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Y. Morizane, A. Thanos, G. Trichonas, X. Koufomichali, A. Manola, E. S. Gragoudas, J. W. Miller, B. Viollet, D. Vavvas; Regulation of Matrix Metalloproteinase 2 by AMP-Dependent Kinase. Invest. Ophthalmol. Vis. Sci. 2009;50(13):2927.
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Metalloproteinase (MMP) 2 is a gelatinase that degrades a broad range of collagens, elastin and fibronectin. MMP2 plays an important role in angiogenesis; however, the mechanisms regulating MMP2 activity are not fully understood.AMP-dependent Kinase (AMPK) is a primary sensor of cellular energy change. AMPK regulates energy homeostasis by switching off ATP-consuming biosynthetic pathways and protects cells from metabolic or nutritional stress. Recently, AMPK was reported to stimulate angiogenesis. We investigated whether AMPK regulates the activity of MMP2.
AMPK wild type (WT) and AMPK1-/-2-/- (KO) mouse embryonic fibroblasts (MEFs) were grown in Dulbecco’s modified Eagle’s medium / F12 with 10% heat inactivated fetal bovine serum. After culture for 24 hours in serum-free medium, MEFs were stimulated with interleukin-1β (IL1-β, 0.5-20 ng/ml) or tumor necrosis factor (TNF, 1-200 ng/ml). After 24 hours, media was evaluated by gelatin zymography normalized by protein concentration of cell extracts. Results of zymography were quantified using Image J software for relative area measurements. Data are expressed as a percent of the area obtained from the untreated control group.
In WT MEFs, both IL1-β (1 and 5 ng/ml) and TNF (5 and 10 ng/ml) significantly increased the activity of MMP2 in a dose-dependent manner (135%, p<0.05; 158%, p<0.01; 121%, p<0.05; 133%, p<0.01 respectively).Compared to the WT MEFs, MMP2 activity in KO MEFs was significantly lower when stimulated by IL-1β (5 ng/ml) (158% in WT vs 113% in KO, p<0.05) and further decreased after TNF (5 ng/ml) stimulation (121% in WT vs 88% in KO, p<0.05).
AMPK appears to increase activity of MMP2 following stimulation with inflammatory cytokines.
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