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.
Study at a glance
Preclinical
Four-week-old male C57BL/6J mice exposed to chronic nicotine, with hydrogen-rich saline, vitamin C or vitamin E comparator treatments.
H₂-rich saline 6 mL/kg intraperitoneally each morning for three months alongside subcutaneous nicotine 4.5 mg/kg/day. · 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.
Daily treatment for three months.
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.
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.
What kind of evidence is this?
Preclinical
Randomized five-group controlled mouse experiment
Other molecular hydrogen research
Other forms
Information not yet classified
Some editorial classification fields are still pending. The source-reported outcomes and result are shown below; Hydrogenology does not infer a positive or negative signal from prose automatically.
Methods
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.
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.
Limitations 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.
No single-study GRADE certainty rating is assigned. Read how records and evidence assessments are prepared.
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.
10 August 2026
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