过表达5-脂氧合酶通路对氧糖剥夺/复氧小胶质细胞增殖、凋亡的影响  

Effects of overexpression of 5-LOX pathway on proliferation and apoptosis of oxygen-glucose-deprived/reoxygenated microglia

作　　者：周爽 王琰 孙光强 史永恒[1] 王川[1] 王国全[1] 李敏[1] 王斌[1] ZHOU Shuang;WANG Yan;SUN Guangqiang;SHI Yongheng;WANG Chuan;WANG Guoquan;LI Min;WANG Bin(Shaanxi University of Chinese Medicine,Xianyang 712046,China)

机构地区：[1]陕西中医药大学,陕西咸阳712046

出　　处：《陕西医学杂志》2022年第4期410-414,418,共6页Shaanxi Medical Journal

基　　金：陕西省中医药优秀中青年科技骨干人才队伍建设项目(2);陕西中医药大学学科创新团队(2019-YL13)。

摘　　要：目的:探索5-脂氧合酶(5-LOX)通路在脑缺血炎性损伤中发挥神经保护作用机制。方法:通过慢病毒包装构建5-LOX过表达载体病毒,转染BV2细胞,建立5-LOX过表达的氧糖剥夺/复氧(OGD/R)BV2细胞模型。OGD/R BV2细胞分别给予抑制剂齐留通(Zileuton)、半胱氨酰白三烯受体阻断剂孟鲁司特(Montelukast)和白三烯B4受体抑制剂U75302,分别阻断5-LOX、白三烯B4受体(BLTs)和半胱氨酰白三烯受体(CysLTRs)。实验分为空载体对照组、过表达组、模型组和给药组,显微镜观察细胞形态变化,采用MTT与流式细胞术分别检测空载体对照组与5-LOX过表达组细胞活力与凋亡。结果:显微镜观察显示出空载体对照组OGD后细胞数目减少,细胞形态逐渐形成“阿米巴状”,5-LOX过表达增加细胞损伤;与模型组相比,给予Zileuton、Montelukast、U75302后,细胞形态恢复,减缓细胞的“阿米巴状”。BV2细胞缺氧缺糖4 h复氧12 h后,与空载体对照组相比,过表达5-LOX细胞活力下降(P<0.01),细胞凋亡增加(P<0.01);与模型组相比,过表达5-LOX介导OGD/R损伤;Zileuton、Montelukast、U75302显著提高细胞活力(均P<0.01),减少细胞凋亡(P<0.01)。结论:5-LOX通过CysLTs/CysLTRs和LTB4/BLTs途径介导小胶质细胞OGD/R的损伤和激活,这一结果能够为脑缺血/再灌注损伤的诊断和临床治疗提供分子基础。Objective:To explore the neuroprotective mechanism of 5-lipoxygenase(5-LOX)pathway in cerebral ischemia inflammatory injury.Methods:5-LOX overexpressed vector virus was constructed by lentivirus packaging,and transfected BV2 cells to establish 5-LOX overexpressed oxygen-glucose-deprived/reoxygenated(OGD/R)BV2 cell model.OGD/R BV2 cells were treated with the inhibitor zileuton,the cysteinyl leukotriene receptor(CysLTRs)blocker montelukast and the leukotriene B4 receptor(BLTs)inhibitor U75302 to block 5-LOX,BLTs and CysLTRs,respectively.They were divided into empty vector control group,overexpression group,model group and administration group.The morphological changes of cells were observed by microscopy.MTT and flow cytometry were used to detect cell viability and apoptosis in the empty vector control group and 5-LOX overexpression group.Results:Microscopic observation showed that the number of cells in the empty vector control group decreased after OGD,and the cell morphology gradually formed amoeba shape.Overexpression of 5-LOX increased cell damage.Compared with the model group,the cell morphology recovered after administration of zileuton,montelukast and U75302.BV2 cells were oxygen-glucose-deprived for 4 hours and reoxygenated for 12 hours,compared with the empty vector control group,the viability of cells overexpressing 5-LOX decreased and apoptosis increased(all P<0.01);compared with the model group,overexpressing 5-LOX mediated OGD/R injury;zileuton,montelukast and U75302 significantly increased cell viability and decreased apoptosis(all P<0.01).Conclusion:5-LOX mediates the injury and activation of microglia OGD/R through CysLTs/CysLTRs and LTB4/BLTs pathways,which can provide a molecular basis for the diagnosis and clinical treatment of cerebral ischemia/reperfusion injury.

关 键 词：5-LOX过表达 小胶质细胞损伤 氧糖剥夺/复氧 5-LOX通路 LTB4通路 CysLTs通路 

分 类 号：R34[医药卫生—基础医学]