基于调节Th细胞亚群平衡的清肺口服液抗呼吸道合胞病毒感染的蛋白质组学研究

Respiratory syncytial virus (RSV) pneumonia is a common pediatric disease. Its pathogenic mechanism is not very clear. Modern medical research has revealed that imbalance of Th cell subsets caused by RSV infection will lead to severe immune response syndrome, which is consistent with the "phlegm off the lung" syndrome in traditional Chinese medicine. Our clinical studies have demonstrated that Qingfei oral liquid is an effective drug treatment of children with RSV pneumonia. Besides, our preliminary findings revealed that Qingfei oral liquid can regulate T cell balance and antagonize RSV. A few foreign scholars have reported some specific proteins after RSV infection through proteomics technology, which are associated with cellular antiviral responses, T cell differentiation and balance, and so on. Given the common holistic and systemic features between proteomics research methods and traditional Chinese medicine, this study plans to use new label-free quantitative technologies and multiple bioinformatics analysis to detect differential proteome and related pathways (especially pathways and related cytokines in T cell differentiation and balance) before and after Qingfei oral liquid function both in vitro and in mouse models. This study will reveal the possible pathways and potential targets of Qingfei oral liquid, clarify its function mechanism in RSV antagonism and regulation of T cell balance, promote studies in the therapeutic mechanism of traditional Chinese medicine against RSV pneumonia in children, and provide advanced experimental methodology for clinical treatment and scientific research of viral pneumonia.

呼吸道合胞病毒（RSV）肺炎为儿科常见多发病，其致病机理尚不十分明确。现代医学研究揭示，RSV感染会引起Th细胞亚群失衡，造成严重免疫反应，与中医“痰热闭肺”证候一致。我们的临床研究已证实清肺口服液是治疗RSV肺炎的有效药物，且前期实验研究提示其具有拮抗RSV、调节T细胞平衡的作用。国外有少数学者通过蛋白质组学技术，发现RSV感染前后差异蛋白与抗病毒反应、T细胞分化及平衡等多种功能相关。鉴于蛋白质组学研究方法与中药复方作用都具有整体性、系统性特点，本课题拟基于RSV感染的体外细胞及小鼠模型，首次运用最新非标定量技术及多样化的生物信息学分析，研究清肺口服液作用前后的差异蛋白质组及相关通路、尤其是T细胞分化平衡相关通路及因子，揭示药物可能的作用途径及潜在靶标，阐释其拮抗RSV、调节T细胞平衡的作用机理，促进RSV肺炎的中医药疗效机理研究，为中医药临床治疗及科学研究提供先进的实验方法学。

背景：呼吸道合胞病毒（RSV）肺炎为儿科常见多发病，感染会引起Th细胞亚群失衡，清肺口服液是治疗RSV肺炎的临床有效药物。RSV感染前后差异蛋白与抗病毒反应、T细胞分化及平衡等多种功能相关。本课题利用RSV感染的体外细胞及小鼠模型，研究清肺口服液治疗前后的差异蛋白质组及相关通路，观察治疗后T细胞分化平衡相关通路及炎性因子改变情况。结果发现，清肺口服液可调节呼吸道合胞病毒感染小鼠肺组织中Th1、Th2、Treg和Th17关键转录因子T-bet、GATA3、Foxp3、RORγt及炎性细胞因子IL-12、IL-4、IL-6、IL-17、TNF-α和IL-10的表达，通过STAT1、STAT3和STAT6信号通路，抑制p-STAT1、p-STAT3和p-STAT6的表达水平，调控Th1/Th2和Treg/Th17之间的平衡，从而减轻呼吸道合胞病毒感染小鼠肺组织中的炎症反应炎症，降病理损伤，经功能分类和蛋白互作网络分析，发现清肺口服液可调节细胞骨架重排、免疫反应和代谢变化等发挥抗呼吸道合胞病毒感染作用，明确了清肺口服液抗RSV的部分作用机制，促进RSV肺炎的中医药疗效机理研究，为中医药临床治疗及科学研究提供先进的实验方法学。