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目的探讨LPS对16HBE细胞AQP1、AQP5表达的影响及作用机制。方法用1、10、100、500、1 000 ng/ml的LPS干预16HBE细胞24 h,real-time PCR检测AQP1、AQP5 m RNA的表达;用100 ng/ml的LPS分别干预16HBE细胞0、3、6、12、24、48 h,real-time PCR检测AQP1、AQP5 m RNA的表达;以100 ng/ml的LPS分别干预16HBE细胞0、5、15、30、60 min,Western blot检测p38 MAPK信号通路的变化;LPS单独或联合p38 MAPK特异性阻断剂SB203580干预16HBE细胞24 h,real-time PCR和Western blot分别检测AQP1、AQP5的变化及p38 MAPK信号通路的变化。结果与对照组相比,10、100、500、1 000 ng/ml的LPS均可显著抑制AQP1和AQP5的表达(P<0.01),且具有剂量依赖效应;100 ng/ml的LPS干预16HBE细胞3 h后,AQP1和AQP5 m RNA的表达即发生下降,并于24 h降到最低;LPS能够显著激活p38 MAPK信号通路,其磷酸化水平于30 min达到高峰;与LPS单独干预组相比,LPS+SB203580组AQP1和AQP5的表达水平发生明显上升(P<0.01)。结论 LPS可通过激活p38 MAPK信号通路下调16HBE细胞AQP1和AQP5的表达。
Objective To investigate the effect of LPS on the expression of AQP1 and AQP5 in 16HBE cells and its mechanism. Methods 16HBE cells were treated with LPS at 1, 10, 100, 500 and 1 000 ng / ml for 24 h, and the expression of AQP1 and AQP5 mRNA was detected by real-time PCR. The cells were treated with 100 ng / ml LPS for 16 hBE cells, The expression of AQP1 and AQP5 mRNA was detected by real-time PCR at 6, 12, 24 and 48 h. The cells were treated with 100 ng / ml LPS for 0, 5, 15, 30 and 60 min respectively. Western blot was used to detect the expression of p38 MAPK The changes of AQP1 and AQP5 and the changes of p38 MAPK signal pathway were detected by real-time PCR and Western blot respectively after intervention with SB203580 or SB203580 alone or in combination with p38 MAPK specific inhibitor SB203580 for 16 h. Results Compared with the control group, LPS at 10, 100, 500 and 1 000 ng / ml significantly inhibited the expression of AQP1 and AQP5 (P <0.01) and in a dose-dependent manner. LPS at 100 ng / ml interfered with 16HBE cells After 3 h, the expression of AQP1 and AQP5 mRNA decreased, and reached the lowest level at 24 h. LPS significantly activated the p38 MAPK signal pathway, and the phosphorylation level peaked at 30 min. Compared with the LPS alone group, The expression of AQP1 and AQP5 in LPS + SB203580 group increased significantly (P <0.01). Conclusion LPS can down-regulate the expression of AQP1 and AQP5 in 16HBE cells by activating the p38 MAPK signaling pathway.