蛋白酪氨酸激酶6对TNF-α诱导的人气道上皮屏障功能减损的影响

发布时间：2019-06-14 14:57

【摘要】：目的:探讨蛋白酪氨酸激酶6(PTK6)对TNF-α诱导的人气道上皮屏障功能减损的影响及相关机制。方法:体外培养人气道上皮16HBE细胞,予TNF-α刺激,分别转染PTK6 siRNA和recombined PTK6,以转染Scramble siRNA和Empty vector为对照。四甲基偶氮唑盐法(MTT)检测细胞活性;跨皮细胞组抗仪检测细胞跨皮细胞阻抗(TER);辣根过氧化物酶(HRP)流量法反应细胞通透性;RT-PCR检测ZO-1、Occludin mRNA水平;Western blot检测PTK6、ZO-1、Occludin、磷酸化细胞外信号调节激酶1/2(p-ERK1/2)、磷酸化c-Jun氨基端激酶1/2(p-JNK1/2)和磷酸化p38(p-p38)蛋白水平。结果:TNF-α刺激后,细胞ZO-1、Occludin转录及蛋白水平、TER值显著降低,细胞通透性增高,同时PTK6、p-ERK1/2、p-JNK1/2和p-p38蛋白水平显著增高(P0.05);上调PTK6使ZO-1、Occludin转录及蛋白水平和TER值进一步降低,细胞通透性和p-JNK1/2、p-p38蛋白水平也进一步增高(P0.05),但下调PTK6后上述指标呈相反变化(P0.05);上调或下调PTK6对p-ERK1/2无明显影响。结论:下调PTK6可以降低JNK1/2、p38MAPK磷酸化水平,进而改善TNF-α诱导的气道上皮屏障功能减损。
[Abstract]:Aim: to investigate the effect of protein tyrosine kinase 6 (PTK6) on TNF- 伪 induced impairment of human airway epithelial barrier function and its related mechanism. Methods: human airway epithelial 16HBE cells were cultured in vitro and stimulated by TNF- 伪. PTK6 siRNA and recombined PTK6, were transfected with Scramble siRNA and Empty vector as control. The cell activity was detected by (MTT) assay, the cell permeability was detected by (TER); horseradish peroxide (HRP) flow assay, and the level of ZO-1,Occludin mRNA was detected by RT-PCR. The levels of extracellular signal-regulated kinase 1 鈮，

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