目的探讨脂多糖(LPS)作用于气道上皮细胞后对过氧化物氧化还原酶1(prdx1)表达的影响。方法培养正常人气道上皮细胞株BEAS-2B,以0、1、10mg/LLPS作用于气道上皮细胞12h和24h后,用逆转录-聚合酶链反应(RT—PCR)检测prdx1 mRNA表达;以0、0.1、0.5、1、5、10mg/L LPS作用于气道上皮细胞12h后,用蛋白质免疫印迹试验(Western blotting)检测prdx1蛋白表达。结果RT—PCR结果显示:LPS作用于细胞12h内,随着LPS作用剂量的增加,prdx1 mRNA表达增高,10mg/LLPS作用12hprdx1 mRNA表达较对照组显著增加(2.014±0.197比0.644±0.178,P〈0.05);但随着LPS作用时间的延长,24h时prdx1 mRNA表达有所回落。Western blotting结果显示,随着LPS作用剂量的增加,prdx1蛋白表达逐渐增高,5mg/L LPS作用12h时prdx1蛋白显著高于对照组(1.069±0.175比0.328±0.010,P〈0.05),10mg/LLPS作用12h时维持在高水平(0.984±0.220)。结论10mg/L的LPS作用于BEAS-2B细胞12h后可诱导prdx1的基因和蛋白表达增加。
Objective To investigate the effect of lipopolysaccharide (LPS) on expression of peroxiredoxin 1 (prdxl) in airway epithelial cells. Methods The airway epithelium cell line BEAS-2B was cultivated, and the cells were stimulated with 0, 1, and 10 mg/L of LPS for 12 hours and 24 hours, and then were harvested for prdxl expression detection. The mRNA expression of prdxl was detected by reverse transcription-polymerase chain reaction (RT-PCR). The airway epithelium cells were stimulated with 0, 0.1, 0.5, 1, 5, and 10 mg/L of LPS for 12 hours, and were collected for determination of prdx 1 protein expression by Western blotting. Results RT-PCR results showed that the prdxl mRNA expression was significantly increased within 12 hours of stimulation with elevation of the dosage of LPS. The prdxl mRNA expression at 12 hours of stimulation by 10 mg/L LPS was significantly higher than that in control group (2.014 ± 0.197 vs. 0.644 ± 0.178, P〈0.05). However, with prolongation of LPS stimulation time, the prdxl mRNA expression at 24 hours was slightly declined. Western blotting results showed that the prdxl protein expression was gradually increased with elevation of dosage of LPS. The prdxl protein expression at 12 hours of stimulation with 5 mg/L LPS was significantly higher than that in control group ( 1.069 ± 0.175 vs. 0.328 ± 0.010, P〈0.05), and the expression remained at high level at 12 hours of stimulation with 10 mg/L LPS (0.984 ± 0.220). Conclusion 10 mg/L of LPS can induce the mRNA and protein expression of prdxl in BEAS-2B cell after 12 hours of stimulation.