论文部分内容阅读
目的:观察香烟烟雾提取(CSE)诱导人类肺泡Ⅱ型上皮细胞A549后I-κBα的变化及甲基强的松龙(Mp)对其的影响。方法:体外培养A549细胞系,根据条件不同将其分为3组:(1)对照组:无血清DMEM处理。(2)CSE组:10%浓度CSE处理。(3)CSE+Mp组:CSE刺激后加入0.015%甲基强的松龙处理。于不同时间段收集各组细胞裂解物后,应用免疫印迹法(Western blot)、酶联免疫法(ELISA)观察I-κBα的变化。结果:对照组无变化,CSE组I-κBα从处理后15 min开始后减少,30 min后消失,CSE+Mp组I-κBα在15~30 min内始终有本底表达,3组间I-κBα蛋白的表达水平差异有统计学意义(P<0.01)。结论:甲基强的松龙能使CSE刺激后A549细胞I-κBα蛋白表达增加。
OBJECTIVE: To observe the changes of I-κBα and the effect of methylprednisolone (Mp) on the A549 cells induced by cigarette smoke extraction (CSE) in human alveolar type Ⅱ epithelial cells. Methods: The A549 cell line was cultured in vitro and divided into three groups according to different conditions: (1) control group: serum-free DMEM. (2) CSE group: 10% CSE-treated. (3) CSE + Mp group: CSE stimulated by adding 0.015% methylprednisolone treatment. After collecting the cell lysates at different time points, the changes of I-κBα were observed by Western blot and enzyme-linked immunosorbent assay (ELISA). Results: There was no change in the control group. The I-κBα in CSE group decreased from 15 min after treatment and disappeared after 30 min. The expression of I-κBα in CSE + Mp group was always background at 15-30 min. κBα protein expression was significantly different (P <0.01). CONCLUSION: Methylprednisolone can increase the expression of I-κBα protein in A549 cells after CSE stimulation.