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STUDY 03 / METABOLOMICS

Short-term changes in the blood metabolome

A small crossover study exploring blood metabolites 10 and 30 minutes after Kanshoho. Within-condition changes and between-condition differences are reported separately.

Conference presentation · 2025
Analysed
9 adults with low back pain
Measured
580 metabolites
Sampling
Before · 10 min · 30 min

Design

Randomised, single-blind crossover trial; nine adults with low back pain were analysed. Kanshoho and a control procedure were given 2 weeks apart, and 580 blood metabolites were measured with HMT Dual Scan. Trial registration: UMIN000049180.

Crossover design

Kanshoho
2 weeks
Control
Control
2 weeks
Kanshoho

Each participant received both conditions in the allocated order.

BeforeBlood sample
10 min afterBlood sample
30 min afterBlood sample
Summary of the design and sampling times. Source: slides, 34th Annual Meeting of the Japanese Society of Pathophysiology, 2025 (Japanese) .

What was reported

The presentation highlights 21 metabolites that changed from baseline within the Kanshoho condition. “First significant” in the list below means the first time point at which the within-condition comparison with baseline reached P<0.05. Comparison with the control procedure is reported alongside.

ExampleWithin-condition changeBetween-condition P value
Retinol-2Increased at 10 and 30 min10 min: 0.4597 / 30 min: 0.0987
Acylcarnitine (13:1)-3Increased at 10 and 30 min10 min: 0.8321 / 30 min: 0.3935

For both examples the between-condition comparison gave P≥0.05. The within-condition changes are therefore a starting list of candidates; effect sizes, confidence intervals and time courses against the control are the next step.

All 21 metabolites

Nine metabolites increased and twelve decreased. Names and suffix numbers follow the presentation.

Metabolite (as named in the presentation)DirectionFirst significant
3β-Hydroxy-5-cholestenoic acid-3Increase30 min
5β-Tetrahydrocortisol-1Decrease30 min
Acylcarnitine (13:1)-3Increase10 min
Acylcarnitine (17:0)-2Increase30 min
Auraptene-1Increase30 min
BiliverdinDecrease30 min
cis-Aconitic acidDecrease10 min
Dehydroisoandrosterone 3-sulfate-2Increase30 min
Deoxycholic acidDecrease30 min
Gluconic acidDecrease10 min
Glycochenodeoxycholic acidDecrease10 min
HomoarginineIncrease30 min
N1-AcetylspermidineDecrease30 min
N2-PhenylacetylglutamineDecrease30 min
Pyrocatechol sulfateDecrease10 min
Retinol-2Increase10 min
Ricinoleic acid-3Increase30 min
SerIncrease30 min
Taurolithocholic acid 3-sulfateDecrease10 min
Trilaurin-4Decrease30 min
β-Estradiol-117/α-EstradiolDecrease30 min

Source: presentation slides, p. 4 (Japanese)

From metabolites to testable questions

Research direction

Direction of interestObserved candidatesQuestion to test
Energy metabolism and fatigueAcylcarnitine (13:1)-3 and (17:0)-2 increased; cis-aconitic acid decreasedHow do blood levels relate to substrate use, metabolic flux and fatigue when all are measured together?
Inflammation and redox stateRetinol-2 and auraptene-1 increased; biliverdin decreasedDo inflammatory markers, redox indices and symptoms move on the same time axis?
Endocrine, stress and pain5β-Tetrahydrocortisol-1 and estradiol decreased; dehydroisoandrosterone 3-sulfate-2 increasedWhich step — production, metabolism or excretion — is involved, and does it track pain or stress?
Bile acids and the gutDeoxycholic acid, glycochenodeoxycholic acid and others decreasedDoes the pattern replicate when meals, sampling time and gut-derived influences are standardised?
Polyamines and amino acidsN1-Acetylspermidine and N2-phenylacetylglutamine decreased; Ser increasedCan related metabolites, measured together, be linked to the local tissue response or to symptoms?

Pathway analysis

Pathway (as named in the presentation)Reported P valueReading
Steroid hormone biosynthesis0.0104Candidate steroid-related pathway
Retinol metabolism0.0923P≥0.05; impact 0.263. To be explored with related markers
Glyoxylate metabolism0.012Pathway name as given; correspondence with human metabolism to be examined

Enrichment and impact scores place the changed metabolites on pathways and help choose what to verify next. The P values are those shown in the presentation (MetaboAnalyst, slide 8).

Scope and next steps

Nine participants and 580 metabolites make this a hypothesis-generating screen.

Research direction

A proposed collaborative study would quantify the candidates against reference standards in an independent sample, with meals, activity and sampling time standardised, and to report between-condition differences with effect sizes and confidence intervals, including order and carry-over effects. Recording lipid use, inflammation and pain on the same time axis would help investigate which physiological functions the blood changes correspond to.

Discuss a metabolomics or blood-marker study

Ethics and funding

Approved by the ethics review board of Ueno Asagao Clinic; written informed consent was obtained. The study was funded by the Japan Health Organization. Safety recording methods and totals will be added to this page when the full paper is reported.

A related manuscript is titled “Comprehensive analysis of changes in blood metabolites after muscle relaxation treatment Kanshoho: a randomized, single-blind, crossover study”.

Sources

  1. Sakato T, Nakajima A. Rapid changes in blood metabolome induced by the low-pressure muscle relaxation technique “Kanshoho”: a randomized single-blind crossover trial. 34th Annual Meeting of the Japanese Society of Pathophysiology, 2025 (presentation slides, PDF, in Japanese)

Sections marked “Research direction” describe hypotheses and proposed studies for discussion with collaborators. Observations are presented with their participants, methods and sources.

Last updated: 21 September 2026

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