# Four-dimensional data independent acquisition proteomics and metabolomics reveal mechanisms of hydrogen-rich water at Zusanli (ST36) point against triple-negative breast cancer in mice.
> 水素水の足三里（ST36）穴位注射による三陰性乳癌への作用：4次元データ非依存取得プロテオミクスおよびメタボロミクスによるメカニズム解析


## Abstract

This animal study examined the effects of hydrogen-rich water injected at the Zusanli (ST36) acupoint in mice bearing 4T1 breast cancer xenografts over a 21-day period. Four-dimensional data-independent acquisition (4D-DIA) proteomics identified differential protein expression in tumor tissue, highlighting Th1/Th2 cell differentiation pathways. Key proteins including IL-6 signal transducer, NFAT4, MAPK10, and MAPK11 were upregulated, while LAT, STAT1, and PKC-theta were downregulated. Serum metabolomics via LC-MS/MS revealed alterations in galactose, fructose, and mannose metabolism. Integrated multi-omics analysis pointed to the C-type lectin receptor signaling pathway as a central mechanism. Tumor growth was significantly suppressed and apoptosis of 4T1 cells was enhanced, findings corroborated by immunofluorescence, Western blotting, and RT-qPCR.

### Mechanism

Hydrogen-rich water injected at ST36 modulates Th1/Th2 differentiation and C-type lectin receptor signaling, upregulating MAPK10 and MAPK11 while downregulating LAT and STAT1, thereby promoting apoptosis in 4T1 breast cancer cells.

## Bibliographic

- **Authors**: Jingxuan L, Qian MO, Guowu L, Yejuan J, Aiying LI, Chunsheng J, et al.
- **Journal**: J Tradit Chin Med
- **Year**: 2025
- **PMID**: [41376217](https://pubmed.ncbi.nlm.nih.gov/41376217/)
- **DOI**: [10.19852/j.cnki.jtcm.20250929.002](https://doi.org/10.19852/j.cnki.jtcm.20250929.002)
- **PMC**: [PMC12711646](https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12711646/)
- **Study type**: animal study
- **Delivery route**: injection / infusion
- **Effect reported**: positive

## Delivery context

Intravenous hydrogen-saline infusion is a clinic-only route and is not viable for everyday self-administration. For routine hydrogen intake, inhalation is the most practical route, but inhalation carries explosion risk and concentration matters (empirical LFL of 10%; high-concentration 66% / 100% devices are not recommended).

## Safety notes

Intravenous hydrogen-saline infusion is a clinic-only route and is not viable for everyday self-administration. For routine hydrogen intake, inhalation is the most practical route, but inhalation carries explosion risk and concentration matters (empirical LFL of 10%; high-concentration 66% / 100% devices are not recommended).

See also:
- [Inhalation concentration and LFL / UFL](https://h2-papers.org/en/safety-notes/inhalation-concentration)
- [Consumer Affairs Agency accident cases](https://h2-papers.org/en/safety-notes/accident-cases)
- [Inhalation safety threshold lineage](https://h2-papers.org/en/safety-notes/lineage)

---

> **Cite as**: H2 Papers — PMID 41376217. https://h2-papers.org/en/papers/41376217
> **Source**: PubMed PMID [41376217](https://pubmed.ncbi.nlm.nih.gov/41376217/)