title: Functional consequences of mitochondrial DNA deletions in human skin fibroblasts: increased contractile strength in collagen lattices is due to oxidative stress-induced lysyl oxidase activity.
abstract: Deletions within the mitochondrial DNA (mtDNA) are thought to contribute to extrinsic skin aging. To study the translation of mtDNA deletions into functional and structural changes in the skin, we seeded human skin fibroblasts into collagen gels to generate dermal equivalents. These cells were either derived from Kearns-Sayre syndrome (KSS) patients, who constitutively carry large amounts of the UV-inducible mitochondrial common deletion, or normal human volunteers. We found that KSS fibroblasts, in comparison with normal human fibroblasts, contracted the gels faster and more strongly, an effect that was dependent on reactive oxygen species. Gene expression and Western blot analysis revealed significant upregulation of lysyl oxidase (LOX) in KSS fibroblasts. Treatment with the specific LOX inhibitor beta-aminopropionitrile decreased the contraction difference between KSS and normal human fibroblast equivalents. Also, addition of the antioxidant N-tert-butyl-alpha-phenylnitrone reduced the contraction difference by inhibiting collagen gel contraction in KSS fibroblasts, and both beta-aminopropionitrile and N-tert-butyl-alpha-phenylnitrone diminished LOX activity. These data suggest a causal relationship between mtDNA deletions, reactive oxygen species production, and increased LOX activity that leads to increased contraction of collagen gels. Accordingly, increased LOX expression was also observed in vivo in photoaged human and mouse skin. Therefore, mtDNA deletions in human fibroblasts may lead to functional and structural alterations of the skin.
authors: Institut fuer umweltmedizinische Forschung, Heinrich-Heine University Duesseldorf gGmbH, Auf'm Hennekamp 50, Duesseldorf, Germany.
authors: Marc
authors: M
authors: Majora
authors: Tanja
authors: T
authors: Wittkampf
authors: Bianca
authors: B
authors: Schuermann
authors: Maren
authors: M
authors: Schneider
authors: Susanne
authors: S
authors: Franke
authors: Susanne
authors: S
authors: Grether-Beck
authors: Ekkehard
authors: E
authors: Wilichowski
authors: Françoise
authors: F
authors: Bernerd
authors: Peter
authors: P
authors: Schroeder
authors: Jean
authors: J
authors: Krutmann
mesh_terms: D000629:Aminopropionitrile / Q000494:pharmacology; D000818:Animals; D002478:Cells, Cultured; D003094:Collagen; D003497:Cyclic N-Oxides / Q000494:pharmacology; D004249:DNA Damage; D004272:DNA, Mitochondrial / Q000235:genetics*; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000502:physiology*; D006801:Humans; D007625:Kearns-Sayre Syndrome / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008928:Mitochondria / Q000235:genetics; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D008249:Protein-Lysine 6-Oxidase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D017384:Sequence Deletion; D012867:Skin / Q000378:metabolism / Q000473:pathology; D015595:Skin Aging
chemical_list: D003497:Cyclic N-Oxides; D004272:DNA, Mitochondrial; D004791:Enzyme Inhibitors; D017382:Reactive Oxygen Species; D000629:Aminopropionitrile; C029217:phenyl-N-tert-butylnitrone; D003094:Collagen; D008249:Protein-Lysine 6-Oxidase