清肺口服液对呼吸道合胞病毒感染后Treg/Th17及TH1/Th2失衡的调节机制研究

Respiratory syncytial virus (RSV) is the dominant cause of severe lower respiratory tract infection in children. So far, western medicine has no effective prophylaxis or treatment against RSV infection. Previous studies showed that the pathogenic mechanism of RSV is closely associated with the drift of Th subsets, especially with the unbalance of Treg/Th17 and Th1/Th2. Our long-term observation on clinical efficacy and former experimental studies have showed that the traditional Chinese prescription Qingfei oral liquid, which was developed independently, is very effective in RSV therapy. Furthermore, the therapeutic mechanism of Qingfei oral liquid has a close relationship with its influence on relative gene expressions of Th cells, as well as its regulation of immune function. In this project, effects of Qingfei oral liquid on the balance of Th subsets and its regulative function on the unbalance of Treg/Th17 and Th1/Th2 during RSV infection will be systematically studied in vivo. Changes of Treg/Th17 and Th1/Th2 will be detected by flow cytometry. Expressions of IL10, IL17, IL4 and IFN-γ will be detected by ELISA. The mechanism of Qingfei oral liquid in its anti-RSV effects and regulation of immune function will be studied from immunological and molecular cell biological perspectives, in order to reveal the molecular targets and delicate functional mechanism of Qingfei oral liquid in future.

呼吸道合胞病毒(RSV)是造成患儿严重下呼吸道感染的主导因素。目前，西医对RSV感染尚无有效预防和治疗方法。研究表明RSV致病机制与引起宿主Th细胞亚群漂移密切相关，尤其表现为Treg/Th17、Th1/Th2失衡。课题组通过长期临床疗效观察及前期实验研究，证实自主研制的开肺化痰解毒活血中药复方清肺口服液治疗RSV感染疾病疗效良好，并且能通过影响部分Th细胞相关基因的表达、调节机体免疫功能，以拮抗RSV感染。为系统观察清肺口服液对Th细胞亚群平衡的影响，探究其对RSV感染后Treg/Th17及Th1/Th2失衡的调节作用，本课题拟采用流式细胞术检测清肺口服液作用于动物模型前后Treg/Th17、Th1/Th2细胞数量的比例变化；用酶联免疫吸附法检测关键因子IL10、IL17、IL4及IFN-γ的表达水平变化，以从免疫学、分子细胞生物学角度探讨清肺口服液拮抗RSV、调节机体免疫功能的作用机理，为后续工作中确定药物作用的分子靶标及详细机制打下基础。

本课题旨在观察清肺口服液对Th细胞亚群平衡的影响，探究其对RSV感染后Treg/Th17及Th1/Th2失衡的调节作用。现已按预期目标完成实验，采用流式细胞术检测清肺口服液作用于动物模型前后Treg/Th17、Th1/Th2细胞数量的比例变化；用酶联免疫吸附法检测关键因子IL10、IL17、IL4及IFN-γ的表达水平变化。实验发现小鼠肺组织病理结果模型组与空白组相对比，模型组炎症渗出明显；小鼠模型组与各给药组小鼠相对比，各给药组均有不同程度 IL-4、IFN-γ及Th1/Th2的上调和IL-10、IL-17及Treg/Th17的下调。本实验从免疫学、分子细胞生物学角度揭示了清肺口服液拮抗RSV、调节机体免疫功能的作用机理，为后续工作中确定药物作用的分子靶标及详细机制打下基础。