齐留通减轻小鼠小胶质细胞介导的鱼藤酮对PC12细胞的毒性作用

Zileuton, a 5-lipoxygenase inhibitor, attenuates mouse microglial cell-mediated rotenone toxicity in PC12 cells

目的：观察5-脂氧酶选择性抑制剂齐留通对小胶质细胞介导的鱼藤酮神经毒性的影响.方法：以鱼藤酮预处理的小鼠小胶质BV2细胞条件培养液培养PC12细胞,通过MTT法和乳酸脱氢酶（LDH）释放分析PC12细胞损伤和活性变化,Hoechst/碘化丙啶荧光双染检测PC12细胞死亡,并评估齐留通对小胶质细胞介导细胞毒性的作用.结果：1、3、10 nmol/L鱼藤酮对PC12细胞无直接毒性,但是其预处理的BV2细胞条件培养液间接诱导PC12细胞形态改变、活性下降,LDH释放和细胞死亡明显增加;0.01、1μmol/L齐留通能有效减轻小胶质细胞介导的鱼藤酮细胞毒性作用.结论：5-脂氧酶抑制剂齐留通能减轻小胶质细胞介导的鱼藤酮神经毒性,提示5-脂氧酶通路在小胶质细胞炎症诱导的神经细胞死亡中有重要作用.

Objective：To examine the effect of a selective inhibitor of 5-lipoxygenase （5-LOX） zileuton on microglia-mediated rotenone neurotoxicity.Methods：The supernatant from different concentrations of rotenone-stimulated mouse microglia BV2 cells was used as the conditioned media （CM） for PC12 cells.The viability of PC12 cells was determined by MTT assay and lactate dehydrogenase （LDH） release.Cell death was observed by LDH release and double fluorescence staining with Hoechst/propidiumiodide （PI）.The effect of zileuton on microglia-mediated rotenone toxicity was evaluated by the above methods.Results：Rotenone at 1-10 nmoL/L was nontoxic to PC12 cells directly.However,the CM from BV2 cells that were treated with rotenone （1-10 nmol/L） resulted in toxicity of PC12 cells.The BV2 CM which stimulated with rotenone （1-10 nmol/L） induced morphological changes,reduced cell viability,and increased LDH release and cell necrosis in PC12 cells.Pretreatment of BV2 cells with the 5-LOX inhibitor zileuton （0.01-1 μmol/L） protected PC12 cells from the microglia-mediated rotenone toxicity.Conclusion：The 5-LOX inhibitor zileuton effectively attenuates microglia-mediated rotenone toxicity in PC12 cells.These results suggest that 5-LOX pathway may be involved in neuronal death induced by microglial inflammation.