止嗽散调控黏蛋白祛痰作用研究

Study on the expectorant effect of Zhisou Powder on regulating mucin

关键词:  止嗽散  NCI-H292细胞  水通道蛋白1  水通道蛋白5  黏蛋白5AC

Key words:  Zhisou Powder  NCI-H292 cell  aquaporin 1  aquaporin 5  mucin 5AC

摘要:

[目的] 寻找止嗽散的研究热点，并揭示其对中性粒细胞弹性蛋白酶（NE）诱导的气道水通道蛋白1（AQP1）、水通道蛋白5（AQP5）、黏蛋白5AC（MUC5AC）的调控作用。[方法] 将NCI-H292细胞分为对照组、NE组和止嗽散组，使用实时荧光定量聚合酶链反应（qPCR）方法对AQP1 mRNA、AQP5 mRNA、MUC5AC mRNA含量进行检测，使用蛋白免疫印迹（Western Blot）方法对AQP1、AQP5蛋白进行检测，使用酶联免疫吸附（ELISA）方法对MUC5AC蛋白进行检测。[结果] 1）研究热点：针对病种为咳嗽、慢性咳嗽、外感咳嗽，中医特色为辨证论治。2）NE诱导的NCI-H292。与对照组比较，NE组AQP1 mRNA、AQP5 mRNA表达量下降，MUC5AC mRNA表达量升高，AQP5蛋白表达水平降低，MUC5AC蛋白升高，AQP1蛋白变化不明显。与NE组比较，止嗽散治疗后MUC5AC mRNA明显下降，MUC5AC有下降趋势，AQP1、AQP5变化不明显。[结论] 止嗽散可以下调NCI-H292细胞被NE诱导后高表达的MUC5AC mRNA，对高表达的MUC5AC蛋白存在下调趋势。提示止嗽散发挥祛痰功效的作用机制可能主要与黏蛋白调控有关，但相关结论还需要进一步通过临床研究与动物实验进行验证。

Abstract:

[Objective] Looking for research hotspots in Zhisou Powder（ZSP）. To study the regulatory effect of ZSP on airway aquaporin（AQP） 1，5 and mucin（MUC） 5AC induced by neutrophil elastase（NE）. [Methods] NCI-H292 cells were divided into control group，NE group，and ZSP intervention group. The content of AQP1 mRNA，AQP5 mRNA，and MUC5AC mRNA was detected using qPCR method，Western blot method was used to detect AQP1 and AQP5 proteins，and ELISA method was used to detect MUC5AC protein. [Result] 1） Research hotspot disease：cough，chronic cough，and exogenous cough，the characteristics of traditional Chinese medicine is syndrome differentiation. 2） NE induced NCI-H292：compared with the control group，the NE group showed a decrease in AQP1 mRNA and AQP5 mRNA，an increase in MUC5AC mRNA，a decrease in AQP5 protein expression， an increase in MUC5AC protein，and no significant changes in AQP1 protein. Compared with the NE group， after treatment with ZSP，MUC5AC mRNA significantly decreased，with a downward trend in MUC5AC，and no significant changes in AQP1 and AQP5. [Conclusion] ZSP can downregulate the high expression of MUC5AC mRNA in NCI-H292 cells induced by NE，and has a downward trend towards the high expression of MUC5AC protein. This suggests that the mechanism of action of the anti-cough，volatile and expectorant effects may be mainly related to the regulation of mucin，but the relevant conclusions still need to be further validated through clinical and animal experiments.