Metabolomic Insights Into the Synergistic Effect of Biapenem in Combination With Xuebijing Injection Against Sepsis

Abstract

The drug combination of biapenem (BIPM) and xuebijing injection (XBJ) is commonly applied for the treatment of sepsis in China. However, the potential synergistic mechanism is still enigmatic. There have been no studies focused on the plasma metabolome alterations in sepsis after the intervention of this combination. In this work, an untargeted metabolomics approach was performed by liquid chromatography-mass spectrometry coupled with multivariate statistical analysis to provide new insights into the synergistic effect of BIPM in combination with XBJ. We characterized the metabolic phenotype of sepsis and described metabolic footprint changes in septic rats responding to XBJ and BIPM individually and in combination, in addition to histopathological and survival evaluation. A total of 91 potential biomarkers of sepsis were identified and 32 disturbed metabolic pathways were constructed. Among these biomarkers, 36 metabolites were reversely regulated by XBJ, mainly including glycerophospholipids, sphingolipids, free fatty acids (FFAs), bile acids and acylcarnitines; 42 metabolites were regulated by BIPM, mainly including amino acids, glycerophospholipids, and acylcarnitines; 72 metabolites were regulated after XBJ-BIPM combination treatment, including most of the 91 potential biomarkers. The results showed that the interaction between XBJ and BIPM indeed exhibited a synergistic effect by affecting some key endogenous metabolites, 15 metabolites of which could not be regulated when XBJ or BIPM was used alone. Compared with Model group, 13, 22, and 27 metabolic pathways were regulated by XBJ, BIPM, and XBJ-BIPM combination, respectively. It suggested that many more endogenous metabolites and metabolic pathways were significantly regulated after combination treatment compared with XBJ or BIPM monotherapy. Metabolisms of lipids, amino acids, acylcarnitines, and bile acids were common pathways involved in the synergistic action of XBJ and BIPM. This study was the first to employ metabolomics to elucidate the synergistic effect and decipher the underlying mechanisms of BIPM in combination with XBJ against sepsis. The results provide some support for clinical application of antibiotics in combination with traditional Chinese medicines and have important implications for the treatment of sepsis in clinic.

Keywords: biapenem; drug combination; metabolomics; synergistic effect; xuebijing injection.

Figure 1

PLS-DA and OPLS-DA score plots in positive and negative ion modes. PLS-DA score plots of different groups in positive (A) and negative (B) ion modes. OPLS-DA score plots for Model group vs. Sham group in positive (C) and negative (D) ion modes. Permutation tests obtained from Model group vs. Sham group in positive (E) and negative (F) ion modes. PLS-DA, partial least-squares discriminant analysis; OPLS-DA, orthogonal partial least-squares discriminant analysis.

Figure 2

(A) Venn diagram showing the number of regulated metabolites for different comparisons. (B) Pie charts showing the classification of regulated metabolites for different comparisons. XBJ, xuebijing injection; BIPM, biapenem; COM, combination of xuebijing injection and biapenem.

Figure 3

Heat map showing the level changing trend of 91 differential metabolites across the five groups. The colors from blue to red indicate higher levels of metabolites. XBJ, xuebijing injection; BIPM, biapenem; COM, combination of xuebijing injection and biapenem.

Figure 4

The average peak areas of each metabolite in each group for the 72 metabolites detected in positive (A) and negative (B) ion modes. ^#p < 0.05, ^##p < 0.01 versus the Sham group; ^*p < 0.05, ^**p < 0.01 versus the Model group. XBJ, xuebijing injection; BIPM, biapenem; COM, combination of xuebijing injection and biapenem.

Figure 5

Topology maps of the altered metabolic pathways analyzed by metaboanalyst between Model group vs. Sham group (A), XBJ group vs. Model group (B), BIPM group vs. Model group (C), COM group vs. Model group (D). XBJ, xuebijing injection; BIPM, biapenem; COM, combination of xuebijing injection and biapenem.

Figure 6

Disordered metabolic pathway networks in sepsis and the interventional effects of biapenem (BIPM) in combination with xuebijing injection (XBJ). The red dots represent increased metabolites in septic rats, the blue dots represent decreased metabolites in septic rats, and the gray dots represent undetected metabolites. The disturbed metabolic pathways in septic rats are marked in red (up-regulated) and blue (down-regulated). The metabolites regulated by the combination of XBJ and BIPM are marked with up and down arrows.