title: Ciprofloxacin has antifibrotic effects in scleroderma fibroblasts via             downregulation of Dnmt1 and upregulation of Fli1.
abstract: Systemic sclerosis (SSc) is characterized by fibrosis of the skin and internal organs. The present study was undertaken to examine the effects of ciprofloxacin, a fluoroquinolone antibiotic implicated in matrix remodeling, on dermal and lung fibroblasts obtained from SSc patients. Dermal and lung fibroblasts from SSc patients and healthy subjects were treated with ciprofloxacin. Western blotting was used to analyze protein levels and RT-PCR was used to measure mRNA expression. The pharmacologic inhibitor UO126 was used to block Erk1/2 signaling. SSc dermal fibroblasts demonstrated a significant decrease in collagen type I mRNA and protein levels after antibiotic treatment, while healthy dermal fibroblasts were less sensitive to ciprofloxacin, downregulating collagen only at the protein levels. Connective tissue growth factor (CCN2) gene expression was significantly reduced and matrix metalloproteinase 1 (MMP1) levels were enhanced after ciprofloxacin treatment to a similar extent in healthy and SSc fibroblasts. Ciprofloxacin induced Erk1/2 phosphorylation, and Erk1/2 blockade completely prevented MMP1 upregulation. However, Smad1 and Smad3 activation in response to TGFβ was not affected. The expression of friend leukemia integration factor 1 (Fli1), a transcriptional repressor of collagen, was increased after treatment with ciprofloxacin only in SSc fibroblasts, and this was accompanied by a decrease in the levels of DNA methyltransferase 1 (Dnmt1). Similar effects were observed in SSc-interstitial lung disease (ILD) lung fibroblasts. In summary, our study demonstrates that ciprofloxacin has antifibrotic actions in SSc dermal and lung fibroblasts via the downregulation of Dnmt1, the upregulation of Fli1 and induction of MMP1 gene expression via an Erk1/2-dependent mechanism. Thus, our data suggest that ciprofloxacin may be an attractive therapy for SSc skin and lung fibrosis.
authors.affiliation: Arthritis Center, Rheumatology, Boston University School of Medicine, Boston, MA 02118, USA. andabujor@yahoo.com
authors.forename: Andreea M
authors.initials: AM
authors.lastname: Bujor
authors.forename: Paul
authors.initials: P
authors.lastname: Haines
authors.forename: Cristina
authors.initials: C
authors.lastname: Padilla
authors.forename: Romy B
authors.initials: RB
authors.lastname: Christmann
authors.forename: Monica
authors.initials: M
authors.lastname: Junie
authors.forename: Percival D
authors.initials: PD
authors.lastname: Sampaio-Barros
authors.forename: Robert
authors.initials: R
authors.lastname: Lafyatis
authors.forename: Maria
authors.initials: M
authors.lastname: Trojanowska
mesh_terms: D064236:Cartilage Oligomeric Matrix Protein; D016022:Case-Control Studies; D002478:Cells, Cultured; D002939:Ciprofloxacin / Q000494:pharmacology*; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D000090602:Collagen Type I, alpha 1 Chain; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics* / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects*; D004353:Drug Evaluation, Preclinical; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D008168:Lung / Q000473:pathology; D017563:Lung Diseases, Interstitial / Q000188:drug therapy / Q000473:pathology; D020935:MAP Kinase Signaling System / Q000187:drug effects; D064235:Matrilin Proteins; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism; D051789:Proto-Oncogene Protein c-fli-1 / Q000235:genetics* / Q000378:metabolism; D012595:Scleroderma, Systemic / Q000188:drug therapy / Q000473:pathology*; D012867:Skin / Q000473:pathology; D015854:Up-Regulation / Q000187:drug effects*
chemical_list: D064236:Cartilage Oligomeric Matrix Protein; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D016326:Extracellular Matrix Proteins; C078433:FLI1 protein, human; D006023:Glycoproteins; D064235:Matrilin Proteins; D051789:Proto-Oncogene Protein c-fli-1; C577553:TSP5 protein, human; D002939:Ciprofloxacin; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; C000619380:DNMT1 protein, human; C522360:MMP1 protein, human; D020781:Matrix Metalloproteinase 1