金水六君煎及其拆方含药血清对A549细胞中黏蛋白MUC5AC及水通道蛋白AQP5表达的影响

目的 观察金水六君煎及其拆方含药血清对肺腺癌细胞A549黏液高分泌模型黏蛋白5AC（MUC5AC）及水通道蛋白5（AQP5）表达的影响，探讨金水六君煎治疗气道黏液高分泌的作用机制。方法 将A549细胞分为空白组、模型组、金水六君煎组、养阴组、化痰组。采用人中性粒弹性蛋白酶（HNE）25 nmmol/L诱导A549细胞黏液高分泌，20%金水六君煎及其拆方含药血清干预24 h。RT-PCR、Western blot法检测细胞MUC5AC、AQP5基因与蛋白表达。ELISA法检测细胞上清液MUC5AC、AQP5蛋白表达。结果 与空白组比较，模型组细胞MUC5AC mRNA与蛋白表达明显增多（P<0.05），AQP5 mRNA与蛋白表达明显降低（P<0.05）。与模型组比较，金水六君煎组及养阴组细胞MUC5AC mRNA表达明显降低（P<0.05），AQP5 mRNA表达明显增多（P<0.05）；化痰组A549细胞MUC5AC蛋白表达显著降低（P<0.01），AQP5蛋白表达明显升高（P<0.05）。模型组细胞上清液MUC5AC、AQP5蛋白表达较空白组差异无统计学意义（P>0.05）。结论 金水六君煎可明显抑制A549细胞MUC5AC产生，促进AQP5表达，纠正黏蛋白/水盐比例失衡可能是其治疗气道黏液高分泌的可能机制。

Effects of Serum Containing Jinshui Liujun Decoction and Its Components on the Expression of Mucin MUC5AC and Aquaporin AQP5 in A549 Cell Mucus Hypersecretion Model

Objective To investigate the effects of Jinshui Liujun Decoction and its components on the expression of mucin 5AC (MUC5AC) and aquaporin 5 (AQP5) in the mucus hypersecretion model of human lung adenocarcinoma A549 cells, and to explore the mechanism of action of Jinshui Liujun Decoction in the treatment of airway mucus hypersecretion. Methods A549 cells were divided into blank group, model group, Jinshui Liujun Decoction group, Yin-nourishing group, and phlegm-resolving group. Mucus hypersecretion in A549 cells was induced using human neutral elastase (HNE) at a concentration of 25 nmmol/L, followed by intervention with 20% serum containing Jinshui Liujun Decoction and its components for 24 h. RT-PCR and Western blot were used to determine the mRNA and protein expression levels of MUC5AC and AQP5. Meanwhile, enzyme-linked immunosorbent assay was used to measure the expression of MUC5AC and AQP5 in cell supernatant. Results Compared with the blank group, the model group had significantly increased expression of MUC5AC mRNA and protein (P<0.05) and significantly reduced expression of AQP5 mRNA and protein (P<0.05). Compared with the model group, the Jinshui Liujun Decoction group and the Yin-nourishing group had significantly reduced expression of MUC5AC mRNA (P<0.05) and significantly increased expression of AQP5 mRNA (P<0.05). The phlegm-revolving group showed a significant reduction in the expression of MUC5AC protein (P<0.01) and a significant increase in the expression of AQP5 protein (P<0.05). There was no significant difference in the protein expression of MUC5AC and AQP5 in cell supernatant between the model group and the blank group (P>0.05). Conclusion Jinshui Liujun Decoction can significantly inhibit the expression of MUC5AC and promote the expression of AQP5 in A549 cells. Its ability to correct the imbalance of mucin/water-salt ratio may be a possible mechanism for the treatment of airway mucus hypersecretion.