Integration of Metabolomics and Transcriptomics Reveals the Therapeutic Mechanism Underlying Paeoniflorin for the Treatment of Allergic Asthma

Abstract

Objectives: Asthma is a chronic airway inflammatory disease, which is characterized by airway remodeling, hyperreactivity and shortness of breath. Paeoniflorin is one of the major active ingredients in Chinese peony, which exerts anti-inflammatory and immune-regulatory effects in multiple diseases. However, it remains unclear whether paeoniflorin treatment can suppress allergic asthma. Methods: In this study, we evaluated the effect of paeoniflorin on lung function and airway inflammation in asthmatic mice. These asthmatic Balb/c mice were first sensitized and constructed through ovalbumin (OVA) motivation. Subsequently, we determined the mechanism of action of paeoniflorin in treating allergic asthma through integrated transcriptomic and metabolomic data sets. Results: Our results demonstrated that many genes and metabolites were regulated in the paeoniflorin-treated mice. Moreover, the potential target proteins of paeoniflorin played important roles in fatty acid metabolism, inflammatory response, oxidative stress and local adhesion. Conclusion: Paeoniflorin has a beneficial effect on asthma, which may be achieved through regulating fatty acid metabolism, inflammatory response and the adhesion pathway at system level.

Keywords: allergic asthma; fatty acid metabolism; metabolomics; paeoniflorin; transcriptomics.

FIGURE 1

Paeoniflorin reduce inflammation and airway hyperresponsiveness in allergic asthma mice. The staining of BALF (A), lung tissue pathological changes and inflammation inflammatory infiltration (B), Leukocytes, EOS, and neutrophils level in BALF (C), and Penh level (D). The data were expressed as the mean ± SD. ^∗Means control group vs model group (^∗∗∗p < 0.001); ^#Means model group vs paeoniflorin group (^##p < 0.01 and ^###p < 0.001).

FIGURE 2

Paeoniflorin reduces chemokines and cytokines in allergic asthma mice. The level of exotaxin in serumand lung tissue (A), the level of MIP-1α in serum and lung tissue (B), the level of IL-17 in serum and lung tissue (C), and the level of IL-4 in serumand lung tissue (D). The data were expressed as the mean ± SD. ^∗Means control group vs model group (^∗p < 0.05, ^∗∗p < 0.01, and ^∗∗∗p < 0.001); ^#Means model group vs paeoniflorin group (^#p < 0.05, ^##p < 0.01, and ^###p < 0.001).

FIGURE 3

Metabonomic analysis results of paeoniflorin-treated OVA-induced allergic asthma mice. Score plots from the PLS-DA model (A) and heatmap (B) classifying the control group (Green), model group (Red), paeoniflorin group (Blue) with mice lung samples by UPLC-Q-TOF/MS. Metabolites set enrichment results was shown in C and D.

FIGURE 4

Transcriptomics analysis of OVA on normal mouse. (A) GO enrichment analysis of DEGs between Model and Control in BP (Biological process), MF (Molecular function) and Cellular Component (CC). (B–D) GSEA analysis of DEGs between Model and Control: (B) mmu05310: Asthma; (C) mmu04371: Apelin signaling pathway; (D) mmu04022: cGMP-PKG signaling pathway.

FIGURE 5

Transcriptomics analysis of paeoniflorin on allergic asthma model mouse. (A) GO enrichment analysis of DEGs between Model and Paeoniflorin in BP (Biological process), MF (Molecular function) and Cellular Component (CC). (B–D) GSEA analysis of DEGs between Model and Control: (B) mmu05152: Tuberculosis; (C) mmu04510: Focal adhesion; (D) mmu04060: Cytokine-cytokine receptor interaction.

FIGURE 6

The enrichment analysis of candidate genes of paeoniflorin on allergic asthma model mouse. (A) Heatmap classifying the candidate genes of paeoniflorin on allergic asthma model mouse. (B) mRNA expression level of Ttl, Dusp, Map3k7 and Asah1f was detected by real-time PCR assay. (C) The genes were mainly related to such biological functions: fatty acid metabolism, inflammation response, cytokinesis, chromosome and adhesion plaque. The data were expressed as the mean ± SD. ^∗Means control group vs model group (^∗p < 0.05, ^∗∗p < 0.01, and ^∗∗∗p < 0.001); ^#Means model group vs paeoniflorin group (^#p < 0.05, ^##p < 0.01, and ^###p < 0.001).

FIGURE 7

Integrated analysis the mechanism of paeoniflorin-treated OVA-induced allergic asthma mouse from metabonomics and transcriptomics data. Compound reaction networks of the metabolites, genes were visualized using Metscape: metabolites (hexagons), genes (circles), metabolic enzymes (squares) and chemical reactions (grey rhombus) are presented as nodes and relationship are presented as edges. Inputted genes are shown in blue, inputted metabolites are shown in red. The metabolite-gene associated network was mainly related to fatty acid metabolism.