engleski

EFFECTS OF CIGARETTE SMOKE EXTRACT ON INDUCTION OF TIMP-1 AND MMP-9 IN HUMAN A549 CELL LINE

Background: Chronic obstructive pulmonary disease (COPD) is the most frequent chronic respiratory disease with the progressive course. It is characterised with the development of local and presumably systemic inflammation. Cigarette smoking is the main etiologic factor causing the inflammatory response in COPD patients. MMP-9 can play an important role in inflammation and lung tissue remodelling owing to its gelatinolitic activity. TIMP-1 is known as an inhibitor of matrix metalloproteinases, including MMP-9. In this study the effect of cigarette smoke extract (CSE) on induction of TIMP-1 and MMP-9 in human A549 cell line was examined. Materials and Methods: Human A549 cell line (lung carcinoma epithelial cells) were cultured in F-12 medium supplemented with 10 % FBS, 1 % GlutaMAX and 1 % Antibiotic-Antimycotic. The cigarette smoke extract was freshly prepared at the day of use by slowly bubbling the cigarette smoke through the F-12 medium. Cells (2 x 106 / well) were treated with different concentrations of CSE for 4h and 24h at 37°C. Supernatant was collected and frozen at -20°C until determination. TIMP-1 and MMP-9 were determined with the ELISA kit. Results: CSE induces the release of TIMP-1 from A549 cells after 24h incubation. Basal production of MMP-9 in A549 cells is very low and there was no induction of MMP-9 with any concentration of CSE. Conclusions: Human A549 cells stimulated with CSE release high concentrations of TIMP-1. In contrast to our expectations, CSE activated A549 cells do not release MMP-9. Further examination is needed in order to get more significant results.

A549, cigarette smoke, COPD, MMP-9, TIMP-1

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