目的:观察缺氧缺糖(OGD)是否损伤大鼠C6胶质瘤细胞,以及5-脂氧酶(5-LOX)/半胱氨酰白三烯(CysLT)途经是否参与OGD损伤。方法:在OGD处理并恢复不同时间后,观察C6细胞活性变化,以及5-LOX抑制剂和CysLT受体拮抗剂的影响;以免疫细胞化学法观察5-LOX蛋白的细胞内分布;以RT-PCR法检测CysLT1和CysLT2受体mRNA表达;并观察白三烯D4(LTD4)对C6细胞的作用。结果:OGD4-8h并恢复24-72h后,可诱导C6细胞损伤;5-LOX抑制剂和CysLT受体拮抗剂对OGD损伤无明显作用。OGD未诱导5-LOX的核膜移位。C6细胞高表达CysLT2受体,但CysLT1受体表达很微弱,OGD不影响它们的表达。此外,LTD4对C6细胞没有明显作用。结论:OGD可诱导C6细胞损伤,但5-LOX/CysLT途径不参与OGD诱导的损伤。
Objective: To determine whether oxygen-glucose deprivation (OGD) induces C6 cell injury,and whether 5-lipoxygenase (5-LOX)/cysteinyl leukotriene (CysLT) pathway is involved in OGD-induced injury. Methods: After OGD treatment and recovery for various durations,the viability of C6 cells was determined,and the effects of 5-LOX inhibitors and CysLT receptor antagonists were investigated. Intracellular distribution of 5-LOX protein was detected by immunocytochemistry,and the mRNA expressions of CysLT1 and CysLT2 receptors were detected by RT-PCR. The effect of leukotriene D4 (LTD4) on C6 cells was also investigated. Results: OGD for 4~8 h followed by recovery for 24~72 h significantly induced C6 cell injury. Neither 5-LOX inhibitors nor CysLT receptor antagonists inhibited OGD-induced injury. OGD did not induce 5-LOX translocation into the nuclear membrane. C6 cells highly expressed CysLT2 receptor,but the expression of CysLT1receptor was much weaker; the expression was not affected by OGD. In addition,LTD4did not affect C6 cells significantly. Conclusion: OGD can induce C6 cell injury,but 5-LOX/CysLT pathway might be not involved in OGD-induced injury.