LPS对16HBE细胞AQP1、AQP5表达的影响及作用机制

The effects of LPS on AQP1 and AQP5 expression in 16HBE cells and the underline mechanism

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。

This study designed to explore the effect of LPS on the expression of AQP1 and AQP5 in 16 HBE cells and the underline mechanism. 16 HBE cells were cultured in vitro and the m RNA expression of AQP1 and AQP5 was detected by real-time PCR after treatment with LPS at doses of 1,10,100,500 and 1 000 ng/ml for 24 h,or treatment with LPS at 100 ng/ml for 0,3,6,12,24 and 48 h. Then,Western blot was used to test the changes of p38 MAPK signaling pathway after 16 HBE cells were incubated with LPS at 100 ng/ml for 0,5,15,30 and 60 min.Also,16 HBE cells were incubated with LPS at 100 ng/ml with or without SB203580,and then the m RNA and protein expression of AQP1 and AQP5,as well as the changes of p38 MAPK signaling pathway were detected. Data showed that the expression of AQP1 and AQP5 was decreased markedly under the stimulation of LPS in dose- and time-dependent in vitro（P 〈0.01）; the m RNA expression of AQP1 and AQP5 declined significantly after the 16 HBE cells was treated with 100 ng/ml for 3 h,and downed to the bottom at 24 h. LPS could activate p38 MAPK signaling pathway and reach a climax at 30 min. Compared with LPS,LPS＋SB203580 could up-regulate the expressions of AQP1 and AQP5（P 〈0.01）. In conclusion,LPS could down-regulate the expression of AQP1 and AQP5 through activating p38 MAPK signaling pathway in 16 HBE cells.