脂多糖作用于气道上皮细胞后对prdx1表达的影响分析

Analysis of effect of LPS on the expression of prdx1 after airway epithelial cells

目的分析脂多糖(LPS)作用于气道上皮细胞后对prdx1表达的影响。方法在DMEM/F12培养基(含10%胎牛血清)中培养正常人的气道上皮细胞株(BEAS-2B),将培养的细胞收集起来计数后分别在6孔培养板中或培养皿中铺板,待细胞贴壁12 h后再将浓度不同但体积相同的LPS分别作用于气道上皮细胞。将体积相同的磷酸盐缓冲液(PBS)加入对照组。12 h、24 h后收集细胞,提取细胞RNA、蛋白。结果 1 mg/L LPS 12 h组、10 mg/L LPS 12 h组、1mg/L LPS 24 h组、10 mg/L LPS 24 h组气道上皮细胞prdx1 mRNA表达均显著高于对照组(P <0. 05),而在气道上皮细胞prdx1 mRNA表达方面,10 mg/L LPS 24 h组<1 mg/L LPS 24 h组<1 mg/L LPS 12 h组<10 mg/L LPS 12 h组(P <0. 05)。0. 1 mg/L LPS 12 h组、0. 5 mg/L LPS 12 h组、1 mg/L LPS 12 h组、5 mg/L LPS 12 h组、10 mg/L LPS 12 h组气道上皮细胞prdx1蛋白表达均显著高于对照组(P <0. 05),而在气道上皮细胞prdx1蛋白表达方面,0. 1 mg/L LPS 12 h组<0. 5 mg/L LPS 12 h组<1 mg/L LPS 12 h组<10 mg/L LPS 12 h组<5 mg/L LPS 12 h组(P <0. 05)。结论 10 mg/L LPS作用于气道上皮细胞12 h后会提升prdx1基因与蛋白表达。

Objective To analyze the effect of LPS on the expression of prdx1 after airway epithelial cells.Methods Normal human airway epithelial cells(BEAS-2B)were cultured in DMEM/F12 medium(containing 10%fetal bovine serum).The cultured cells were collected and counted.After 12 hours of adherence,LPS with different concentrations but the same volume was added to the airway epithelium,and then the cultured cells were planted in 6-well plate or dish respectively.The same volume of phosphate buffer(PBS)was added to the control group.12 h and 24 h after,the cells were collected and used to extract RNA and protein from cells.Results The expressions of prdx1 mRNA in airway epithelial cells of 1 mg/L LPS 12 h group,10 mg/L LPS 12 h group,1 mg/L LPS 24 h group and 10 mg/L LPS 24 h group were significantly higher than those in the control group(P<0.05),while the expressions of prdx1 mRNA in airway epithelial cells of 10 mg/L LPS 24 h group,1 mg/L LPS 24 h group,1 mg/L LPS 12 h group and 10 mg/L LPS 12 h group were gradually increased(P<0.05).The expressions of prdx1 protein in airway epithelial cells of 0.1 mg/L LPS 12 h group,0.5 mg/L LPS 12 h group,1 mg/L LPS 12 h group,5 mg/L LPS 12 h group,10 mg/L LPS 12 h group were significantly higher than those in the control group(P<0.05).The expressions of prdx1 protein in airway epithelial cells of 0.1 mg/L LPS 12 h group,0.5 mg/L LPS 12 h group,1 mg/L LPS 12 h group,10 mg/L LPS 12 h group and 5 mg/L LPS 12 h group were significantly higher than those in the control group,and gradually increased(P<0.05).Conclusion 10 mg/L LPS can enhance the expressions of prdx1 gene and protein after 12 h in airway epithelial cells.