The effect of L-carnitine and acetyl-L-carnitine on the disappearance of DNA single-strand breaks in human peripheral blood lymphocytes.
DNA single-strand breaks (SSBs) and their disappearance during repair incubation were determined by alkaline filter elution in freshly isolated human peripheral blood lymphocytes (PBLs) after in vitro treatment with either the oxygen radical-generating system of xanthine oxidase (XOD) plus hypoxanthine (HYP) or the alkylating agent N-ethyl-N'-nitrosourea (ENU). The elution curves obtained with DNA from PBLs treated with XOD/HYP were markedly nonlinear, possibly as a result of a nonrandom induction of SSBs along the DNA strands. The disappearance of XOD/HYP-induced SSBs during the initial repair period was quite slow; only 20 +/- 7% (n = 6) of the induced SSBs had disappeared after a 2 1/2 h repair incubation. However, by 24 h the elution curves obtained with DNA from treated PBLs were indistinguishable from those obtained with DNA from nontreated control cells, indicating complete repair. Treatment of PBLs with ENU resulted in linear elution curves. Approximately 50% of the total amount of ENU-induced SSBs had disappeared within 1 h in PBLs from most donors; the additional SSBs were found to be persistent (Beorrigter, M.E.T.I., Mullaart, E., Berends, F., and Vijg, J. (1991) Induction and disappearance of DNA strand breaks and/or alkali-labile sites in human lymphocytes exposed to N-ethyl-N'-nitrosoureas. Carcinogenesis, 12, 77-82). Preincubation of PBLs with 5 mM L-carnitine, a trans-mitochondrial carrier of acetyl and long-chain acyl groups, or 5 mM acetyl-L-Carnitine, resulted in a more rapid disappearance of XOD/HYP-induced SSBs (48 +/- 23% and 48 +/- 30% respectively). Preincubation of PBLs with different doses of L-carnitine, before exposure to 0.5 mM ENU, increased SSB disappearance dependent on the dose and donor PBLs. In conclusion, these studies suggest that treatment with L-carnitine accelerates the disappearance of SSBs induced by oxygen radicals and alkylating agents.
Boerrigter
M E
ME
Division on Aging, Harvard Medical School, Boston, MA.
Franceschi
C
C
Arrigoni-Martelli
E
E
Wei
J Y
JY
Vijg
J
J
D000108:Acetylcarnitine / Q000494:pharmacology; D000328:Adult; D002331:Carnitine / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D004249:DNA Damage / Q000187:drug effects*; D004277:DNA, Single-Stranded / Q000187:drug effects*; D005038:Ethylnitrosourea / Q000633:toxicity; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000633:toxicity; D008214:Lymphocytes / Q000187:drug effects*; D013997:Time Factors; D014969:Xanthine Oxidase / Q000633:toxicity
D004277:DNA, Single-Stranded; D007042:Hypoxanthines; D019271:Hypoxanthine; D000108:Acetylcarnitine; D006861:Hydrogen Peroxide; D014969:Xanthine Oxidase; D005038:Ethylnitrosourea; D002331:Carnitine