Integrating pharmacokinetics and network pharmacology to decipher the efficacy of Fufang Qinlan oral liquid on post infectious cough

Abstract

Background and aim: Fufang Qinlan oral liquid (QLOL) is a traditional Chinese medicine formula used to treat fevers, sore throats and cough. This study was to evaluate QLOL's therapeutic effects on post infectious cough (PIC) and to investigate Q-markers and action mechanisms by integrating pharmacokinetics and network pharmacology.

Materials and methods: PIC rats were measured for cough frequency, organ index, pathological section of lung and inflammatory factors to evaluate the effects of QLOL. Pharmacokinetic analyses were performed using HPLC-QQQ-MS/MS to explore the possible Q-markers of QLOL and the changes of them in vivo. Network pharmacology was used to obtain potential important targets and action mechanisms of treating PIC. The possible hub genes were evaluated using QPCR.

Results: The symptoms of cough and lung injury were significantly alleviated and IL-6 and IL-1β were significantly decreased after treatment of QLOL. 6 compounds were considered as possible Q-makers and their pharmacokinetic parameters were analyzed. The compound-target network was constructed to identify 53 important targets. Among them, HSPA8, HSP90AA1, HSP90AB1, YWHAZ, EGFR, ESR1 and EP300 were selected as the core targets because of high degree value and direct connection with inflammation or microbial infection. All 6 compounds had potently binding activity to core targets in molecular docking. The QPCR results showed the up-regulation of core targets expression in QLOL group compared with PIC rats, which validated the effects of QLOL and the accuracy of Q-markers selection.

Conclusion: QLOL alleviated PIC symptoms through various targets and mechanisms. The potential Q-markers were baicalein, baicalin, oroxylin A-7-O-glucuronideloside, wogonoside, oroxylin A and forsythoside E.

Taxonomy classification by evise: the experimental approach.

Keywords: Fufang qinlan oral liquid; Pharmacokinetics; Post infectious cough; Potential Q-markers; Traditional Chinese medicine formula.

Graphical abstract

Fig. 1

The workflow of this study. The figure created with Biorender.com.

Fig. 2

QLOL alleviated PIC in rats. The lung function was measured by calculating cough time (A) and the organ index of lung (B).

Fig. 3

Representative H&E stained sections of lung of rats from the groups of control, PIC, QLOL and MSCT. The blue arrows indicated thickening of the alveolar septa. Black arrows indicated secretions or bleeding in the airway cavity. Yellow arrows indicated inflammatory cell infiltration. Green arrows indicated alveolar hemorrhage.

Fig. 4

Effect of QLOL on inflammatory cytokines. The expression levels of IL-1β (A), IL-6 (B) in the lung tissue were determined by ELISA. n = 7, x ± s. *P < 0.05 vs model group, **P < 0.01, ***P < 0.001 vs model group.

Fig. 5

Mean concentration-time curves of baicalin (A), forsythoside E (B), oroxylin A-7-O-glucuronide (C), wogonoside (D), oroxylin A (E) and baicalein (F) in rat blood after oral administration of QLOL.

Fig. 6

Content of 8 ingredients in QLOL (A) and the AUC values of the 8 components (B).

Fig. 7

Compound-target network of 6 ingredients in QLOL for cough. Purple nodes represented the ingredients, red nodes represented important targets. Node color depth of targets was proportional to the number of degrees.

Fig. 8

GO analysis (A) and KEGG pathway (B) of important genes of QLOL and PIC. In C, color means enriched type with green, orange and purple present represent biological processes (BP), cellular components (CC) and molecular functions (MF), respectively.

Fig. 9

Heat map of molecular docking results (A). The binding poses of HSPA8 complexed with baicalin, YWHAZ complexed with wogonoside, baicalin, oroxylin A-7-O-glucuronide and montelukast, EP300 complexed with montelukast (B).

Fig. 10

The effect of QLOL on the expression of key targets related to inflammation after PIC. A–G: Quantitative analysis of EP300, ESR1, HSPA8, YWHAZ, HSP90AA1, HSP90AB1, EGFR, ESR1, EP300. n = 6, x ± s. *P < 0.05, **P < 0.01, ***P < 0.001 vs model group.

Fig. 11

Pathway-effect relationships for QLOL. The icons on the left corresponded to the pathways enriched with KEGG. The icon on the right represented the main effects of these pathways.

Fig. 12

Mapping diagram of 6 ingredients, important targets and main pathways. The dots of different colors represented 6 ingredients. The dots under different targets corresponded to the compounds and targets connection in the compound-target network.