Dissection of Biological Property of Chinese Acupuncture Point Zusanli Based on Long-Term Treatment via Modulating Multiple Metabolic Pathways

Abstract

Acupuncture has a history of over 3000 years and is a traditional Chinese medical therapy that uses hair-thin metal needles to puncture the skin at specific points on the body to promote wellbeing, while its molecular mechanism and ideal biological pathways are still not clear. High-throughput metabolomics is the global assessment of endogenous metabolites within a biologic system and can potentially provide a more accurate snap shot of the actual physiological state. We hypothesize that acupuncture-treated human would produce unique characterization of metabolic phenotypes. In this study, UPLC/ESI-HDMS coupled with pattern recognition methods and system analysis were carried out to investigate the mechanism and metabolite biomarkers for acupuncture treatment at "Zusanli" acupoint (ST-36) as a case study. The top 5 canonical pathways including alpha-linolenic acid metabolism, d-glutamine and d-glutamate metabolism, citrate cycle, alanine, aspartate, and glutamate metabolism, and vitamin B6 metabolism pathways were acutely perturbed, and 53 differential metabolites were identified by chemical profiling and may be useful to clarify the physiological basis and mechanism of ST-36. More importantly, network construction has led to the integration of metabolites associated with the multiple perturbation pathways. Urine metabolic profiling might be a promising method to investigate the molecular mechanism of acupuncture.

Figure 1

Acupuncture (a) and acupoint map of human meridians (b). c: “Zusanli” acupoint (ST-36).

Figure 2

PCA model results for control and model group in positive mode (a). Trajectory analysis of PCA score plots (3D) for the serum samples in positive mode (b). PCA model results for 0 day and 14 days in positive mode (c). VIP plot of OPLS-DA of samples in positive mode (d).

Figure 3

PCA model results for control and model group in negative mode (a). Trajectory analysis of PCA score plots (3D) for the serum samples in negative mode (b). PCA model results for 0 day and 14 days in negative mode (c). VIPplot of OPLS-DA of samples in negative mode (d).

Figure 4

Mass fragment information of alpha-N-phenylacetyl-L-glutamine in negative mode (a) and 9-decenoylcarnitine in positive mode (b).

Figure 5

Construction of the arachidonic acid 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: alpha-linolenic acid metabolism; 2: D-glutamine and D-glutamate metabolism; 3: citrate cycle (TCA cycle); 4: alanine, aspartate, and glutamate metabolism; 5: vitamin B6 metabolism.