C-8神经酰胺对肺泡上皮细胞通透性调节作用的研究

Regulating effect of C8-ceramide on alveolar epithelial barrier cell permeability

目的探讨C-8神经酰胺对肺泡上皮细胞通透性的调节作用。方法体外分离培养的胎鼠肺泡Ⅱ型上皮细胞(alveolar type II epithelial cell,AECⅡ)在Transwell上构建成单层细胞,通过不同浓度的C-8神经酰胺(0,50,100μmol/L)刺激,并测定跨上皮电阻值(TEER);分别应用免疫印迹检测凋亡相关蛋白cleaved-caspase 3、流式细胞技术检测细胞凋亡情况。结果不同浓度的C-8神经酰胺刺激AECII单层24 h后,TEER值明显下降;C-8神经酰胺作用后,免疫印迹检测表明cleaved-caspase 3表达升高,而凋亡率明显升高。结论 C-8神经酰胺可能通过促进肺泡上皮细胞凋亡,增加肺泡上皮屏障通透性。

Objective To investigate the regulating effect of C8-ceramide on alveolar epithelial cell permeability. Methods The primarily cultured AEC Ⅱ were seeded on Transwell polyester membranes to construct in vitro models of monolayers. They were stimulated with different concentrations of C8-ceramide （0, 50, 100 μmol/L） for 24 hours, and the monolayer permeability was measured by TEER. Then, the annexin V externalization assay was performed for apoptosis, and immunoblotting for caspase-3 expression. Results After the stimulation with C8-ceramide （0, 50, 100 μmol/L） for 24 hours, the value pf TEER decreased significantly, and the expression of cleaved-caspase 3 and the apoptotic rate increased significantly. Conclusion C8-ceramide may regulate the alveolar epithelial cell permeability by increasing AEC II apoptosis.