Long-term treatment of hydrogen-rich saline abates testicular oxidative stress induced by nicotine in mice
Li S, Lu D, Zhang Y, Zhang Y. · Journal of Assisted Reproduction and Genetics. 2014;31(1):109-114.
What kind of evidence is this?
Preclinical
Randomized five-group controlled mouse experiment
Other molecular hydrogen research
Other forms
Not reported in this record.
Not reported in this record.
Methods at a glance
Four-week-old male C57BL/6J mice exposed to chronic nicotine, with hydrogen-rich saline, vitamin C or vitamin E comparator treatments.
60 mice randomized to five groups of n=12: control, nicotine, nicotine plus H₂-rich saline, nicotine plus vitamin C, or nicotine plus vitamin E.
Daily treatment for three months.
H₂-rich saline 6 mL/kg intraperitoneally each morning for three months alongside subcutaneous nicotine 4.5 mg/kg/day.
H₂ dissolved in physiological saline — H₂ only, not Brown's gas.
Hydrogen was dissolved for six hours at 0.4 MPa; saline was prepared weekly, stored without headspace at 4°C and its H₂ concentration was confirmed by gas chromatography. The article does not state a numerical dissolved-H₂ concentration.
Not applicable — intraperitoneal saline, not gas inhalation.
No O₂ was co-delivered as part of the intervention.
Nicotine plus ordinary saline, nicotine plus vitamin C, nicotine plus vitamin E, and non-nicotine control.
Outcomes and reported result
Testicular histology, epididymal sperm count and motility, serum and testicular testosterone, MDA, H₂O₂, nitrotyrosine, protein carbonyl and caspase-3 activity.
H₂-rich saline improved histological appearance, sperm count/motility and testosterone and reduced multiple oxidative and apoptotic measures versus nicotine plus saline. Vitamin C/E increases in sperm and testosterone were not significant; vitamin C did not reduce testicular H₂O₂, and neither vitamin significantly changed testicular nitrotyrosine, protein carbonyl or caspase-3.
The complete free PMC article was checked for randomization, all five groups, exact doses, H₂-saline preparation, positive and null comparator results, the authors' stated limitation, funding and available disclosures.
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
Young-mouse nicotine model, one sex, three-month treatment, surrogate reproductive and biochemical endpoints, no fertility or offspring endpoint, and no numerical H₂ concentration. The authors state that pituitary gonadotropins were not tested. No funding or conflict statement was identified in the article, so absence is not inferred.
Chronic nicotine exposure in young male mice; it does not establish a treatment for human infertility or tobacco-related reproductive injury.
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: 24221909 · DOI: 10.1007/s10815-013-0102-2
Free full article in PubMed Central.
Complete PMC article and PubMed metadata; full-text extraction checked 9 August 2026.
2026-08-10