MAPKs家族在中暑小鼠肺微血管内皮细胞凋亡中的作用及机制研究

发布时间：2018-04-03 20:34

  本文选题：中暑　切入点：肺微血管内皮细胞　出处：《解放军医学杂志》2017年04期

【摘要】：目的研究丝裂原活化蛋白激酶(MAPKs)活化对热打击致小鼠肺微血管内皮细胞(PMVECs)凋亡的影响。方法建立重症中暑小鼠模型,采用TUNEL染色及免疫组化检测肺组织损伤情况。二次磁珠分选法分离乳鼠PMVECs,TUNEL染色检测PMVECs凋亡情况,Western blotting检测热打击恢复期(0、2、6h)MAPKs家族活化情况。通过检测单层内皮细胞跨膜电阻(TEER)及辣根过氧化物酶(HRP)值观察不同热打击温度对单层细胞通透性的影响,同时使用MAPKs家族抑制剂检测热打击对单层细胞通透性及凋亡的影响。结果在重症中暑小鼠恢复期肺组织中可观察到PMVECs发生凋亡。TUNEL染色发现随着恢复期时间的延长,PMVECs凋亡数目增多,热打击可使PMVECs MAPKs家族活化且微血管通透性增加,给予p38活化抑制剂SB203580及ERK活化抑制剂PD98059预处理后细胞通透性增加,凋亡数目增多,而给予JNK抑制剂SP600125预处理后细胞则出现相反的变化。结论重症中暑小鼠PMVECs可发生凋亡,p38及ERK起着抗凋亡的作用,JNK起着促凋亡的作用。
[Abstract]:Objective to study the effect of mitogen-activated protein kinase (MAPKs) activation on heat shock induced apoptosis of mouse pulmonary microvascular endothelial cells (PMVECs).Methods the model of severe heat stroke in mice was established and lung injury was detected by TUNEL staining and immunohistochemistry.The apoptosis of PMVECs was detected by secondary magnetic bead sorting method and the activation of MAPKs family was detected by Western blotting.The effects of heat shock temperature on the permeability of monolayer cells were observed by measuring the values of transmembrane resistance and horseradish peroxidase (HRP) of monolayer endothelial cells. Meanwhile, the effects of thermal shock on the permeability and apoptosis of monolayer cells were detected by MAPKs family inhibitors.After pretreatment with p38 activation inhibitor SB203580 and ERK activation inhibitor PD98059, the cell permeability increased and the number of apoptosis increased. However, the cells pretreated with JNK inhibitor SP600125 showed the opposite change.Conclusion PMVECs can induce apoptosis in severe heatstroke mice. P38 and ERK play an anti-apoptotic role.
【作者单位】： 广州军区广州总医院重症医学科;南方医科大学南方医院重症医学科;广州中医药大学研究生院;
【基金】：国家自然科学基金(81671896,81471839) 南方医科大学南方医院院长基金(2016C016)~~
【分类号】：R594.12
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本文编号：1706803