title: Anesthetic Isoflurane Induces DNA Damage Through Oxidative Stress and p53 Pathway.
abstract: DNA damage is associated with aging and neurological disorders, including Alzheimer's disease. Isoflurane is a commonly used anesthetic. It remains largely unknown whether isoflurane induces DNA damage. Phosphorylation of the histone protein H2A variant X at Ser139 (γH2A.X) is a marker of DNA damage. We therefore set out to assess the effects of isoflurane on γH2A.X level in H4 human neuroglioma cells and in brain tissues of mice. Oxidative stress, caspase-activated DNase (CAD), and the p53 signaling pathway are involved in DNA damage. Thus, we determined the interaction of isoflurane with reactive oxygen species (ROS), CAD, and p53 to illustrate the underlying mechanisms. The cells were treated with 2 % isoflurane for 3 or 6 h. The mice were anesthetized with 1.4 % isoflurane for 2 h. Western blot, immunostaining and live cell fluorescence staining were used in the experiments. We showed that isoflurane increased levels of γH2A.X, cleaved caspase-3, and nucleus translocation of CAD and decreased levels of inhibitor of CAD (ICAD) and p53. Isoflurane enhanced the nucleus level of γH2A.X. Moreover, caspase inhibitor Z-VAD and ROS generation inhibitor N-acetyl-L-cysteine (NAC) attenuated the isoflurane-induced increase in γH2A.X level. However, NAC did not significantly alter the isoflurane-induced reduction in p53 level. Finally, p53 activator (actinomycin D) and inhibitor (pifithrin-α) attenuated and potentiated the isoflurane-induced increase in γH2A.X level, respectively. These findings suggest that isoflurane might induce DNA damage, as represented by increased γH2A.X level, via induction of oxidative stress and inhibition of the repair of DNA damage through the p53 signaling pathway.
authors.affiliation: Department of Anesthesiology, Peking University Third Hospital, 49 North Garden Rd., Haidian District, Beijing, 100191, China.|Geriatric Anesthesia Research Unit, Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital and Harvard Medical School, 149 13th St., Room 4310, Charlestown, MA, 02129, USA.
authors.forename: Cheng
authors.initials: C
authors.lastname: Ni
authors.affiliation: Geriatric Anesthesia Research Unit, Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital and Harvard Medical School, 149 13th St., Room 4310, Charlestown, MA, 02129, USA.|Department of Anesthesiology, Shanghai Tenth People's Hospital, Tongji University, Shanghai, 200072, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Li
authors.affiliation: Geriatric Anesthesia Research Unit, Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital and Harvard Medical School, 149 13th St., Room 4310, Charlestown, MA, 02129, USA.
authors.forename: Yuanlin
authors.initials: Y
authors.lastname: Dong
authors.affiliation: Department of Anesthesiology, Peking University Third Hospital, 49 North Garden Rd., Haidian District, Beijing, 100191, China.
authors.forename: Xiangyang
authors.initials: X
authors.lastname: Guo
authors.affiliation: Geriatric Anesthesia Research Unit, Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital and Harvard Medical School, 149 13th St., Room 4310, Charlestown, MA, 02129, USA.
authors.forename: Yiying
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Geriatric Anesthesia Research Unit, Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital and Harvard Medical School, 149 13th St., Room 4310, Charlestown, MA, 02129, USA. zxie@mgh.harvard.edu.
authors.forename: Zhongcong
authors.initials: Z
authors.lastname: Xie
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D018685:Anesthetics, Inhalation / Q000494:pharmacology*; D000818:Animals; D052160:Benzothiazoles / Q000494:pharmacology; D001921:Brain / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D003600:Cytosol / Q000378:metabolism; D004249:DNA Damage; D003609:Dactinomycin / Q000494:pharmacology; D003851:Deoxyribonucleases / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D006657:Histones / Q000378:metabolism; D006801:Humans; D007530:Isoflurane / Q000494:pharmacology*; D051379:Mice; D008954:Models, Biological; D009842:Oligopeptides / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects*; D014050:Toluene / Q000031:analogs & derivatives / Q000494:pharmacology; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
chemical_list: D018685:Anesthetics, Inhalation; D052160:Benzothiazoles; C468783:H2AX protein, human; D006657:Histones; D009842:Oligopeptides; D016159:Tumor Suppressor Protein p53; C517629:benzyloxycarbonyl-valyl-alanyl-aspartic acid; D003609:Dactinomycin; D014050:Toluene; D007530:Isoflurane; C121565:pifithrin; D003851:Deoxyribonucleases; C109663:caspase-activated deoxyribonuclease; D053148:Caspase 3; D000111:Acetylcysteine
keywords: Anesthesia; Caspase-3; DNA damage; ROS; p53