基于“TF—miRNA—NP”途径在宿主对抗流感病毒中的作用探讨金柴抗病毒胶囊的作用机制

金柴抗病毒胶囊为本课题组自主研发的中药6.1类新药，具有透邪解毒的功效，主治病毒性上呼吸道感染。前期研究发现，金柴抗病毒胶囊对治疗H1N1流感病毒感染有效，可以抑制流感病毒NP基因（流感病毒复制必需基因）的表达，从而阻碍病毒复制，同时可以上调宿主miRNA-431-5p的表达。申请人已经证实，在生物体外miRNA-431-5p可以下调流感病毒NP基因表达，并通过生物信息学预测转录因子（TF）NF-AT1、AML1、HIF-1α可以调控miRNA-431-5p表达。上述结果提示:宿主抗流感病毒可能通过“TF-miRNA-431-5p-NP”途径，金柴抗病毒胶囊通过调控此途径发挥药理作用。申请人将通过染色质免疫共沉淀（CHIP）、基因沉默、qPCR、Wetern Blot、细胞转染等技术验证这一假说。

Jinchai antiviral capsules for the research group independent research and development of Chinese medicine 6.1 new drugs, with the effect of clearing evil, detoxification, attending viral upper respiratory tract infection. Previous studies have found that Jinchai Virus Capsule is effective for the treatment of H1N1 influenza virus infection and can inhibit the expression of the influenza virus NP gene (indispensable gene for influenza virus replication), thereby hindering virus replication and at the same time it can upregulate the expression of host miRNA-431-5p. Applicants have confirmed that miRNA-431-5p can downregulate the expression of influenza virus NP gene in vitro and bioinformatics predict transcription factor NF-AT1, AML1, HIF-1α regulate miRNA-431-5p expression. The above results suggest that the host anti-influenza virus may pass the “transcription factor-miRNA-431-5p-NP” pathway, and Jinchai antiviral capsules exert pharmacological effects through regulating this pathway. Applicants will validate this hypothesis by techniques such as Chromatin Immunoprecipitation (CHIP), Gene silencing, qPCR, Wetern Blot, and Cell Transfection.