基础医学论著

SphK2在氧糖剥夺/复糖复氧模型中对BV-2细胞损伤的影响*

  • 张佳乐 ,
  • 李家任 ,
  • 张雅轩 ,
  • 李超然 ,
  • 乔婧 ,
  • 姚璐 ,
  • 牛愉捷 ,
  • 郝肖琼
展开
  • 1.包头医学院基础医学与法医学院,内蒙古包头 014040;
    2.包头医学院 2020级研究生
郝肖琼

收稿日期: 2023-06-14

  网络出版日期: 2024-11-19

基金资助

* 包头医学院科研基金项目青苗计划(BYJJ-QM201905);包头医学院博士科研启动基金(BSJJ201806);包头医学院 2021年度花蕾计划项目(HLBTMC202247)

Effect of SphK2 on BV-2 cell injury in oxygen    glucose deprivation/reoxygenation models

  • ZHANG Jiale ,
  • LI Jiaren ,
  • ZHANG Yaxuan ,
  • LI Chaoran ,
  • QIAO Jing ,
  • YAO Lu ,
  • NIU Yujie ,
  • HAO Xiaoqiong
Expand
  • 1. School of Basic Medicine and Forensic Medicine, Baotou Medical College, Baotou 014040,China;
    2. The 2020 Graduate Student, Baotou Medical College

Received date: 2023-06-14

  Online published: 2024-11-19

摘要

目的:观察BV-2小胶质细胞在氧糖剥夺/复糖复氧(OGD/R)中鞘氨醇激酶2(SphK2)的表达情况,并探讨SphK2是否参与了OGD/R致BV-2小胶质细胞的损伤过程。方法:采用小鼠BV-2小胶质细胞制备OGD/R模型,选取氧糖剥夺时间为2 h,复氧复糖时间为0 h,3 h,6 h,12 h,24 h。(1)设对照组,OGD 2 h组,OGD 2 h/R 3 h组,OGD 2 h/R 6 h组,OGD 2 h/R 12 h组,OGD 2 h/R 24 h组,观察细胞形态变化。细胞经OGD/R处理后CCK-8法检测细胞存活率。(2)设对照组,OGD 2 h/R 12 h组,OGD 2 h/R 12 h+ABC294640(SphK2特异抑制剂)组,用q-PCR及Western blot测定SphK2的基因及蛋白表达变化。结果:与对照组相比,随着OGD/R时间的延长,BV-2活力逐渐下降(P<0.001),细胞形态发生了不同程度的改变,且有不同程度的损伤;通过观察细胞生存情况确定OGD2 h/R12 h为最适宜的OGD/R时间,与对照组相比,经OGD/R处理后BV-2细胞中SphK2 mRNA表达水平显著升高(P<0.05),加入ABC294640显著抑制其表达升高(P<0.001);与对照组相比,经OGD/R处理后BV-2细胞中SphK2蛋白表达水平显著升高(P<0.001),加入ABC294640后,SphK2蛋白表达明显降低(P<0.001)。结论:OGD/R损伤后的小胶质细胞可以被SphK2激活,在其损伤过程中发挥着调控作用。

本文引用格式

张佳乐 , 李家任 , 张雅轩 , 李超然 , 乔婧 , 姚璐 , 牛愉捷 , 郝肖琼 . SphK2在氧糖剥夺/复糖复氧模型中对BV-2细胞损伤的影响*[J]. 包头医学院学报, 2024 , 40(9) : 24 -28 . DOI: 10.16833/j.cnki.jbmc.2024.09.005

Abstract

Objective: To investigate the expression of sphingosine kinase 2 (SphK2) in BV-2 microglia during oxygen glucose deprivation/reoxygenation (OGD/R), and to investigate whether SphK2 is involved in the damage process of BV-2 microglia induced by OGD/R. Methods: The OGD/R model was established using mouse BV-2 microglial cells. The oxygen glucose deprivation time was 2 hours, and the reoxygenation and rehydration time was 0, 3, 6, 12, and 24 hours. (1) The control group, OGD2h group, OGD2h/R3h group, OGD2h/R6h group, OGD2h/R12h group and OGD2h/R24h group were set up to observe the changes of cell morphology. Cell viability was detected by CCK-8 method after OGD/R treatment; (2) The control group, OGD2h/ R12h group and OGD2h/R12h+ABC294640 (SphK2 specific inhibitor) group were set up, and the expression of SphK2 gene and protein was determined by q-PCR and Western blot. Results: Compared with the control group, with the prolongation of OGD/R time, the activity of BV-2 gradually decreased (P<0.001), and the cell morphology changed to varying degrees, with varying degrees of injury By observing cell survival, OGD2h/R12h was determined as the most suitable OGD/R time. Compared with the control group, the expression level of Sphk2 mRNA in BV-2 cells treated with OGD/R was significantly increased (P<0.05), and the addition of ABC294640 significantly inhibited the increase in its expression (P<0.001). Compared with the control group, the expression level of SphK2 protein in BV-2 cells treated with OGD/R was significantly increased (P<0.001), while the expression of SphK2 protein was significantly decreased after adding ABC294640 (P<0.001). Conclusion: SphK2 activates microglia after OGD/R injury and plays a regulatory role in the process of OGD/R injury.

参考文献

[1] Zeng SL, Zhao Z, Zheng SN, et al.The E3 ubiquitin ligase TRIM31 is involved in cerebral ischemic injury by promoting degradation of TIGAR[J].Redox Biol, 2021,45:102058.
[2] Zhu H, Hu SP, Li YT, et al.Interleukins and ischemic stroke[J].Front Immunol, 2022,13:828447.
[3] Zheng K, Lin LM, Jiang W, et al.Single-cell RNA-seq reveals the transcriptional landscape in ischemic stroke[J].J Cereb Blood Flow Metab, 2022,42(1):56-73.
[4] Rami A, Bechmann I, Stehlej H.Exploiting endogenous anti-apoptotic proteins for novel therapeutic strategies in cerebral ischemia[J].Prog Neurobiol, 2008,85(3):273-296.
[5] 梁晓瑜,郑丽芳.小胶质细胞表型转换在脑缺血再灌注中作用的研究进展[J].深圳中西医结合杂志,2022,32(14):133-136.
[6] Peng L, Hu GQ, Yao QF, et al.Microglia autophagy in ischemic stroke: a double-edged sword[J].Front Immunol, 2022,13:1013311.
[7] Qin C, Yang S, Chuy H, et al.Signaling pathways involved in ischemic stroke: molecular mechanisms and therapeutic interventions[J].Signal Transduct Target Ther, 2022,7(1):215.
[8] Li CZ, Wu BH, Li YS, et al.Loss of sphingosine kinase 2 promotes the expansion of hematopoietic stem cells by improving their metabolic fitness[J].Blood, 2022,140(15):1686-1701.
[9] 韦曦华,王泽群,陈靖京,等.鞘氨醇激酶和1-磷酸鞘氨醇及其受体信号在肿瘤微环境中的研究进展[J].药学学报,2023,58(3):571-580.
[10] 张金淼,郝清静,江凯旋,等.鞘氨醇激酶2在肿瘤中的作用及其抑制剂研究进展[J].药学学报,2020,55(9):2062-2069.
[11] Zhangs Q, Xiao J, Chen M, et al.Sphingosine-1-phosphate signaling in ischemic stroke: from bench to bedside and beyond[J].Front Cell Neurosci, 2021,15:781098.
[12] Cheng X, Yang YL, Liw H, et al.Dynamic alterations of brain injury, functional recovery, and metabolites profile after cerebral ischemia/reperfusion in rats contributes to potential biomarkers[J].J Mol Neurosci, 2020,70(5):667-676.
[13] 熊娟,庞月珊,吴忧,等.氧糖剥夺/再灌注损伤小胶质细胞通过上调胸腺素β4影响PI3K/AKT信号通路[J].西部医学,2022,34(12):1766-1771.
[14] 高燕,付旭阳,贺帅,等.瓜子金皂苷己通过SOCS3抑制氧糖剥夺/复氧所致小胶质细胞的炎症反应[J].包头医学院学报,2021,37(4):64-68.
[15] Olajideo A, Iwuanyanwuv U, Adegbolao D, et al.SARS-CoV-2 spike glycoprotein S1 induces neuroinflammation in BV-2 microglia[J].Mol Neurobiol, 2022,59(1):445-458.
[16] Liu XL, Zhang MM, Liu HN, et al.Bone marrow mesenchymal stem cell-derived exosomes attenuate cerebral ischemia-reperfusion injury-induced neuroinflammation and pyroptosis by modulating microglia M1/M2 phenotypes[J].Exp Neurol, 2021,341:113700.
[17] 刘璞,高飞.鞘氨醇激酶2与脑相关性疾病的研究现状[J].医学信息,2021,34(3):46-48,55.
[18] Zeng YY, Zhang W, Xue TT, et al.Sphk1-induced autophagy in microglia promotes neuronal injury following cerebral ischaemia-reperfusion[J].Eur J Neurosci, 2022,56(4):4287-4303.
文章导航

/