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目的探讨吲哚美辛对人羊膜上皮细胞水通道蛋白(Aquaporin,AQP)8表达的影响。方法原代培养人羊膜上皮细胞,采用免疫细胞化学法进行鉴定。将细胞分为实验组和对照组,实验组以不同浓度的吲哚美辛(10、20、50、100、200μmol/L)作用24 h,另以200μmol/L吲哚美辛分别作用6、12、24、36和48 h,以正常培养的细胞为对照,采用RT-PCR和Western blot检测各组细胞AQP 8 mRNA及蛋白的表达变化。结果除10μmol/L作用细胞外,其他浓度的吲哚美辛均可显著下调人羊膜上皮细胞AQP 8 mRNA及蛋白的表达(P<0.01),其中200μmol/L吲哚美辛作用组的表达水平最低;200μmol/L吲哚美辛作用的细胞,从作用12 h开始,AQP 8 mRNA和蛋白的表达水平均显著低于对照组(P<0.01),且呈时间依赖性,48 h时达最低。结论吲哚美辛可下调人羊膜上皮细胞AQP 8 mRNA和蛋白的表达。
Objective To investigate the effect of indomethacin on the expression of aquaporin (AQP) 8 in human amniotic epithelial cells. Methods Primary cultured human amniotic epithelial cells were identified by immunocytochemistry. The cells were divided into experimental group and control group. The experimental groups were treated with indometacin (10, 20, 50, 100, 200μmol / L) for 24 h and indomethacin (200μmol / At 12, 24, 36 and 48 h, the normal cultured cells were used as control. The expression of AQP 8 mRNA and protein in each group was detected by RT-PCR and Western blot. Results All the concentrations of indomethacin significantly down-regulated the expression of AQP 8 mRNA and protein in human amniotic epithelial cells (P <0.01) except for 10 μmol / L-treated cells, and the expression of AQP 8 mRNA and protein in 200 μmol / L indomethacin- (P <0.01). The expression of AQP 8 mRNA and protein in the cells treated with 200 μmol / L indomethacin was significantly lower than that of the control group at 12 h . Conclusion Indometacin can down-regulate the expression of AQP 8 mRNA and protein in human amniotic epithelial cells.