褐藻多糖硫酸酯在小鼠N2a细胞缺血再灌注损伤中的作用

Effect of Fucoidan on Ischemia-reperfusion Injury of N2a Cells in Mice

目的观察褐藻多糖硫酸酯(FPS)对氧糖剥夺/复糖复氧(OGD/R)诱导小鼠N2a细胞缺血再灌注损伤中的作用并探讨其可能的分子机制。方法在细胞水平上,建立小鼠N2a细胞的OGD 2 h后复灌24 h的缺血再灌注损伤模型。实验分为对照组、OGD/R模型组和FPS处理组(1、2、5、10μg/ml)。3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)比色法和乳酸脱氢酶(LDH)释放率试剂盒分别检测各组细胞存活率和死亡率;Western blot检测Nrf2的核转位及其下游靶蛋白血红素加氧酶1(HO-1)的表达情况。结果与OGD/R模型组比较,不同浓度的FPS均能提高N2a细胞存活率,降低死亡率,且增加Nrf2的核转位及其下游靶蛋白HO-1的表达(P<0.05)。结论 FPS对OGD/R诱导N2a细胞缺血再灌注损伤具有保护作用,其机制可能与激活Nrf2通路有关。

Objective To investigate the effect of Fucoidan （FPS） on N2a cells in rats with ischemia-reperfu- sion injury induced by oxygen-glucose deprivation followed by reperfusion （OGD/R） and its potential molecular mecha- nisms in mice. Methods Ischemia-reperfusion injury models were established in N2a ceils of the mice by OGD for 2 h and followed by reperfusion for 24 h. The experimental cells were randomly divided into control group, OGD/R group and OGD/R ＋ FPS group （ 1, 2, 5 and 10 txg/mL）. Thiazolyl blue tetrazolium bromide （MTT） colorimetric method and lac- tate dehydrogenase （LDH） kit were used to detect the survival rate of cells and release rate of LDH respectively. Western blotting was used to detect the nuclear translocation of Nrf2 and the expression of the downstream target protein heine oxy- genase （ HO-1 ）. Results Compared with those in OGD/R group, OGD/R ＋ FPS group showed significantly higher sur- vival rates of cells, lower death rates in different concentrations of FPS and the increased nuclear translocation of Nrf2 and the expression of the downstream target protein HO-1 （P 〈 0.05）. Conclusion FPS plays an important role in protec- ting the N2a neuroblastoma ceils in mice with ischemic-reperfusion injury induced by OGD/R, and its mechanisms are mainly attributed to the activation of Nrf2 signaling pathways.