each experiment was repeated threetimes independently.Results1.The effect of hypoxia(1%oxygen concentration) on the secretion of IL-1β、IL-6and TNF-α: Compared with the corresponding normoxia controls, the levels ofIL-6of H3or H6decreased significantly (P<0.05),but the levels of IL-1β�'�TNF-αhad no significant changes(P>0.05).2. The effect of hypoxia(1%oxygen concentration) on the expression of IL-1β、IL-6and TNF-α mRNA: Compared with the corresponding normoxia controls, thelevels of IL-1β and TNF-α mRNA of H3or H6had no significant changes(P>0.05);Compared with C3, the level of IL-6mRNA of H3had a trend to rise, compared withC6, the level of IL-6mRNA of H6had a significant increase(P<0.05).3. The effect of hypoxia(4%oxygen concentration) on the secretion of IL-1β、IL-6and TNF-α: Compared with C3, the levels of IL-1β and IL-6of H3had a trend to decrease, and compared with C6, the levels of IL-1β and IL-6of H6significantly de-creased;(P<0.05). Compared with the corresponding normoxia controls, the levels ofTNF-α mRNA of H3or H6had no significant changes(P>0.05).4.The effect of hypoxia(1%oxygen concentration) on the expression of IL-1β、IL-6and TNF-α mRNA: Compared with the corresponding normoxia controls C3andC6, the levels of IL-1β and TNF-α mRNA of H3or H6had no significantchanges(P>0.05); Compared with C3, the level of IL-6mRNA of H3had a trend torise, and compared with C6, the level of IL-6mRNA of H6had a significant in-crease(P<0.05).Conclusion1. Oxygen at1%has no influence on IL-1β or TNF-α secretion in cerebral cortexcells of rats, and it can inhibit the production of IL-6.2. Oxygen at1%has no influence on IL-1β or TNF-α mRNA production in cere-bral cortex cells of rats, and the stimulation of oxygen at1%on the production of IL-6mRNA has a time-dependent.3. Oxygen at4%has no effect on TNF-α production, and the inhibitory effects onIL-1β and IL-6have a time-dependent.4. Oxygen at4%has no effect on IL-1β or TNF-α mRNA production, and thestimulation of oxygen at4%on the production of IL-6mRNA has a time-dependent.