芪冬活血饮对肺泡上皮细胞钠水转运的研究

Study of Qidong Huoxue Drink on the sodium and water transport in alveolar epithelial cells

目的:研究芪冬活血饮对肺泡上皮细胞A549中钠离子通道α(ENaC-α)和水通道蛋白-1(AQP-1)表达的影响。方法:将细胞随机分为4组:空白组、LPS组、LPS+正常大鼠血清组、LPS+10%药物血清组,用10%芪冬活血饮药物血清培养A549细胞4h后,再加入10μg/mL LPS刺激细胞,RT-PCR法检测肺泡上皮细胞ENaC-α、AQP-1 mRNA表达,Western Blot检测细胞ENaC-α、AQP-1蛋白表达,用ELISA法检测细胞上清液中肿瘤坏死因子α(TNF-α)、白介素-8(IL-8)水平。结果:刺激后,LPS组细胞ENaC-α、AQP-1 mRNA和蛋白表达较空白组降低(P<0.05,P<0.01),细胞上清液TNF-α、IL-8表达显著增高(P<0.05),而10%芪冬活血饮药物血清较LPS组和LPS+正常大鼠血清组则能促进ENaC-α、AQP-1 mRNA和蛋白表达(P<0.05,P<0.01),降低TNF-α、IL-8释放(P<0.05)。结论:芪冬活血饮能减轻LPS诱导下肺泡上皮细胞的炎症损伤,恢复肺泡上皮细胞正常的钠水转运,从而减轻急性肺损伤时肺水肿程度。

Objective:To study the effect of Qidong Huoxue Drink on the expression of ENaC-αand AQP-1 in alveolar epithelial cells(A549).Methods:Cells were randomly divided into four groups:blank group,LPS group,LPS+normal serum group,LPS+10%drug serum group.A549 cells were cultured with 10%Qidong Huoxue Drink drug serum for 4 hours,then stimulated by 10μg/m L LPS.The mRNA expressions of ENaC-αand AQP-1 were detected by RT-PCR,protein of ENaC-αand AQP-1 were detected by Western Blot and the TNF-α、IL-8 in supernatant of A549 cells were detected by ELISA.Results:The mRNA and protein expression of ENaC-αand AQP-1 were decreased(P<0.05,P<0.01),while the secretion of TNF-αand IL-8 in cell supernatant were significantly increased(P<0.05),when with LPS group compared with the blank group.But when the cells were pretreated with the 10%Qidong Huoxue Drink drug serum,the expression of ENaC-αand AQP-1 was promoted by Qidong Huoxue Drink drug serum(P<0.05,P<0.01),and the secretion and release of TNF-αand IL-8 were decreased compared with the LPS group and LPS+normal serum group(P<0.05).Conclusion:Qidong Huoxue Drink could alleviate LPS-induced inflammation injury of alveolar epithelial cells,restore normal sodium and water transport of alveolar epithelial cells,and reduce the occurrence of pulmonary edema.