Molecular hydrogen protects against oxidative stress-induced SH-SY5Y neuroblastoma cell death through the process of mitohormesis
Murakami Y, Ito M, Ohsawa I. · PLOS ONE. 2017;12(5):e0176992.
What kind of evidence is this?
Preclinical
Controlled SH-SY5Y neuroblastoma-cell gas-exposure and oxidative-stress experiment
Other neurological conditions
Other forms
Not reported in this record.
Not reported in this record.
Methods at a glance
Human SH-SY5Y neuroblastoma cells cultured in glucose- or galactose-containing media.
Experiments were repeated two or three times with n=4–5 per group; selected protein measurements used n=5–6.
30-minute box fill followed by sealed exposures from one to 18 hours; oxidative-stress outcome timing varied by assay.
Pretreatment or post-treatment in sealed culture boxes with gas containing 1–50% H₂, principally 50% for up to 18 hours, around hydrogen-peroxide exposure.
50% H₂ with 10% O₂, 5% CO₂ and 35% N₂ — H₂ alone as the active gas component, not Brown's gas.
Boxes were filled at 1 L/min for 30 minutes and then sealed. At 50% H₂, the fill stream contained 500 mL/min H₂ and 100 mL/min O₂; these are filling flows, not continuous 18-hour exposure flows. Medium H₂ was 390±40 μM.
500 mL/min during the 30-minute box fill at 50% H₂; no continuous flow during sealed incubation.
100 mL/min during the box fill at 10% O₂; no continuous flow during sealed incubation.
Matched 10% O₂/5% CO₂/85% N₂ gas without H₂, multiple H₂ concentrations/durations and pretreatment versus post-treatment.
Outcomes and reported result
Cell death, mitochondrial membrane potential, ATP, oxygen consumption, mitochondrial DNA, glutathione/superoxide/ROS and Nrf2-pathway gene/protein expression.
H₂ pretreatment of at least three hours reduced later hydrogen-peroxide cell death and changed mitochondrial and Nrf2-linked measures. Post-treatment did not protect; mitochondrial DNA copy number did not change, DCFDA-indicated ROS did not increase, and the induced stress included lower glutathione and higher superoxide in one metabolic condition.
The complete free PMC article, figures and gas-system methods were checked for exact mixture and filling flow, concentration/duration comparisons, positive and null/adverse-direction findings, funding and conflicts.
A reported association, difference or mechanism is not automatically a clinical benefit. Null findings, outcome type, study design and precision all matter.
Cautions and applicability
Cancer-derived cell line, high gas concentrations, sealed-box exposure, small laboratory replicate counts, pretreatment paradigm and many mechanistic endpoints. Japanese public grants funded the work; authors declared no competing interests.
SH-SY5Y cells under induced oxidative stress; it does not establish neuroprotection or cancer treatment in animals or people.
Not reported in this record.
Not reported in this record.
Not reported in this record.
No single-study GRADE certainty rating is assigned. Certainty is assessed by important outcome across a complete eligible evidence set, not by attaching a final grade to one source.
Sources and record status
PMID: 28467497 · DOI: 10.1371/journal.pone.0176992
Free full article in PubMed Central.
Complete PMC article and PubMed metadata; full-text extraction checked 9 August 2026.
2026-08-10