J Korean Acad Periodontol.  2003 Jun;33(2):277-288. 10.5051/jkape.2003.33.2.277.

Nitric Oxide on the MMP-2 expression by human gingival fibroblasts

• Shin IS
• Yoon SO
• Chung HJ
• Koh JT

• KMID: 1999805
• DOI: http://doi.org/10.5051/jkape.2003.33.2.277

Abstract

It has been suggested that increased number and activity of phagocytes in periodontitis lesion results in a high degree of reactive oxygen species (ROS) such as superoxide anion, hydrogen peroxide, nitric oxide and peroxynitrite. There are few reports on the relationship between ROS and MMPs expressions in gingival fibroblast. We studied to elucidate whether and how ROS, especially nitric oxide affects the MMP expression. Human gingival fibroblasts and HT1080 cells (human fibrosarcoma sell line as reference) were grown in DMEM supplemented with 10 mM HEPES, 50 mg/L gentamicin, and 10% heat inactivated fetal bovine serum with addition of various reactive oxygen species (ROS). Culture media conditioned by cells were examined by gelatin zymography. HT1080 cells expressed proMMP-2 and proMMP-9, but human gingival fibroblasts (HGF) produced only proMMP-2. Hydrogen peroxide upregulated MMP-9 expression in HT1080 cells, whereas in human gingival fibroblast SNP treatment showed marked increase in MMP-2 level compared to other ROS. These results suggest that the effects of ROS on MMPs expressions are cell-type specific. RT-PCR for MMP-2 and TIMP-2 m-RNA were performed using total RNA from cultured cells under the influence various kinase inhibitors. In HT1080 cells, treatment with FPTI III (Ras processing inhibitor) and LY294002 (PI3-kinase inhibitor) resulted in inhibition of MMP-2 and MMP-9 expressions, suggesting that Ras/PI3-kinase pathway is important for MMPs expression in HT1080 cells. In gingival fibroblasts, treatment with FPTI III and PDTC (NFkB inhibitor) showed marked decrease in MMP-2 regardless of the of SNP, suggesting that Ras/NF-kB could be the key pathway for NO-induced MMP-2 expression in gingival fibroblasts. This study showed that ROS, especially nitric oxide, could be the critical mediator of periodontal disease progression through control of MMP-2 expression in gingival fibroblasts possibly via Ras/NF-kB pathway.

Keyword

NO; MMP-2; gingival fibroblast; Ras/NF-kB

MeSH Terms

Cells, Cultured
Culture Media
Fibroblasts*
Fibrosarcoma
Gelatin
Gentamicins
HEPES
Hot Temperature
Humans*
Hydrogen Peroxide
Matrix Metalloproteinases
Nitric Oxide*
Periodontal Diseases
Periodontitis
Peroxynitrous Acid
Phagocytes
Phosphotransferases
Reactive Oxygen Species
RNA
Superoxides
Tissue Inhibitor of Metalloproteinase-2
Culture Media
Gelatin
Gentamicins
HEPES
Hydrogen Peroxide
Matrix Metalloproteinases
Nitric Oxide
Peroxynitrous Acid
Phosphotransferases
RNA
Reactive Oxygen Species
Superoxides
Tissue Inhibitor of Metalloproteinase-2