摘要
BACKGROUND: The high level of matrix metalloproteinase 9(MMP9) is thought to slow down the healing of diabetic foot ulcers. Whether it can influence the biological behaviors of skin fi broblasts and affect wound healing is still unclear. The present study aimed to observe changes in the biological behaviors of rat dermal fi broblasts induced by high expression of MMP9 and to clarify the possible mechanisms of wound healing for diabetic foot.METHODS: A cell model of skin f ibroblast with high expression of MMP9 was established by coculture of high glucose(22.0 mmol/L) and homocysteine(100 μmol/L). A control group was incubated with normal glucose(5.5 mmol/L). Realtime PCR, ELISA and gelatin zymography were used to detect the MMP9 mRNA, protein expression and activity of MMP9. Flow cytometry, CCK-8, ELISA assay, scratch test and transwell were used to detect cell proliferation, viability, collagen(hydroxyproline) secretion, horizontal migration and vertical migration of cells. The data were expressed as mean±SD. P value less than 0.05 was considered statistically signif icant.RESULTS: The expression of MMP9 mRNA, protein levels and the activity of MMP9 were much higher in the high MMP9 group than in the control group(7.05±1.02 vs. 1.00±0.00, 206.9±33.6 pg/mL vs. 40.4±5.9 pg/mL, and 1.47±0.13 vs. 0.57±0.12, respectively, P<0.01). The proportion of S-phase cells, proliferation index, cell viability, collagen(hydroxyproline) secretion, horizontal migration rate and the number of vertical migration cells were lower in the high MMP9 group than in the control group(P<0.01).CONCLUSION: Fibroblasts with a high expression of MMP9 decreased proliferation, activity, secretion and migration of collagens, suggesting that MMP9 may inhibit the biological behaviors of fi broblasts.
BACKGROUND： The high level of matrix metalloproteinase 9 （MMP9） is thought to slow down the healing of diabetic foot ulcers. Whether it can influence the biological behaviors of skin fibroblasts and affect wound healing is still unclear. The present study aimed to observe changes in the biological behaviors of rat dermal fibroblasts induced by high expression of MMP9 and to clarify the possible mechanisms of wound healing for diabetic foot. METHODS： A cell model of skin fibroblast with high expression of MMP9 was established by coculture of high glucose （22.0 mmol/L） and homocysteine （100 μmol/L）. A control group was incubated with normal glucose （5.5 mmol/L）. Realtime PCR, ELISA and gelatin zymography were used to detect the MMP9 mRNA, protein expression and activity of MMP9. Flow cytometry, CCK-8, ELISA assay, scratch test and transwell were used to detect cell proliferation, viability, collagen （hydroxyproline） secretion, horizontal migration and vertical migration of cells. The data were expressed as mean＋SD. P value less than 0.05 was considered statistically significant. RESULTS： The expression of MMP9 mRNA, protein levels and the activity of MMP9 were much higher in the high MMP9 group than in the control group （7.05±1.02 vs. 1.00±0.00,206.9±33.6 pg/mL vs. 40.4±5.9 pg/mL, and 1.47±0.13 vs. 0.57±0.12, respectively, P〈0.01）. The proportion of S-phase cells, proliferation index, cell viability, collagen （hydroxyproline） secretion, horizontal migration rate and the number of vertical migration cells were lower in the high MMP9 group than in the control group （P〈0.01）. CONCLUSION： Fibroblasts with a high expression of MMP9 decreased proliferation, activity, secretion and migration of collagens, suggesting that MMP9 may inhibit the biological behaviors of fibroblasts.
基金
supported by grants from the National Natural Science Foundation of China(81070660)
the Science and Technology Project Foundation of Guangdong Province(2008A030201012)
Medical Science and Technology Research Foundation of Guangdong Province(A2012183)
the Science and Technology Project Foundation of Guangdong Province(2009B091300128)
