Abstract：AIM: To observe the effect of different concentrations of glucose and homocysteine on the expression of MMP-9 in fibroblasts, and to investigate the relationship between homocysteine and diabetic foot.METHODS: Fibroblasts were obtained from the dermis of 1-day-old normal SD rat and cultured in DMEM containing 10% FBS. After deprivation of serum (using 0.5% FBS) for 24 h, the fibroblasts in 2-4 passages were cultured for 6 h under the conditions of normal glucose, high glucose and high glucose with high concentration of homocysteine, respectively. The expression of MMP-9 at mRNA and protein levels was assessed by RT-PCR and Western blotting, and the activity of MMP-9 was determined by gelatin zymography analysis. RESULTS: The expression of MMP-9 at mRNA and protein levels and activity of MMP-9 in high glucose (22 mmol/L) group were 1.15 folds, 1.59 folds and 1.34 folds of those in normal glucose group (P<0.05), respectively. The effect of high glucose became more pronounced when co-treated with high concentration of homocysteine (100 μmol/L), which were 1.25 folds, 2.63 folds and 2.52 folds of those in normal glucose group (P<0.05), respectively.CONCLUSION: The increase in the expression of MMP-9 is concentration-dependen with glucose content in the culture. High concentration of homocysteine promotes this process, suggesting that hypercysteinemia might play an important role in the expression of MMP-9 in fibroblast.
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