Identification and chemical profiling of anti-alcoholic liver disease biomarkers of ginseng Huang jiu using UPLC-Q-Orbitrap-HRMS and network pharmacology-based analyses

Abstract

This study investigated the mechanism of characteristic non-volatile organic compounds (NVOCs) from ginseng Huang jiu (GH) in the treatment of alcoholic liver disease through UPLC-Q-Orbitrap-HRMS and network pharmacological analyses. Changes in NVOC contents in ginseng Huang jiu and ginseng-soaked wine fermented by different processing technologies were analyzed through liquid chromatography-mass spectrometry (LC-MS). A total of 96 ginsenosides were identified in ginseng Huang jiu throughout the fermentation process, which included 37 protopanaxadiol-type ginsenosides, 47 protopanaxatriol-type ginsenosides, and 4 oleanolic acid-type ginsenosides. Orthogonal partial least squares-discriminant analysis (OPLS-DA) revealed that 20(R)-Rg2, Gypenoside XVII, 20(S)-Rf3, CK, Rg5, Rh2, and other rare ginsenosides in ginseng Huang jiu could be the potential index for determining ginseng Huang jiu. In addition, ginseng Huang jiu could improve alcoholic liver disease by regulating the GSTP1, HRAS, AKR1B1, GSTA1, Androgen receptor (AR), GSR, and LDHB genes through bioinformatics analysis. This study provides new insights into improving the industrial production of ginseng Huang jiu and treating alcoholic liver disease with medicinal and food products.

Keywords: UPLC-Q-Orbitrap-HRMS; ginseng Huang jiu; ingredient analysis; network pharmacology; orthogonal partial least squares-discriminant analysis.

Figure 1

Representative UPLC-Q-Orbitrap-HRMS total ion chromatograph of ginseng Huang jiu and ginseng-soaked Wine.

Figure 2

Tandem mass spectrum of ginsenosides in the negative ion mode: Gypenoside XVII.

Figure 3

Possible transformation pathways of PPD-type ginsenosides under the action of GH.

Figure 4

Tandem mass spectrum of ginsenosides in the negative ion mode: 20-O-Glc-Rf.

Figure 5

Possible transformation pathways of PPT-type ginsenosides under the action of GH.

Figure 6

Tandem mass spectrum of ginsenosides in the negative ion mode: Ro.

Figure 7

Possible transformation pathways of OLE-type ginsenosides under the action of GH.

Figure 8

Heat map and HCA clustering results of 67 non-volatile compounds with a significant difference (p < 0.05) and aroma descriptors in ginseng wine samples fermented with different processes.

Figure 9

Multivariate statistical analysis of UPLC-Q-Orbitrap-HRMS metabolic profiling data. Principal component analysis (PCA) score plot (A), pair-wise orthogonal projections to latent structure discriminant analysis (OPLS-DA) score plot (B), and OPLS-DA/permutation test/S-plot (C) derived from the UPLC-Q-Orbitrap-HRMS spectra of both groups of ginseng Huang jiu (GH1–GH6, red triangles) and ginseng-soaked wine (GSW1–GSW6, green square) in negative ion mode. Statistical validation of the OPLS-DA model by permutation testing (D).

Figure 10

(A) A volcano plot was constructed using the fold change values and P-adjust. Red and blue dots indicate upregulated and downregulated genes, respectively. (B) Heatmap of the differential gene expression. Different colors represent the trend of gene expression in different tissues. The top 50 upregulated and top 50 downregulated genes are shown in (C): intersected mRNAs from GeneCards databases, ginseng Huang jiu, and ALD.

Figure 11

(A,B) GO and KEGG enrichment analyses for DEGs, terms with p and q < 0.05 were believed to be enriched significantly.

Figure 12

Identification of hub genes from common DEGs. (A) The STRING database constructed a PPI network of the common DEGs. (B) Seven hub genes were identified by the Cytohubba tool in Cytoscape.