蜂胶对成纤维细胞、单核巨噬细胞系基质金属蛋白酶MMP-9表达的抑制作用

Propolis decreases the expression of MMP-9 in fibroblasts and monocyte-derived macrophage in vitro

目的探讨蜂胶对糖尿病伤口愈合的作用及其机制。方法采用荧光电泳法,检测不同葡萄糖浓度(5、25mmol/L)和不同蜂胶浓度(10、50、100、200μg/ml)处理的体外培养的人成纤维细胞及单核巨噬细胞系的基质金属蛋白酶MMP9蛋白的表达。结果高浓度葡萄糖(25mmol/L)状态下,MMP9表达增加,但与低浓度(5mmol/L)比较,差异无统计学意义。蜂胶可显著降低MMP9的表达,并呈剂量依赖性(P<0.05)。结论高糖能增加单核巨噬细胞系及成纤维细胞MMP9的表达,可能会影响糖尿病患者伤口的愈合。蜂胶可抑制这些细胞MMP9的表达,有益于细胞基质的积聚,从而有益于糖尿病患者伤口的愈合。

Objective To explore the potential mechanisms of therapeutic effect of propolis in the treatment of diabetic wound. Methads Human fibroblasts（Fb） and monocytic THP-1 cells were routinely cultured at 37℃ in an environment of 95% air and 5% CO2 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 differem concentration of propolis（10, 50, 100, 200μg/ml） and different glucose（Smmol/L; 25mmol/L）,For each experiment the ceils were cultured for 48 hours in the presence of Propoiis. 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. Themedia 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 hands on a light background. The MMPs were quantified by measurement of the hand 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〈0. 05. Condusion propolis significantly depress the expression of MMP-9, may play a role in wound healing by decreasing matrix turnover. That is appears to be more efficient in cells exposed to high glucose concentration suggest that propolis may improve wound healing in diabetes .