Journal of Periodontal and Implant Science

Korean Academy of Periodontology (대한치주과학회)
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Nitric Oxide on the MMP-2 expression by human gingival fibroblasts

치은섬유아세포의 MMP 발현에 대한 Nitric Oxide의 영향

  • Shin, In-Sik (Department of Periodontology, Chonnam National University) ;
  • Yoon, Sang-Oh (Department of Biological Sciences, Korea Advance Institute of Science and Technology) ;
  • Chung, Hyun-Ju (Department of Periodontology, Chonnam National University) ;
  • Koh, Jung-Tae (Department of Dental Pharmacology, College of Dentistry & Dental Science Research Institute, Chonnam National University)
  • 신인식 (전남대학교 치과대학 치주과학교실) ;
  • 윤상오 (한국과학기술원 생물과학실) ;
  • 정현주 (전남대학교 치과대학 치주과학교실) ;
  • 고정태 (전남대학교 치과대학 치과약리학교실, 치의학연구소)
  • Published : 2003.06.30
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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 HTl080 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/P13-kinase pathway is important for MMPs expression in HT1080 cells. In gingival fibroblasts, treatment with FPTI III and PDTC (NF-kB 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.

Keywords

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