ulated and 33 downregulated substances (Figures 4A, B, G, H). There have been 151 distinct serum substances between group A and group C, with 113 upregulated and 28 downregulated substances (Figures 4C, D, I, J). Importantly, we discovered 174 distinct substances in the serum of group VA in comparison with group C, which includes 102 upregulated and 72 downregulated substances (Figures 4E, F, K, L).Ornithine Acts as a Therapeutic Target of VA Remedy in MM MiceTo narrow down the potential therapeutic targets, all the considerably diverse substances from every single comparison groups have been collected to plot Venn diagrams. Excluding the intersection, there had been 20 upregulated (Figure 5A and Table 1) and 32 downregulated (Figure 5B and Table 2) distinct metabolites in the serum of group VA. Subsequently, MetaboAnalyst 5.0 wasFrontiers in Oncology | frontiersin.orgNovember 2021 | Volume 11 | ErbB3/HER3 Inhibitor drug ArticleKe et al.Acupuncture and Bortezomib Advantage MMAEBFCGDHFIGURE two | Common chromatograms of TIC in serum samples. TIC of group C (A), V (B), A (C), and VA (D) in ESI+ mode. TIC of group C (E), V (F), A (G), and VA (H) in ESI- mode.Frontiers in Oncology | frontiersin.orgNovember 2021 | Volume 11 | ArticleKe et al.Acupuncture and Bortezomib Benefit MMABCDFIGURE 3 | PCA score plot based on the data of ESI+/- modes. (A) PCA score plot of all groups in ESI+ mode. (B) PCA score plot of all groups in ESI- mode. (C) 3D CDC Inhibitor Formulation scatter plot of all groups in ESI+ mode. (D) 3D scatter plot of all groups in ESI- mode.used to analyze the joint pathways of differential metabolites in ESI+ (Figure 5C) and ESI- (Figure 5D) modes, respectively. In Figure 5D, the principle pathway of arginine and proline metabolism was involved inside the serum of group VA with impact 0.20964 (-log (P)=4.6259). Ornithine and arginine had been important metabolites in arginine and proline metabolism pathway. Cluster evaluation and heatmap showed that both ornithine (median of m/z=133.1057307) and arginine (median of m/ z=347.2197103) were significantly decreased inside the serum of group VA compared with group C (Figure 5E). To a sizable degree, these information illustrated that VA therapy inhibited arginine and proline metabolism pathway, hence causing arginine and ornithine reduction. In addition, ornithine was also involved inside the regulation in the glutathione metabolicpathway (-log(P)=41.4122) in Figure 5D. These results suggested that ornithine may be a therapeutic target of VA therapy in MM.VA Treatment Decreases Ornithine Concentration in the Serum of MM MiceTo additional prove the above information, we performed targeted metabolomics to detect ornithine concentration within the serum of MM mice. The chromatogram revealed a characteristic peak of ornithine typical at 3.76 min immediately after injection (Figure 6A). In Figure 6B, based on the linear typical curve (r=0.99796), ornithine content within the serum samples of group VA (Average concentration of 7,333.33 ng/ml) was decreased by 73.36 compared with the group C (Typical concentration ofFrontiers in Oncology | frontiersin.orgNovember 2021 | Volume 11 | ArticleKe et al.Acupuncture and Bortezomib Benefit MMAGBHCIDJEKFLFIGURE 4 | OPLS-DA score plot depending on the data of ESI+/- modes and validations of OPLS-DA models by 200 permutation tests. In ESI+ mode: (A, B) group C vs group V, (C, D) group C vs group A, (E, F) group C vs group VA. In ESI- mode: (G, H) group C vs group V, (I, J) group C vs group A, (K, L) group C vs group VA.Frontiers in Oncology | frontiersin.orgNovember 2021 | Volume 11 |