Effect of transforming growth factor-beta1, interleukin-6, and interferon-gamma on the expression of type I collagen, heat shock protein 47, matrix metalloproteinase (MMP)-1 and MMP-2 by fibroblasts from normal gingiva and hereditary gingival fibromatosis.
BACKGROUND
Increased collagen and extracellular matrix deposition within the gingiva is the main characteristic feature of hereditary gingival fibromatosis (HGF). To date, it is not well established if these events are a consequence of alterations in the collagen and other extracellular matrix molecules synthesis or disturbances in the homeostatic equilibrium between synthesis and degradation of extracellular matrix molecules. Cytokines are important regulators of expression of the profibrogenic genes, including type I collagen and its molecular chaperone heat shock protein (Hsp)47 and proteolytic enzymes degrading extracellular matrix such as matrix metalloproteinases-1 and -2 (MMP-1 and MMP-2).


METHODS
In this study, we analyzed the expression and production of type I collagen, Hsp47, MMP-1, and MMP-2 in normal gingiva (NG) and HGF fibroblasts, and investigated the effects of transforming growth factor-beta1 (TGF-beta1), interleukin-6 (IL-6) and interferon-gamma (IFN-gamma) on the expression of these genes by NG and HGF fibroblasts.


RESULTS
Our results obtained from semi-quantitative reverse transcription-polymerase chain reactions (RT-PCR), Western blots, enzyme-linked immunosorbent assays (ELISA), and enzymographies clearly demonstrated that the expression and production of type I collagen and Hsp47 were significantly higher in fibroblasts from HGF than from NG, whereas MMP-1 and MMP-2 expression and production were lower in fibroblasts from HGF patients. Addition of TGF-beta1 and IL-6, which are produced in greater amounts by HGF fibroblasts, promoted an increase in type I collagen and Hsp47 and a decrease in MMP-1 and MMP-2 expression. IFN-gamma reduced both type I collagen and Hsp47 expression, whereas it had a slight effect on the expression of MMP-1 and MMP-2.


CONCLUSIONS
These patterns of expression and production suggest that enhanced TGF-beta1 and IL-6 production simultaneously increase the synthesis and reduce the proteolytic activities of fibroblasts from patients with HGF, which may favor the accumulation of extracellular matrix observed in patients with this condition.
Martelli-Junior
H
H
Discipline of Oral Pathology, University of Campinas Dental School, Piracicaba, São Paulo, Brazil.
Cotrim
P
P
Graner
E
E
Sauk
J J
JJ
Coletta
R D
RD
D000328:Adult; D000704:Analysis of Variance; D018929:Cell Culture Techniques; D024042:Collagen Type I / Q000187:drug effects*; D005260:Female; D005347:Fibroblasts / Q000187:drug effects*; D005351:Fibromatosis, Gingival / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D005881:Gingiva / Q000187:drug effects*; D050958:HSP47 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000187:drug effects*; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D015850:Interleukin-6 / Q000494:pharmacology*; D008297:Male; D020781:Matrix Metalloproteinase 1 / Q000187:drug effects*; D020778:Matrix Metalloproteinase 2 / Q000187:drug effects*; D018709:Statistics, Nonparametric; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
D024042:Collagen Type I; D050958:HSP47 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D015850:Interleukin-6; C494660:SERPINH1 protein, human; D016212:Transforming Growth Factor beta; D007371:Interferon-gamma; D020778:Matrix Metalloproteinase 2; D020781:Matrix Metalloproteinase 1