Deciphering the biological effects of acupuncture treatment modulating multiple metabolism pathways

Abstract

Acupuncture is an alternative therapy that is widely used to treat various diseases. However, detailed biological interpretation of the acupuncture stimulations is limited. We here used metabolomics and proteomics technology, thereby identifying the serum small molecular metabolites into the effect and mechanism pathways of standardized acupuncture treatments at 'Zusanli' acupoint which was the most often used acupoint in previous reports. Comprehensive overview of serum metabolic profiles during acupuncture stimulation was investigated. Thirty-four differential metabolites were identified in serum metabolome and associated with ten metabolism pathways. Importantly, we have found that high impact glycerophospholipid metabolism, fatty acid metabolism, ether lipid metabolism were acutely perturbed by acupuncture stimulation. As such, these alterations may be useful to clarify the biological mechanism of acupuncture stimulation. A series of differentially expressed proteins were identified and such effects of acupuncture stimulation were found to play a role in transport, enzymatic activity, signaling pathway or receptor interaction. Pathway analysis further revealed that most of these proteins were found to play a pivotal role in the regulation of multiple metabolism pathways. It demonstrated that the metabolomics coupled with proteomics as a powerful approach for potential applications in understanding the biological effects of acupuncture stimulation.

Figure 1. Trajectory analysis of PCA Score plots for the acupuncture treatment.

(△: the 1st day; □: the 3rd day; *: the 5th day; ●: the 7th day; +: the 9th day; ○: the 11th day; ▽: the 13th day; ▲: the 14th day).

Figure 2. Metabolic profling characterization and pattern recognition analysis.

(A) UPLC-MS BPI serum chromatograms of acupuncture-treated human in positive mode (a, 0 day; b, 14 day) and negative mode (c, 0 day; d, 14 day); (B) PCA score plot for control and acupuncture-treated group in positive mode; (C) Trajectory analysis of PCA score plots (3-D) for the serum samples in negative mode.

Figure 3. The screening of the metabolite markers.

(A) PLS loading plot results for 0 day and 14 day in positive mode; (B) VIP-plot of OPLS-DA of samples in positive mode; (C) PLS loading plot results for 0 day and 14 day in negative mode; (D) VIP-plot of OPLS-DA of samples in negative mode.

Figure 4. Identification of the important differential metabolites.

(A) Mass fragment information of LysoPE(P-16:0/0:0) in negative mode. (B) Construction of the metabolism pathways in acupuncture-treated human. The map was generated using the reference map by KEGG (http://www.genome.jp/kegg/). The green boxes: enzymatic activities with putative cases of analogy in acupuncture-treated human. 1, glycerophospholipid metabolism; 2, ether lipid metabolism; 3, fatty acid metabolism; 4, glycerolipid metabolism; 5, porphyrin metabolism; 6, sphingolipid metabolism; 7, primary bile acid biosynthesis; 8, fatty acid elongation in mitochondria; 9, fatty acid biosynthesis; 10, tryptophan metabolism.