Mahuang Decoction Antagonizes Acute Liver Failure via Modulating Tricarboxylic Acid Cycle and Amino Acids Metabolism

Abstract

Acute liver failure (ALF) is a serious clinical disorder with high fatality rates. Mahuang decoction (MHD), a well-known traditional Chinese medicine, has multiple pharmacological effects, such as anti-inflammation, anti-allergy, anti-asthma, and anti-hyperglycemia. In this study, we investigated the protective effect of MHD against ALF. In the lipopolysaccharide and D-galactosamine (LPS/D-GalN)-induced ALF mouse model, the elevated activities of the serum alanine and aspartate transaminases as well as the liver pathological damage were markedly alleviated by MHD. Subsequently, a metabolomics study based on the ultrahigh performance liquid chromatograph coupled with Q Exactive Orbitrap mass spectrometry was carried to clarify the therapeutic mechanisms of MHD against ALF. A total of 36 metabolites contributing to LPS/D-GalN-induced ALF were identified in the serum samples, among which the abnormalities of 27 metabolites were ameliorated by MHD. The analysis of metabolic pathways revealed that the therapeutic effects of MHD are likely due to the modulation of the metabolic disorders of tricarboxylic acid (TCA) cycle, retinol metabolism, tryptophan metabolism, arginine and proline metabolism, nicotinate and nicotinamide metabolism, phenylalanine metabolism, phenylalanine, tyrosine and tryptophan synthesis, as well as cysteine and methionine metabolism. This study demonstrated for the first time that MHD exerted an obvious protective effect against ALF mainly through the regulation of TCA cycle and amino acid metabolism, highlighting the importance of metabolomics to investigate the drug-targeted metabolic pathways.

Keywords: UPLC-Q-exactive-MS; acute liver failure; amino acids metabolism; mahuang decoction; metabolomics; tricarboxylic acid cycle.

FIGURE 1

Effects of MHD on serum ALT (A) and AST (B) levels in mice with acute liver failure induced by LPS/D-GalN. Values ​​are shown in Mean ± SEM. ***p < 0.001, ****p < 0.0001 vs model group. (C) Images of H&E stained liver cells (original magnification ×200).

FIGURE 2

PCA, PLS-DA and permutation test of serum samples. (A) PCA score scatter plots. (B) PLS-DA score scatter plots. (C) Permutation tests of PLS-DA.

FIGURE 3

OPLS-DA score plots, permutation test and S-plots of serum samples. (A) OPLS-DA score plots. (B) Permutation tests of OPLS-DA. (C) S-plots obtained from OPLS-DA model.

FIGURE 4

Heatmap of the protective activity of MHD for ALF. The degree of change is marked with different colors. Red and green indicate up-regulation and down-regulation respectively. Each row represents a metabolite. Each column represents an individual sample.

FIGURE 5

Changes in the relative intensity of target metabolites. Statistical significance was performed using univariate statistical analysis. 27 metabolites were significantly reversed by MHD. Values ​​are shown in Mean ± SEM. *p < 0.05 vs. model group.

FIGURE 6

Analysis of metabolic pathways (A) control vs model (B) MHD vs model. a: phenylalanine, tyrosine and tryptophan biosynthesis, b: phenylalanine metabolism, c: tryptophan metabolism, d: arginine and proline metabolism, e: nicotinate and nicotinamide metabolism, f: retinol metabolism, g: arginine biosynthesis, h: TCA cycle, i: tyrosine metabolism, j: alanine, aspartate and glutamate metabolism k: cysteine and methionine metabolism.

FIGURE 7

Schematic metabolic network of LPS/D-GalN-induced ALF and MHD modulation based on the KEGG database. Red: the levels of metabolites in model group were up-regulated compared to control group. Blue: the levels of metabolites in model group were down-regulated compared to control group. The italic metabolite names were significantly reversed by MHD treatment. Abbreviations: LysoPC, Lysophosphatidylcholine; GPC, Glycerophosphocholine.