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Zymographic analysis of gelatinases patterns in peripheral blood mononuclear cells| Abstract

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Zymographic analysis of gelatinases patterns in peripheral blood mononuclear cells

Abstract

Fatemeh Hajighasemi, Sakineh Hajighasemi

Gelatinases belongs to a large family of proteolytic enzymes named matrix metalloproteinases (MMPs) have a key role in inflammation and autoimmunity. In this study patterns of Gelatinase-A (MMP-2) and Gelatinase-B (MMP-9) activity in human peripheral blood mononuclear cells (hPBMCs) have been evaluated in vitro. HPBMCs were cultured in complete RPMI-1640 medium. Then the cells were seeded at a density of 10 5 cells/ml and were incubated with PMA (25ng/ml) or PHA (10 µg/ml) for 24 hours. Afterward the MMP-2 and MMP-9 activities in cell-conditioned media were evaluated by gelatin zymography. Statistical comparisons between groups were made by student T-test. PHA significantly increased MMP-9 activity in hPBMCs after 24 hour incubation time compared with untreated control cells. On the contrary, PMA extensively decreased MMP-9 activity in hPBMCs after 24 hour incubation time compared with untreated control cells. Besides hPBMCs did not show any MMP-2 activity and PHA/PMA had no effect on MMP-2 activity in hPBMCs compared with untreated control cells. According to the results of this study hPBMCs could exhibit MMP-9 activity. PHA and PMA had opposite effect on MMP-9 activity. PHA up-regulated while PMA down-regulated MMP-9 activity in hPBMCs. Thus, PHA/PMA stimulated hPBMCs, may provide valuable screening tools for MMP-9 enhancers/inhibitors and study of the regulatory mechanisms of MMP-9 activity.

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