title: High levels of γ-H2AX foci and cell membrane oxidation in adolescents with type 1 diabetes.
abstract: Oxidative stress caused by an excess of free radicals is implicated in the pathogenesis and development of type 1 diabetes mellitus (T1DM) and, in turn, it can lead to genome damage, especially in the form of DNA double-strand break (DSB). The DNA DSB is a potentially carcinogenic lesion for human cells. Thus, we aimed to evaluate whether the level of oxidative stress was increased in peripheral blood lymphocytes of a group of affected adolescents. In 35 T1DM adolescents and 19 healthy controls we assessed: (1) spontaneous and H2O2-induced oxidation of cell membrane using a fluorescence lipid probe; (2) spontaneous and LPS-induced expression of iNOS protein and indirect NO determination via cytofluorimetric analysis of O2(-); (3) immunofluorescent detection of the basal level of histone H2AX phosphorylation (γ-H2AX foci), a well-validated marker of DNA DSB. In T1DM, the frequencies of oxidized cells, both spontaneous and H2O2-induced (47.13±0.02) were significantly higher than in controls (35.90±0.03). Patients showed, in general, both a reduced iNOS expression and production of NO. Furthermore, the level of spontaneous nuclear damage, quantified as γ-H2AX foci, was markedly increased in T1DM adolescents (6.15±1.08% of γ-H2AX(+) cells; 8.72±2.14 γ-H2AXF/n; 9.26±2.37 γ-H2AXF/np), especially in females. In the present study, we confirmed the role that oxidative stress plays in the disease damaging lipids of cell membrane and, most importantly, causing genomic damage in circulating white blood cells of affected adolescents. This also indicates that oxidative stress can affect several tissues in the body. However, although the observed DNA damage is a clear indication that the proper DNA repair mechanisms are activated, the risk for young T1DM subjects of developing not only cardiovascular complications but also some type of cancer cannot be ruled out. In this view, females, probably due to hormonal imbalance typical of adolescence, might represent a more susceptible population.
authors.affiliation: Unità di Genetica, Dipartimento di Biologia, Pisa University, Pisa, Italy.
authors.forename: Caterina
authors.initials: C
authors.lastname: Giovannini
authors.affiliation: Sezione di Patologia Sperimentale, Dipartimento di Ricerca Traslazionale e delle Nuove Tecnologie in Medicina e Chirurgia, Pisa University, Pisa, Italy.
authors.forename: Simona
authors.initials: S
authors.lastname: Piaggi
authors.affiliation: Unità di Endocrinologia Pediatrica e Diabete, Dipartimento di Medicina Clinica e Sperimentale Pisa University, Pisa, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Federico
authors.affiliation: Unità di Genetica, Dipartimento di Biologia, Pisa University, Pisa, Italy. Electronic address: roberto.scarpato@unipi.it.
authors.forename: Roberto
authors.initials: R
authors.lastname: Scarpato
mesh_terms: D000293:Adolescent; D016022:Case-Control Studies; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D004249:DNA Damage; D003922:Diabetes Mellitus, Type 1 / Q000235:genetics* / Q000378:metabolism*; D005260:Female; D006657:Histones / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D015162:Micronucleus Tests; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000502:physiology
chemical_list: C468783:H2AX protein, human; D006657:Histones
keywords: Diabetes; Peripheral lymphocytes; iNOS; γ-H2AX