Exploring the Anti-Pulmonary Fibrosis Mechanism of Jingyin Granule by Network Pharmacology Strategy

Abstract

Pulmonary fibrosis (PF) is a clinically common disease caused by many factors, which will lead to lung function decline and even respiratory failure. Jingyin granule has been confirmed to have anti-inflammatory and antiviral effects by former studies, and has been recommended for combating H1N1 influenza A virus (H1N1) infection and Coronavirus disease 2019 (COVID-19) in China. At present, studies have shown that patients with severe COVID-19 infection developed lung fibrotic lesions. Although Jingyin granule can improve symptoms in COVID-19 patients, no study has yet reported whether it can attenuate the process of PF. Here, we explored the underlying mechanism of Jingyin granule against PF by network pharmacology combined with in vitro experimental validation. In the present study, the active ingredients as well as the corresponding action targets of Jingyin granule were firstly collected by TCMSP and literature data, and the disease target genes of PF were retrieved by disease database. Then, the common targets were subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, and then a PPI network and an ingredient-target network were constructed. Next, UPLC-MS was used to isolate and identify selected representative components in Jingyin granule. Finally, LPS was used to induce the A549 cell fibrosis model to verify the anti-PF effect of Jingyin granule in vitro. Our results indicated that STAT3, JUN, RELA, MAPK3, TNF, MAPK1, IL-6, and AKT1 were core targets of action and bound with good affinity to selected components, and Jingyin granule may alleviate PF progression by Janus kinase 2/signal transducers and activators of transcription (JAK2/STAT3), the mammalian nuclear factor-κB (NF-κB), the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt), tumor necrosis factor (TNF), and the extracellular signal-regulated kinases 1 and 2 (ERK1/2) signaling pathways. Overall, these results provide future therapeutic strategies into the mechanism study of Jingyin granule on PF.

Keywords: Jingyin granule; UPLC-MS; molecular mechanism; network pharmacology; pulmonary fibrosis; signaling pathway analysis.

FIGURE 1

Flow chart of network pharmacology research on the mechanism of Jingyin granule in anti-pulmonary fibrosis.

FIGURE 2

One hundred twenty-six components in Jingyin granule.

FIGURE 3

Venn diagram of common targets of pulmonary fibrosis and Jingyin granule.

FIGURE 4

Components–targets network of Jingyin granule against pulmonary fibrosis.

FIGURE 5

PPI network of potential pulmonary fibrosis targets acted by major components of Jingyin granule (A,B). The top 35 genes in the PPI network (C).

FIGURE 6

GO functional analysis of the shared targets of Jingyin granule and pulmonary fibrosis.

FIGURE 7

KEGG functional analysis of the shared targets of Jingyin granule and pulmonary fibrosis.

FIGURE 8

Target genes-pathways network of Jingyin granule.

FIGURE 9

(A) Molecular docking results of Jingyin granule components and selected targets. (B) The docking scores of Jingyin granule components and core targets.

FIGURE 10

Chromatograms of the seven compounds detected in Jingyin granule. (1: gallic acid, 2: chlorogenic acid, 3: rutin, 4: quercetin, 5: luteolin, 6: arctigenin, 7: kaempferol).

FIGURE 11

Effects of quercetin, arctigenin, gallic acid, chlorogenic acid, kaempferol, luteolin, and rutin on the protein expression of AKT, p-AKT, JAK2, p-JAK2, p44/42 MAPK (ERK1/2), p-p44/42 MAPK (ERK1/2), NF-κB, p-NF-κB, PI3K, p-PI3K, STAT3, p-STAT3, and TNF-α of LPS-induced A549 cells (A–N). *p < 0.05, **p < 0.01, compared with the LPS group.