Beijing University of Chinese Medicine, Beijing, PR China; Union Stem Cell & Gene Engineering Co., Ltd, Tianjin, PR China.
Beijing University of Chinese Medicine, Beijing, PR China.
J Ethnopharmacol. 2024 Sep 15;331:118159. doi: 10.1016/j.jep.2024.118159. Epub 2024 Apr 25.
Tanreqing injection (TRQ) is widely used, traditional Chinese medicine (TCM) injection used in China to treat respiratory infections. Modern pharmacological studies have confirmed that TRQ can protect against influenza viruses. However, the mechanism by which TRQ inhibits influenza viruses remains unclear.
To explore the therapeutic effects and possible mechanisms of TRQ inhibition by the influenza virus.
Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF MS) was used to determine the chemical composition of TRQ. Isobaric tags for relative and absolute quantification (iTRAQ) were used to define differential proteins related to TRQ inhibition of viruses. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed for functional annotation. For experimental validation, we established an in vitro model of the influenza virus infection by infecting A549 cells with the virus. The detection of the signaling pathway was carried out through qPCR, western blotting,and immunofluorescence.
Fifty one components were identified using UPLC/Q-TOF MS. We confirmed the inhibitory effect of TRQ on influenza virus replication in vitro. Ninety nine differentially expressed proteins related to the inhibitory effect of TRQ were identified using iTRAQ. KEGG functional enrichment analysis showed that the TRQ may inhibit influenza virus replication by affecting autophagy. Through network analysis, 29 targets were selected as major targets, and three key targets, HSPA5, PARP1, and GAPDH, may be the TRQ targets affecting autophagy. In vitro experiments showed that TRQ inhibits influenza virus replication by interfering with the expression and localization of STX17 and VAMP8 proteins, thereby promoting the fusion of autophagosomes with lysosomes.
TRQ inhibits influenza virus replication by promoting the fusion of autophagosomes with lysosomes. We additionally established potential gene and protein targets which are affected by TRQ. Therefore, our findings provide new therapeutic targets and a foundation further studies on influenza treatment with TRQ.
痰热清注射液（TRQ）是一种广泛应用于中国治疗呼吸道感染的中药注射液。现代药理学研究已经证实，TRQ 可以预防流感病毒。然而，TRQ 抑制流感病毒的机制尚不清楚。
探讨 TRQ 抑制流感病毒的治疗效果和可能机制。
采用超高效液相色谱-四级杆飞行时间质谱联用（UPLC/Q-TOF MS）测定 TRQ 的化学成分。采用同位素相对和绝对定量标记（iTRAQ）技术鉴定与 TRQ 抑制病毒相关的差异蛋白。对京都基因与基因组百科全书（KEGG）通路进行功能注释。为了进行实验验证，我们通过感染 A549 细胞建立了流感病毒体外感染模型。通过 qPCR、western blot 和免疫荧光检测信号通路。
采用 UPLC/Q-TOF MS 鉴定出 51 个成分。我们证实了 TRQ 在体外抑制流感病毒复制的作用。通过 iTRAQ 鉴定出 99 个与 TRQ 抑制作用相关的差异表达蛋白。KEGG 功能富集分析表明，TRQ 可能通过影响自噬来抑制流感病毒的复制。通过网络分析，选择了 29 个主要靶点，其中 3 个关键靶点 HSPA5、PARP1 和 GAPDH，可能是 TRQ 影响自噬的靶点。体外实验表明，TRQ 通过干扰 STX17 和 VAMP8 蛋白的表达和定位，抑制流感病毒的复制，从而促进自噬体与溶酶体的融合。
TRQ 通过促进自噬体与溶酶体的融合来抑制流感病毒的复制。我们还建立了可能受 TRQ 影响的潜在基因和蛋白质靶点。因此，我们的研究结果为进一步研究 TRQ 治疗流感提供了新的治疗靶点和基础。
