基于网络药理学研究清瘟解热合剂治疗COVID-19的多靶协同作用机制

清瘟解热合剂在新冠疫情中具有较好的临床疗效。基于前期研究基础，我们推测清瘟解热合剂可能通过干扰SARS-CoV-2中S蛋白与ACE2结合抑制病毒感染，调控CCL3/CCR5信号通路，抑制巨噬细胞迁移及活化发挥作用。项目拟通过①构建SARS-CoV-2假病毒以及过表达ACE2的细胞系，研究含药血清对SARS-CoV-2与靶细胞受体结合的抑制作用；②建立病毒感染小鼠急性肺损伤模型，结合流式、液相芯片等技术，分析清瘟解热合剂对小鼠急性损伤、巨噬细胞活化及CCL3/CCR5信号通路的影响；③构建巨噬细胞与肺泡上皮细胞共培养体系，结合RNAi等分子生物学方法，探讨沉默或过表达CCR5后，清瘟解热合剂对体外巨噬细胞活化的影响；从抗病毒及抗炎作用两方面，探讨清瘟解热合剂治疗COVID-19的多靶作用机制，阐明清瘟解热合剂治疗COVID-19的可能靶点，为中医治疗冠状病毒感染性疾病提供研究基础。

Qingwen Jiere mixture presents good clinical efficacy during the outbreak of COVID-19. Based on the previous research, we speculated that Qingwen Jiere mixture might inhibit viral infection by interfering with the binding of S protein to ACE2 in SARS-CoV-2, regulate the CCL3/CCR5 signaling pathway, inhibit macrophage migration and activation and alleviate pulmonary inflammatory response, thus playing a role in coronavirus disease 2019 (COVID-19) treatment. This project intends to ① investigate the inhibitory effect of drug-containing serum on the binding of SARS-CoV-2 to the receptor of target cells by constructing SARS-CoV-2 pseudovirus and cell line with ACE2 overexpression, ② analyze the effects of Qingwen Jiere mixture on acute injury, macrophage activation and the CCL3/CCR5 signaling pathway in mice by establishing an acute lung injury model in virus-infected mice using flow cytometry and liquid-phase chip, and ③ explore the effect of Qingwen Jiere mixture on macrophage activation in vitro after silencing or overexpression of CCR5 by constructing a co-culture system of macrophages and alveolar epithelial cells using RNAi and other molecular biological methods. Moreover, the multi-target synergistic mechanism of Qingwen Jiere mixture in the treatment of COVID-19 is explored from antiviral and anti-inflammatory effects, and the potential targets of Qingwen Jiere mixture in treating COVID-19 are clarified, which provides a research basis for COVID-19 treatment by traditional Chinese medicine.