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目的通过研究乌司他丁对TNF-α作用的A549细胞ENaC-β转录和翻译水平的影响。方法将A549细胞分组为空白组、乌司他丁组、TNF-α组和乌司他丁+TNF-α组,TNF-α浓度为100ng/ml,各组作用时间均为6、12、24h。乌司他丁组及乌司他丁+TNF-α组在12h时间点上分别用1250、2500、5000U/ml3种浓度乌司他丁作用A549细胞,采用浓度5000U/ml乌司他丁分别作用6、24h,RT-PCR半定量检测ENaC-βmRNA表达量,Westernblot半定量检测ENaC-β蛋白浓度。结果①在作用不同时间后,乌司他丁对ENaC-βmRNA和蛋白表达水平的影响与空白组比较差异无统计学意义(P>0.05)。②5000U/ml乌司他丁+TNF-α组分别作用6、12、24h后ENaC-βmRNA及蛋白表达较TNF-α组均明显升高(P<0.01)。③作用12h后,各浓度乌司他丁+TNF-α组ENaC-βmRNA及蛋白表达较TNF-α组均明显升高(P<0.01),且各浓度组间ENaC-βmRNA及蛋白表达差异也有统计学意义(P<0.05)。结论乌司他丁不能直接上调A549细胞ENaC-β转录和翻译水平,但能以浓度依赖方式对抗炎症反应对ENaC-β表达的抑制作用。
Objective To investigate the effects of ulinastatin on the transcription and translation of ENaC-β in A549 cells induced by TNF-α. Methods A549 cells were divided into blank group, ulinastatin group, TNF-α group and ulinastatin + TNF-α group, the concentration of TNF-α was 100ng / ml, and the duration of action was 6,12,24h . Ulinastatin group and ulinastatin + TNF-α group were treated with Ulinastatin at concentration of 1250,2500 and 5000 U / ml for A549 cells respectively at 12h, At 6 and 24 h, the expression of ENaC-β mRNA was detected by RT-PCR and the concentration of ENaC-β was detected by Western blot. Results ① The effect of ulinastatin on the expression of ENaC-β mRNA and protein was not significantly different from that of blank group at different time points (P> 0.05). ② The mRNA and protein expressions of ENaC-β in 5 000 U / ml ulinastatin + TNF-α group were significantly higher than those in TNF-α group at 6, 12 and 24 hours (P <0.01). ③ After 12 hours, the expression of ENaC-βmRNA and protein in ulinastatin + TNF-α group were significantly higher than those in TNF-α group (P <0.01), and the differences of ENaC-βmRNA and protein expression between each concentration group Statistical significance (P <0.05). Conclusion Ulinastatin can not directly up-regulate the transcription and translation of ENaC-β in A549 cells, but it can inhibit the inhibitory effect of inflammatory response on ENaC-β expression in a concentration-dependent manner.