Hydrogen-rich saline attenuates isoflurane-induced caspase-3 activation and cognitive impairment via inhibition of isoflurane-induced oxidative stress, mitochondrial dysfunction, and reduction in ATP levels
Li C, Hou L, Chen D, Lin F, Chang T, Li M, Zhang L, Niu X, Wang H, Fu S, Zheng J. · American Journal of Translational Research. 2017;9(3):1162-1172.
Study at a glance
Preclinical
H4 human neuroglioma cells overexpressing amyloid precursor protein and eight-month-old wild-type C57BL/6J mice.
Cells received 300 µmol/L H₂-rich saline 30 minutes before 2% isoflurane for three or six hours. Mice received 5 mL/kg H₂-rich saline intraperitoneally 30 minutes before 1.4% isoflurane in 100% O₂. · H₂ was dissolved in 0.9% saline for six hours at 0.4 MPa; weekly preparations were kept at or above 0.6 mmol/L. Cell treatment used 300 µmol/L; mouse dose was 5 mL/kg.
Cell exposures lasted three or six hours; mouse anesthesia was reported as two hours in methods and the figure caption, with fear-conditioning tests at 30/90 minutes, 48 hours and seven days.
H₂-rich saline attenuated isoflurane-associated caspase-3 activation, ROS and mitochondrial/ATP changes and improved two selected fear-conditioning findings: contextual freezing at day 7 and tone freezing at day 2. Saline alone did not significantly change full-length caspase-3, membrane potential or ATP; several assay descriptions report no baseline difference.
Cell-line and small mouse experiments, no H₂ dose-response, no in-vivo caspase-3 measurement, only one behavioral paradigm and an internal inconsistency: the methods/figure report two hours of anesthesia while one results sentence says three hours. Public and foundation grants were reported; no explicit conflict statement was identified, so absence is not inferred.
What kind of evidence is this?
Preclinical
Cell-culture mechanistic experiments plus randomized four-condition mouse fear-conditioning experiment
Other neurological conditions
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
H4 human neuroglioma cells overexpressing amyloid precursor protein and eight-month-old wild-type C57BL/6J mice.
Cell assays generally used six independent experiments per condition. Mice were randomized by weight and sex to four conditions with n=10 per group (40 mice total).
Cell exposures lasted three or six hours; mouse anesthesia was reported as two hours in methods and the figure caption, with fear-conditioning tests at 30/90 minutes, 48 hours and seven days.
Cells received 300 µmol/L H₂-rich saline 30 minutes before 2% isoflurane for three or six hours. Mice received 5 mL/kg H₂-rich saline intraperitoneally 30 minutes before 1.4% isoflurane in 100% O₂.
H₂ dissolved in saline — H₂ only, not Brown's gas. Isoflurane was delivered with oxygen, but H₂ was not inhaled.
H₂ was dissolved in 0.9% saline for six hours at 0.4 MPa; weekly preparations were kept at or above 0.6 mmol/L. Cell treatment used 300 µmol/L; mouse dose was 5 mL/kg.
Saline with or without isoflurane in cells and mice, producing four conditions in each experiment.
Outcomes and reported result
Caspase-3 activation, ROS, mitochondrial permeability and membrane potential, ATP, and contextual/tone fear-conditioning freezing time.
H₂-rich saline attenuated isoflurane-associated caspase-3 activation, ROS and mitochondrial/ATP changes and improved two selected fear-conditioning findings: contextual freezing at day 7 and tone freezing at day 2. Saline alone did not significantly change full-length caspase-3, membrane potential or ATP; several assay descriptions report no baseline difference.
The complete free PMC article was checked for cell and animal allocation, H₂ preparation/concentrations, mouse dose, oxygen flow, positive and null outcomes, internal duration inconsistency, limitations, funding and available disclosures.
A reported association, difference or mechanism is not automatically a clinical benefit.
Limitations and applicability
Cell-line and small mouse experiments, no H₂ dose-response, no in-vivo caspase-3 measurement, only one behavioral paradigm and an internal inconsistency: the methods/figure report two hours of anesthesia while one results sentence says three hours. Public and foundation grants were reported; no explicit conflict statement was identified, so absence is not inferred.
Isoflurane-exposed cells and mice; it does not establish prevention of postoperative cognitive problems in humans.
No single-study GRADE certainty rating is assigned. Read how records and evidence assessments are prepared.
Sources and record status
PMID: 28386342
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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