Liang Mingchuan, Meng Yanli, Wang Xiaoxi, Wang Lei, Tang Guixin, Wang Weiming
Heilongjiang Academy of Chinese Medicine Sciences, Harbin, China.
Advanced Microscopy and Instrumentation Research Center, Harbin Institute of Technology, Harbin, China.
Front Mol Biosci. 2022 Jul 15;9:803842. doi: 10.3389/fmolb.2022.803842. eCollection 2022.
Cough is the main symptom of mycoplasma pneumoniae (MP) infection. Cough potential protein transient receptor potential A1 (TRPA1) plays an important role in cough reflex. The purpose of this study was to clarify the mechanism of wogonin, the effective component of Qinbai Qingfei concentrated pellet (Qinbai), in the treatment of cough after MP infection. The Biacore™ system was used to detect whether there was specific binding between Qinbai and cough potential protein TRPA1. Biacore™ fishing technology and UPLC-Q-TOF-MS technology were used during fishing combined active components and identification and analysis of recovered samples. The expression levels of TRPA1, substance P (SP), calcitonin gene-related peptide (CGRP), cough-related proteins, and mRNA in the lung tissues from each group were detected by immunohistochemistry, Western blot, and real-time PCR. Biacore™ results showed that Qinbai had strong specific binding to TRPA1 protein with a binding value of 99.0 resonance unit (RU). The samples obtained from angling were identified and analyzed by UPLC-Q-TOF-MS as wogonin. The results of immunohistochemistry, Western blot, and real-time PCR showed that compared with the model group, the wogonin group had lower expressions of mRNA, TRPA1, SP, and CGRP in the lung tissue of cough mice with MP infection ( < 0.01 or < 0.05), and the effects were superior to those of azithromycin and pentoxyverine control groups. Wogonin can treat cough after MP infection by affecting the expressions of cough-related proteins, such as TRPA1, SP, and CGRP. This study provided a theoretical foundation for the clinical research of Qinbai.
咳嗽是支原体肺炎（MP）感染的主要症状。咳嗽潜在蛋白瞬时受体电位A1（TRPA1）在咳嗽反射中起重要作用。本研究旨在阐明清肺清fei浓缩丸（清肺）的有效成分汉黄芩素治疗MP感染后咳嗽的机制。使用Biacore™系统检测清肺与咳嗽潜在蛋白TRPA1之间是否存在特异性结合。在捕获联合活性成分以及对回收样品进行鉴定和分析的过程中，采用了Biacore™捕获技术和超高效液相色谱-四极杆飞行时间质谱技术。通过免疫组织化学、蛋白质免疫印迹法和实时聚合酶链反应检测各组肺组织中TRPA1、P物质（SP）、降钙素基因相关肽（CGRP）、咳嗽相关蛋白及mRNA的表达水平。Biacore™结果显示，清肺与TRPA1蛋白具有强特异性结合，结合值为99.0共振单位（RU）。通过超高效液相色谱-四极杆飞行时间质谱对垂钓所得样品进行鉴定和分析，确定为汉黄芩素。免疫组织化学、蛋白质免疫印迹法和实时聚合酶链反应结果显示，与模型组相比，汉黄芩素组MP感染咳嗽小鼠肺组织中mRNA、TRPA1、SP和CGRP的表达较低（<0.01或<0.05），且效果优于阿奇霉素和喷托维林对照组。汉黄芩素可通过影响TRPA1、SP和CGRP等咳嗽相关蛋白的表达来治疗MP感染后的咳嗽。本研究为清肺的临床研究提供了理论依据。
