本文選題：中暑　切入點(diǎn)：肺微血管內(nèi)皮細(xì)胞　出處：《解放軍醫(yī)學(xué)雜志》2017年04期

【摘要】：目的研究絲裂原活化蛋白激酶(MAPKs)活化對(duì)熱打擊致小鼠肺微血管內(nèi)皮細(xì)胞(PMVECs)凋亡的影響。方法建立重癥中暑小鼠模型,采用TUNEL染色及免疫組化檢測(cè)肺組織損傷情況。二次磁珠分選法分離乳鼠PMVECs,TUNEL染色檢測(cè)PMVECs凋亡情況,Western blotting檢測(cè)熱打擊恢復(fù)期(0、2、6h)MAPKs家族活化情況。通過檢測(cè)單層內(nèi)皮細(xì)胞跨膜電阻(TEER)及辣根過氧化物酶(HRP)值觀察不同熱打擊溫度對(duì)單層細(xì)胞通透性的影響,同時(shí)使用MAPKs家族抑制劑檢測(cè)熱打擊對(duì)單層細(xì)胞通透性及凋亡的影響。結(jié)果在重癥中暑小鼠恢復(fù)期肺組織中可觀察到PMVECs發(fā)生凋亡。TUNEL染色發(fā)現(xiàn)隨著恢復(fù)期時(shí)間的延長,PMVECs凋亡數(shù)目增多,熱打擊可使PMVECs MAPKs家族活化且微血管通透性增加,給予p38活化抑制劑SB203580及ERK活化抑制劑PD98059預(yù)處理后細(xì)胞通透性增加,凋亡數(shù)目增多,而給予JNK抑制劑SP600125預(yù)處理后細(xì)胞則出現(xiàn)相反的變化。結(jié)論重癥中暑小鼠PMVECs可發(fā)生凋亡,p38及ERK起著抗凋亡的作用,JNK起著促凋亡的作用。
[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.
【作者單位】： 廣州軍區(qū)廣州總醫(yī)院重癥醫(yī)學(xué)科;南方醫(yī)科大學(xué)南方醫(yī)院重癥醫(yī)學(xué)科;廣州中醫(yī)藥大學(xué)研究生院;
【基金】：國家自然科學(xué)基金(81671896,81471839) 南方醫(yī)科大學(xué)南方醫(yī)院院長基金(2016C016)~~
【分類號(hào)】：R594.12
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本文編號(hào)：1706803