Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Jun 24;19(6):e0305903.
doi: 10.1371/journal.pone.0305903. eCollection 2024.

Network pharmacology and experimental verification to decode the action of Qing Fei Hua Xian Decotion against pulmonary fibrosis

Hao-Liang Ke  1 , Rui-Jie Li  2 , Chao-Chao Yu  3 , Xiu-Ping Wang  1 , Chao-Yan Wu  1 , Ying-Wen Zhang  1
Affiliations

Network pharmacology and experimental verification to decode the action of Qing Fei Hua Xian Decotion against pulmonary fibrosis

Hao-Liang Ke et al. PLoS One. .

Abstract

Background: Pulmonary fibrosis (PF) is a common interstitial pneumonia disease, also occurred in post-COVID-19 survivors. The mechanism underlying the anti-PF effect of Qing Fei Hua Xian Decotion (QFHXD), a traditional Chinese medicine formula applied for treating PF in COVID-19 survivors, is unclear. This study aimed to uncover the mechanisms related to the anti-PF effect of QFHXD through analysis of network pharmacology and experimental verification.

Methods: The candidate chemical compounds of QFHXD and its putative targets for treating PF were achieved from public databases, thereby we established the corresponding "herb-compound-target" network of QFHXD. The protein-protein interaction network of potential targets was also constructed to screen the core targets. Furthermore, Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were used to predict targets, and pathways, then validated by in vivo experiments.

Results: A total of 188 active compounds in QFHXD and 50 target genes were identified from databases. The key therapeutic targets of QFHXD, such as PI3K/Akt, IL-6, TNF, IL-1β, STAT3, MMP-9, and TGF-β1 were identified by KEGG and GO analysis. Anti-PF effects of QFHXD (in a dose-dependent manner) and prednisone were confirmed by HE, Masson staining, and Sirius red staining as well as in vivo Micro-CT and immunohistochemical analysis in a rat model of bleomycin-induced PF. Besides, QFXHD remarkably inhibits the activity of PI3K/Akt/NF-κB and TGF-β1/Smad2/3.

Conclusions: QFXHD significantly attenuated bleomycin-induced PF via inhibiting inflammation and epithelial-mesenchymal transition. PI3K/Akt/NF-κB and TGF-β1/Smad2/3 pathways might be the potential therapeutic effects of QFHXD for treating PF.

PubMed Disclaimer

Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. The “herb-component-target” interaction network related to the effects of QFHXD on PF, based on the network pharmacology analysis.
In the network, the purple arrow represents the main herbs of QFHXD. The green and pink circles represent active components and targeted genes of QFHXD against PF, separately. HQ: Astragalus membranaceus, DG: Angelica sinensis, MH: Ephedra sinica Stapf, XR: Prunus armeniaca, BX: Pinellia ternata, GL: Trichosanthes kirilowii Maxim, TZS: Pseudostellaria heterophylla, TLZ: Lepidium apetalum Willd, CS: Paeonia lactiflora Pall, BL: Areca catechu L, ZS: Citrus aurantium L, ZBM: Fritillaria thunbergii Miq, SGL: Luffa cylindrica Roem, and GC: Radix Glycyrrhizae.
Fig 2
Fig 2. Venn diagram and PPI networks of the potential protein targets of the treatment with QFHXD.
(A) Venn diagram of the potential anti-PF targets. (B) PPI network of the protein targets. The circles represent the overlapped targets related to the effects of QFHXD on PF. (C) The top 20 core nodes in the PPI network are presented. The color depth and size of those core nodes are dependent on their degree values.
Fig 3
Fig 3. Enrichment analysis of the potential targets related to the effects of QFHXD on PF.
(A) The top 15 biological process (BP), cellular component (CC), and molecular function (MF) terms. (B) The top 20 KEGG pathways.
Fig 4
Fig 4. The “target-pathway” network.
The rose-red circle represents the potential targets related to QFHXD treatment. The green rhombus represents the main targeted pathways. The gray edges represent the interaction between the potential targets and their enriched pathways.
Fig 5
Fig 5. The representative micro-CT images and fibrotic score in each group.
(A) The representative micro-CT images in each group. (B) The quantity of micro-CT in rats by lung density parameter of Perc85. The data are presented as mean ± SD (n = 3 per group). #p<0.05 versus control group,*p<0.05 versus model group, ▲p<0.05 versus prednisone group, Δp<0.05 versus QFHXD-H group.
Fig 6
Fig 6. QFHXD and prednisone reduced the expression of collagen 1, collagen 3 and α-SMA in BLM-induced PF rats.
(A) Immunohistochemical staining of lung sections in each group. Magnification, ×400. Scale bar, 20μm. (B) Average IOD of collagen1 in each group. (C) Average IOD of collagen 3 in each group. (D) Average IOD of α-SMA in each group. The data are presented as mean ± SD (n = 3 per group). IOD, integral light density. #p<0.05 versus control group, *p<0.05 versus model group, ▲p<0.05 versus prednisone group, Δp<0.05 versus QFHXD-H group.
Fig 7
Fig 7. QFHXD and prednisone alleviated pathological changes in BLM-induced PF rats.
(A) Masson’s trichrome staining,Sirius red staining, and H&E staining of lung sections in each group. Magnification were ×200, ×400,and ×400, respectively. Scale bar were 50μm, 20μm, and 20μm, respectively. (B) Ratio of collagen in MT staining in each group. (C) Ratio of collagen in Sirius red staining in each group. (D) Ashcroft scale in each group. (E) Lung W/D weight ratio in each group. (F) MPO in lung in each group. The data are presented as mean ± SD (n = 3 per group). MT, Masson’s trichrome. #p<0.05 versus control group, *p<0.05 versus model group, ▲p<0.05 versus prednisone group, Δp<0.05 versus QFHXD-H group.
Fig 8
Fig 8. Effects of QFHXD and prednisone treatment on EMT in BLM-induced PF rats.
(A) α-SMA, fibronectin, E-cadherin, and vimentin protein expression in lung tissues in each group as measured by western-blot analysis. (B, C, D, E) The densitometry values for the proteins were normalized to those of β-actin. All data represent the mean ± SD of three independent experiments performed in triplicate. #p<0.05 versus control group, *p<0.05 versus model group, ▲p<0.05 versus prednisone group, Δp<0.05 versus QFHXD-H group.
Fig 9
Fig 9. Effects of QFHXD and prednisone treatment on MMP-9/TIMP1 balance and inflammation in BLM-induced PF rats.
(A, B, C) MMP-9, TIMP1, PI3K, p-PI3K, Akt, p-Akt, NF-κB p65, p-NF-κB p65, TNF-α, IL-6, and IL-1β protein expression in lung tissues in each group as measured by western-blot analysis. (D-N) The densitometry values for the proteins were normalized to those of β-actin. All data represent the mean ± SD of three independent experiments performed in triplicate. #p<0.05 versus control group, *p<0.05 versus model group, ▲p<0.05 versus prednisone group, Δp<0.05 versus QFHXD-H group.
Fig 10
Fig 10. Effects of QFHXD and prednisone treatment on TGF-β1/Smad2/3 signaling in BLM-induced PF rats.
(A, B, C) TGF-β1, p-Smad2, Smad2, p-Smad3, and Smad3 protein expression in lung tissues in each group as measured by western-blot analysis. (D-H)The densitometry values for the proteins were normalized to those of β-actin. All data represent the mean ± SD of three independent experiments performed in triplicate. #p<0.05 versus control group, *p<0.05 versus model group, ▲p<0.05 versus prednisone group, Δp<0.05 versus QFHXD-H group.
Fig 11
Fig 11. Schematic diagram showing how QFHXD alleviates BLM-induced pulmonary fibrosis by regulating PI3K/Akt/NF-κB and TGF-β1/Smad2/3 signaling pathways.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Raghu G. Epidemiology, survival, incidence and prevalence of idiopathic pulmonary fibrosis in the USA and Canada. Eur Respir J, 2017; 49. doi: 10.1183/13993003.02384-2016 - DOI - PubMed
    1. Gentile F, Aimo A, Forfori F, Catapano G, Clemente A, Cademartiri F, et al.. COVID-19 and risk of pulmonary fibrosis: the importance of planning ahead. Eur J Prev Cardiol, 2020; 27:1442��1446. doi: 10.1177/2047487320932695 - DOI - PMC - PubMed
    1. Raghu G, Rochwerg B, Zhang Y, Garcia CA, Azuma A, Behr J, et al.. An Official ATS/ERS/JRS/ALAT Clinical Practice Guideline: Treatment of Idiopathic Pulmonary Fibrosis. An Update of the 2011 Clinical Practice Guideline. Am J Respir Crit Care Med, 2015; 192:e3–19. doi: 10.1164/rccm.201506-1063ST - DOI - PubMed
    1. Maher TM, Corte TJ, Fischer A, Kreuter M, Lederer DJ, Molina-Molina M, et al.. Pirfenidone in patients with unclassifiable progressive fibrosing interstitial lung disease: a double-blind, randomised, placebo-controlled, phase 2 trial. Lancet Respir Med, 2020; 8:147–157. doi: 10.1016/S2213-2600(19)30341-8 - DOI - PubMed
    1. Wells AU, Flaherty KR, Brown KK, Inoue Y, Devaraj A, Richeldi L, et al.. Nintedanib in patients with progressive fibrosing interstitial lung diseases-subgroup analyses by interstitial lung disease diagnosis in the INBUILD trial: a randomised, double-blind, placebo-controlled, parallel-group trial. Lancet Respir Med, 2020; 8:453–460. doi: 10.1016/S2213-2600(20)30036-9 - DOI - PubMed

MeSH terms

Grants and funding

This work was supported by the Key Research & Development Project Funded by the Department of Science and Technology of Hubei, China (No. 2020BCB006). Ying-Wen Zhang received financial support which was provided by Department of Science and Technology of Hubei, China. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.