[1]周 爽,王 琰,孙光强,等.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[J].陕西医学杂志,2022,51(4):410-414,418.[doi:DOI:10.3969/j.issn.1000-7377.2022.04.006]
　ZHOU Shuang,WANG Yan,SUN Guangqiang,et al.Effects of overexpression of 5-LOX pathway on proliferation and apoptosis of oxygen-glucose-deprived/reoxygenated microglia[J].,2022,51(4):410-414,418.[doi:DOI:10.3969/j.issn.1000-7377.2022.04.006]
点击复制

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

《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
51
期数:
2022年4期
页码:
410-414,418
栏目:
基础研究
出版日期:
2022-04-05

文章信息/Info

Title:
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; 王 川; 王国全; 李 敏; 王 斌
(陕西中医药大学,陕西 咸阳 712046)
Author(s):
ZHOU Shuang; WANG Yan; SUN Guangqiang; SHI Yongheng; WANG Chuan; WANG Guoquan; LI Min; WANG Bin
(Shaanxi University of Chinese Medicine,Xianyang 712046,China)
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
Keywords:
5-LOX overexpression;  Microglia injury;  Oxygen glucose-deprivation/reoxygenation;  5-LOX pathway;  LTB4 pathway;  CysLTs pathway
分类号:
R 34
DOI:
DOI:10.3969/j.issn.1000-7377.2022.04.006
文献标志码:
A
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。
Abstract:
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.

参考文献/References:

[1] 陈孝男,杨爱琳,赵亚楠,等.缺血性脑中风的发病机制及其常用治疗中药研究进展[J].中国中药杂志,2019,44(3):422-432.
[2] 刘 寒,张欢欢,康亚国,等.黄芩苷、栀子苷配伍对脑缺血大鼠恢复期炎症因子的影响[J].陕西中医药大学学报,2018,41(3):75-81.
[3] Gao Y,Cao X,Zhang X,et al.Brozopine inhibits 15-LOX-2 metabolism pathway after transient focal cerebral ischemia in rats and OGD/R-induced hypoxia injury in PC12 cells[J].Frontiers in Pharmacology,2020,11:99.
[4] Chen Z,Yang J,Zhong J,et al.MicroRNA-193b-3p alleviates focal cerebral ischemia and reperfusion-induced injury in rats by inhibiting 5-lipoxygenase expression[J].Experimental Neurology,2020,327(5):113223.
[5] Liang G,Shi B,Luo W,et al.The protective effect of caffeic acid on global cerebral ischemia-reperfusion injury in rats[J].Behavioral and Brain Functions,2015,11(1):18.
[6] 泰文娇,叶 旋,鲍秀琦,等.小胶质细胞与脑缺血关系的研究进展[J].药学学报,2012,47(3):346-353.
[7] Kong LL,Wang ZY,Hu JF,et al.Inhibition of chemokine-like factor 1 improves blood-brain barrier dysfunction in rats following focal cerebral ischemia[J].Neurosci Lett,2016,627:192-198.
[8] 沈 甜,张 怡,强朝阳,等.黄芩苷栀子苷配伍对脂多糖致BV2小胶质细胞激活损伤的影响[J].世界最新医学信息文摘,2017,17(45):148-150.
[9] 孟凡荣,陈 琛,万海粟,等.慢病毒载体及其研究进展[J].中国肺癌杂志,2014,17(12):870-876.
[10] 李 坤,田余祥,陈海波,等.重组人核糖核酸酶抑制因子 基因的siRNA表达载体构建及B16细胞中基因沉默的鉴定[J].应用与环境生物学报,2007,13(4):519-522.
[11] 袁晓捧,姜希娟,李悠悠,等.小胶质细胞介导的炎性反应参与脑缺血再灌注损伤的研究进展[J].中华老年心脑血管病杂志,2019,21(10):1112-1114.
[12] Murphy RC,Gijón MA.Biosynthesis and metabolism of leukotrienes[J].Biochemical Journal,2007,405(3):379-395.
[13] Yasukuni K,Wataru T,Masamichi O,et al.Leukotriene enhances NMDA-induced inward currents in dorsal horn neurons of the rat spinal cord after peripheral nerve injury[J].Molecular Pain,2015,11:53.
[14] 张晓燕,陈 璐,徐栋敏,等.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[J].浙江大学学报:医学版,2014,43(3):273-280.
[15] Hidekazu T,Masunari S,Masafumi I,et al.A comparative study on the expression of cyclooxygenase and 5-lipoxygenase during cerebral ischemia in humans[J].Acta Neuropathologica,2002,104(6):601-607.
[16] Paul R,Karli WM.Pharmacotherapy of diseases mediated by 5-lipoxygenase pathway eicosanoids[J].Prostaglandins & Other Lipid Mediators,2007,83(3):188-197.
[17] Syed O,Rakesh KS,Salman H,et al.A novel therapeutic potential of cysteinyl leukotrienes and their receptors modulation in the neurological complications associated with Alzheimer's disease[J].European Journal of Pharmacology,2019,842:208-220.
[18] Bhatt L,Roinestad K,Van T,et al.Recent advances in clinical development of leukotriene B4 pathway drugs[J].Semin Immunol,2017,33:65-73.
[19] Richards IM,Griffin RL,Ostveen JA,et al.Effect of the selective leukotriene B4 antagonist U-75302 on antigen-induced bronchopulmonary eosinophilia in sensitized guinea pigs[J].Am Rev Respir Dis,1989,140(6):1712-1716.
[20] Zhang XY,Chen L,Yang Y,et al.Regulation of rotenone-induced microglial activation by 5-lipoxygenase and cysteinyl leukotriene receptor 1[J].Brain Research,2014,1572:59-71.
[21] Tager AM,Luster AD.BLT1 and BLT2:The leukotriene B(4)receptors[J].Prostaglandins Leukot Essent Fatty Acids,2003,69(2-3):123-134.
[22] Davino-Chiovatto JE,Oliveira-Junior MC,Mackenzie B,et al.Montelukast,leukotriene inhibitor,reduces LPS-Induced acute lung inflammation and human neutrophil activation[J].Arch Bronconeumol,2019,55(11):573-580.
[23] 王 月,刘佳俊,王 雷,等.缺血后处理通过升高血清腺苷水平减轻脑缺血再灌注损伤的实验研究[J].陕西医学杂志,2021,50(7):789-792.

备注/Memo

备注/Memo:
基金项目: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)
更新日期/Last Update: 2022-04-07