Propolis decreases the expression of MMP-9 in fibroblasts and monocyte-derived macrophage in vitro / 解放军医学杂志

ABSTRACT

Objective To explore the potential mechanisms of therapeutic effect of propolis in the treatment of diabetic wound. Methods Human fibroblasts(Fb) and monocytic THP-1 cells were routinely cultured at 37℃ in an environment of 95% air and 5% CO2 in RPMI medium containing 11mmol/l glucose, 15mmol/l HEPES and 5% fetal calf serum. To a portion of culturing THP-1 monocytes phorbol myristate actetate (PMA; 2?10 -7mol/L) was added. After 24 hours when approximately 75% of the monocytes became macrophage which attached to the plate. Propolis was added to a culture system of human fibroblasts(Fb), THP-1 monocyte and THP-1 monocyte-derived macrophage(Mac) with different concentration of propolis(10, 50, 100, 200?g/ml) and different glucose(5mmol/L; 25mmol/L), For each experiment the cells were cultured for 48 hours in the presence of Propolis. Then the media were collected and the protein expression of matrix metalloproternase-9 (MMP-9) were determined by gelatin zymography using a MDPF- labeled gelatin as substrate. The media were loaded onto an 8% SDS PAGE gel containing MDPF labeled gelatin (1%) after electrophoresis, the gels were placed on a long wave ultra violet light box and photographed. The area degraded by the MMPs was observed as dark bands on a light background. The MMPs were quantified by measurement of the band area using the Phoretix_1D_Advanced Digital Science Electrophoresis Documentation and Analysis System. Results the high glucose has a tendency to increase the expression of MMP-9, The propolis can decreased the expression of MMP-9, and has a dependent on doses tendency. F test P