Detection of lysyl oxidase gene expression in rat skin during wound healing

Fushida-Takemura, H.; Fukuda, Michio; Maekawa, Naoki; Chanoki, Miyako; Kobayashi, Hiromi; Yashiro, Noriko; Ishii, Masamitsu; Hamada, Toshio; Otani, Shuzo; Ooshima, Akira

doi:10.1007/bf02505035

Cited by 37 publications

(24 citation statements)

References 22 publications

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“…33,45 Crosslinking of collagen fibers also occurs during wound healing responses and increased LOX expression has been reported to occur under such conditions. 46 The present observation that human skin fibroblasts carrying large amounts of the common deletion translate into structural and functional alterations into the extracellular matrix by virtue of LOX overexpression further supports the concept that skin aging processes and wound healing responses resemble each other. 47 Interestingly, Pyd crosslinks were mainly detected in the cytoplasm of the fibroblasts suggesting that LOX is active intracellularly during collagen gel contraction.…”

Section: Discussionsupporting

confidence: 74%

Functional Consequences of Mitochondrial DNA Deletions in Human Skin Fibroblasts

Majora

Wittkampf

Schuermann

et al. 2009

The American Journal of Pathology

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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 ␤-aminopropionitrile decreased the contraction difference between KSS and normal human fibroblast equivalents. Also, addition of the antioxidant N-tert-butyl-␣-phenylnitrone reduced the contraction difference by inhibiting collagen gel contraction in KSS fibroblasts, and both ␤-aminopropionitrile and N-tert-butyl-␣-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.

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Section: Discussionsupporting

confidence: 74%

Functional Consequences of Mitochondrial DNA Deletions in Human Skin Fibroblasts

Majora

Wittkampf

Schuermann

et al. 2009

The American Journal of Pathology

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“…Many studies have described up-or down-regulation of LOX in response to biological stimuli, but in many of these studies, the level of alteration between LOX mRNA and LOX activity was not consistent (51)(52)(53)(54)(55)(56)(57). In addition, the production of LOX mRNA in response to dermal injury has been demonstrated to be rapid (peaking after 3 days) (58). However, LOX enzyme activity has been shown to peak later (8 -10 days post-wound) (59), which suggests that the LOX pro-enzyme may be held in the ECM for a significant period of time before activation.…”

Section: Discussionmentioning

confidence: 99%

Cellular Fibronectin Binds to Lysyl Oxidase with High Affinity and Is Critical for Its Proteolytic Activation

Fogelgren

Polgár

Szauter

et al. 2005

Journal of Biological Chemistry

169

143

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Lysyl oxidase (LOX) is a copper-containing amine oxidase known to catalyze the covalent cross-linking of fibrillar collagens and elastin at peptidyl lysine residues. In addition, its involvement in cancer, wound healing, cell motility, chemotaxis, and differentiation reflect a remarkable functional diversity of LOX. To investigate novel mechanisms of LOX regulation and function, we performed a yeast two-hybrid screen to identify LOX-interacting proteins. Three overlapping positive clones were identified as C-terminal fragments of fibronectin (FN). Glutathione S-transferase pull-downs and solid phase binding assays confirmed this interaction. LOX binds to the cellular form of FN (cFN) with a dissociation constant (K d ) of 2.5 nM. This was comparable with our measured K d of LOX binding to tropoelastin (1.9 nM) and type I collagen (5.2 nM), but LOX demonstrated a much lower binding affinity for the plasma form of FN (pFN). Immunofluorescent microscopy revealed co-localization of FN and LOX in normal human tissues, where these proteins may interact in vivo. LOX enzymatic activity assays showed that cFN does not seem to be a substrate of LOX. However, cFN can act as a scaffold for enzymatically active 30-kDa LOX. Furthermore, in FN-null mouse embryonic fibroblasts, we observed dramatically decreased proteolytic processing of the 45-kDa LOX proenzyme to the 30-kDa active form, with a corresponding decrease in LOX enzyme activity. Our results suggest that the FN matrix may provide specific microenvironments to regulate LOX catalytic activity.Lysyl oxidase (LOX), 1 a copper-dependent amine oxidase, catalyzes the oxidative deamination of peptidyl lysine residues in collagen and elastin molecules to ␦-aminoadipic-␤-semialdehyde or allysine, which can then spontaneously condense with neighboring amino groups or other peptidyl aldehydes to form covalent cross-links in fibrillar collagens and elastin (1, 2). The covalent cross-linking of these extracellular matrix (ECM) proteins by LOX is essential to the formation of insoluble collagen and elastic fibers and for normal mammalian development (3, 4). In many human pathologies, including cardiovascular disease, fibrosis, and cancer, LOX expression and activity have been demonstrated to be altered as compared with normal conditions (1, 2).The human LOX protein is synthesized as a prepro-enzyme of 417 amino acids with an N-terminal signal peptide sequence for secretion. During protein trafficking in the Golgi, a copper atom is incorporated at the copper-binding site (residues 286 -296) (5-7), which allows the formation of a lysyl-tyrosylquinone cofactor derived from Tyr-355 and Lys-320 residues (8). After secretion into the extracellular space, proteolytic cleavage between residues Gly-168 and Asp-169 removes the N-terminal propeptide, yielding an active enzyme of 30 kDa (9 -11). This processing is mainly accomplished by the procollagen C-proteinase bone morphogenic protein-1 (BMP-1), and to a lesser degree, by mammalian Tolloid-like 1 protein (12, 13). However, it is not know...

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“…In recent years, various studies have clarified the mechanism by which connective tissue forms in normal and pathologic situations 7 . These studies have shown that formation of scar tissue in wounds produces an increase in the lysyl‐oxidase enzyme, which triggers cross‐linking of the aldehyde groups derived from the lysin amino acids, thus fomenting the maturation of collagen 7–9 . This enzyme is located outside the cells along the collagen and elastin fibers, and inside the cells in fibroblasts, endothelial cells, sweat and sebaceous glands, the erector pili muscle, and in some keratinocytes 10 .…”

Section: Discussionmentioning

confidence: 99%

Bleomycin in the Treatment of Keloids and Hypertrophic Scars by Multiple Needle Punctures

España

Solano

Quintanilla

2001

Dermatologic Surgery

101

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Background. The treatment of keloids and hypertrophic scars has been difficult and a recent French study showed that bleomycin has been useful in the treatment of these lesions. Objective. To determine the effectiveness and safety of bleomycin in the treatment of hypertrophic scars and keloids when this drug is administered through multiple superficial punctures. Methods. We applied bleomycin to keloids and hypertrophic scars in 13 patients using a multiple-puncture method on the surface of the skin. All patients were given bleomycin at a concentration of 1.5 IU/ml. Clinical response after treatment was classified according to the following scale: complete flattening (100%), highly significant flattening (>90%), or significant flattening (75-90%). Results. The clinical response was very positive in all cases: complete flattening in six cases, highly significant flattening in six cases, and significant flattening in one case. Two patients presented a recurrence as a small nodule 10 and 12 months after the last infiltration. Conclusions. These clinical findings show that administration of bleomycin in keloids and hypertrophic scars shows promise and needs further investigation.

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Detection of lysyl oxidase gene expression in rat skin during wound healing

Cited by 37 publications

References 22 publications

Functional Consequences of Mitochondrial DNA Deletions in Human Skin Fibroblasts

Functional Consequences of Mitochondrial DNA Deletions in Human Skin Fibroblasts

Cellular Fibronectin Binds to Lysyl Oxidase with High Affinity and Is Critical for Its Proteolytic Activation

Bleomycin in the Treatment of Keloids and Hypertrophic Scars by Multiple Needle Punctures

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