3，4-二羟基苯甲醛作用星形胶质细胞减轻脑微血管内皮细胞和神经元氧糖剥夺/再灌注损伤
3, 4-Dihydroxybenzaldehyde attenuates oxygen-glucose deprivation/reperfusion injury in brain microvascular endothelial cells and neurons by acting on astrocytes
- 上海中医药大学学报 2021年35卷第4期 页码：67-72,100
- 作者机构：
1.云南中医药大学中药学院（云南 昆明 650500）
- 作者简介：
夏霜莉，女，在读硕士生，主要从事中药药理与应用研究
代蓉，副教授，硕士生导师；E-mail：1647029626@qq.com
- 基金信息：云南省科技厅科技计划项目中医联合专项（重点）项目(2017FF117（-004）;2018FF001（-007）);云南省教育厅科学研究基金项目(2020Y0207)
DOI：10.16306/j.1008-861x.2021.04.010
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夏霜莉, 杨媛, 廖陈, 等. 3，4-二羟基苯甲醛作用星形胶质细胞减轻脑微血管内皮细胞和神经元氧糖剥夺/再灌注损伤[J]. 上海中医药大学学报, 2021,35(4):67-72,100.
XIA Shuangli, YANG Yuan, LIAO Chen, et al. 3, 4-Dihydroxybenzaldehyde attenuates oxygen-glucose deprivation/reperfusion injury in brain microvascular endothelial cells and neurons by acting on astrocytes[J]. Academic Journal of Shanghai University of Traditional Chinese Medicine, 2021,35(4):67-72,100.
夏霜莉, 杨媛, 廖陈, 等. 3，4-二羟基苯甲醛作用星形胶质细胞减轻脑微血管内皮细胞和神经元氧糖剥夺/再灌注损伤[J]. 上海中医药大学学报, 2021,35(4):67-72,100. DOI： 10.16306/j.1008-861x.2021.04.010.
XIA Shuangli, YANG Yuan, LIAO Chen, et al. 3, 4-Dihydroxybenzaldehyde attenuates oxygen-glucose deprivation/reperfusion injury in brain microvascular endothelial cells and neurons by acting on astrocytes[J]. Academic Journal of Shanghai University of Traditional Chinese Medicine, 2021,35(4):67-72,100. DOI： 10.16306/j.1008-861x.2021.04.010.
摘要
目的：,2,从神经血管单元（NVU）细胞间相互联系的角度，探讨天麻活性成分3，4-二羟基苯甲醛（3，4-DD）作用星形胶质细胞（Ast）对脑微血管内皮细胞（BMEC）和神经元氧糖剥夺/再灌注（OGD/R）损伤的保护作用。,方法：,2,从乳鼠分离、培养BMEC和Ast。采用Transwell小室分别构建Ast与BMEC、Ast与PC12细胞共培养体系。将共培养细胞分为对照组、模型组、丁苯酞（NBP）干预（1×10,－4,mol/L）组、3，4-DD干预（1×10,－7,mol/L）组。药物组预先给予相应药物干预24 h后，模型组和药物组细胞均诱导OGD/R损伤。采用细胞电阻仪测定各组Transwell小室内侧细胞的跨内皮电阻（TEER）值；试剂盒检测BMEC、PC12细胞乳酸脱氢酶（LDH）活性；ELISA检测培养基中脑源性神经营养因子（BDNF）的含量；PCR检测BMEC细胞闭锁蛋白（,Occludin,）及PC12细胞磷脂酶C-γ（,Plc-γ,）的mRNA表达。,结果：,2,①Ast-BMEC共培养体系：与对照组相比，模型组TEER值、LDH活性、BDNF含量显著降低（,P,<,0.01），,Occludin, mRNA表达显著增加（,P,<,0.01）。与模型组相比，3，4-DD干预组TEER值、LDH活性、BDNF含量以及,Occludin, mRNA表达均显著升高（,P,<,0.01），NBP干预组LDH活性、BDNF含量及,Occludin, mRNA表达亦显著增加（,P,<,0.01）。3，4-DD干预组TEER值较NBP干预组明显升高（,P,<,0.01）。②Ast-PC12共培养体系：与对照组相比，模型组TEER值、LDH活性、BDNF含量显著降低（,P,<,0.01），,Plc-γ, mRNA表达显著增加（,P,<,0.01）。与模型组相比，3，4-DD干预组LDH活性、BDNF含量及,Plc-γ, mRNA表达显著升高（,P,<,0.05，,P,<,0.01），NBP干预组LDH活性及,Plc-γ, mRNA表达亦显著升高（,P,<,0.01）。,结论：,2,3，4-DD通过促进Ast释放BDNF，作用于BMEC和神经元，维持细胞间结构，增强神经元突触可塑性，进而减轻OGD/R损伤。
Abstract
Objective:,2,To investigate the protective effects of 3, 4-dihydroxybenzaldehyde(3, 4-DD), an active component of Gastrodiae Rhizoma, on oxygen-glucose deprivation/reperfusion(OGD/R) injury in brain microvascular endothelial cells(BMEC) and neurons by acting on astrocytes(Ast), from the perspective of interconnection between neurovascular unit(NVU) cells.,Methods:,2,BMEC and Ast were isolated and cultured from neonatal rats.The co-culture systems of Ast and BMEC, Ast and PC12 cells were constructed by Transwell chamber, respectively.The co-culture cells were divided into the control group, model group, butyphthalide(NBP) intervention(1×10,-4,mol/L) group and 3, 4-DD intervention(1×10,-7,mol/L) group.The drug treatment groups were treated with the corresponding drug in advance.After drug intervention for 24 hours, OGD/R injury was induced in the model group and drug treatment groups.The transendothelial electrical resistance(TEER) of the cells inside Transwell chamber was measured by cell electrical resistance meter.The activity of lactate dehydrogenase(LDH) in BMEC and PC12 cells was detected by kit.The content of brain-derived neurotrophic factor(BDNF) in the medium was detected by ELISA.The mRNA expressions of ,Occludin, in BMEC and phospholipase C-γ(,Plc-γ,) in PC12 cells were detected by PCR.,Results:,2,①Ast-BMEC co-culture system: Compared with the control group, the TEER value, LDH activity and BDNF content of the model group were significantly decreased(,P,<,0.01), and the ,Occludin, mRNA expression was significantly increased(,P,<,0.01) .Compared with the model group, the TEER value, LDH activity, BDNF content and ,Occludin, mRNA expression were significantly increased in the 3, 4-DD intervention group(,P,<,0.01), and the LDH activity, BDNF content and ,Occludin, mRNA expression were also significantly increased in the NBP intervention group(,P,<,0.01) .The TEER value of 3, 4-DD intervention group was significantly higher than that of NBP intervention group(,P,<,0.01) .②Ast-PC12 co-culture system: Compared with the control group, the TEER value, LDH activity and BDNF content of the model group were significantly decreased(,P,<,0.01), and the ,Plc-γ, mRNA expression was significantly increased(,P,<,0.01) .Compared with the model group, the LDH activity, BDNF content and ,Plc-γ, mRNA expression were significantly increased in the 3, 4-DD intervention group(,P,<,0.05,P,<,0.01), and the LDH activity and ,Plc-γ, mRNA expression were also significantly increased in the NBP intervention group(,P,<,0.01) .,Conclusion:,2,3, 4-DD can promote the release of BDNF by Ast, act on BMEC and neurons, maintain intercellular structure, enhance synaptic plasticity of neurons, and then alleviate OGD/R injury.
关键词
3，4-二羟基苯甲醛氧糖剥夺/再灌注脑源性神经营养因子星形胶质细胞脑微血管内皮细胞
Keywords
3 4-dihydroxybenzaldehydeoxygen-glucose deprivation/reperfusionbrain-derived neurotrophic factorastrocytebrain microvascular endothelial cell
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