Changes in biological behaviors of rat dermal fibroblasts induced by high expression of MMP9

Abstract

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 fi broblast 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 signifi cant. 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.

The effect of local hyperglycemia on skin cells in vitro and on wound healing in euglycemic rats

“…18 Existing literature was used when selecting the specific concentrations to represent these conditions in the experiments. 8,19,20 Deveci et al 21 showed that a high-glucose concentration (35 mM) induced apoptosis in keratinocytes in vitro. Our results showed that in vitro both skin cell types were able to tolerate hyperglycemic conditions well (up to 26 mM), and fibroblasts cultured in slightly hyperglycemic concentration (10 mM) exhibited increased proliferation in comparison to lower glucose concentrations.…”

Fibroblast-like synoviocyte metabolism in the pathogenesis of rheumatoid arthritis

“…Also, FLS secrete collagenases and metalloproteases (MMP), such as MMP-9, that start degrading cartilage ECM after activation [108]. Other MMP involved are MMP-13 [109], MMP-1 [110], MMP-2 [111], and MMP-3.…”

“…Several molecules and growth factors, such as PDGF, are essential for the formation of ECM-degrading invadosomal structures and cartilage invasion and activate several signaling pathways, including the PI3K/AKT pathway, known to be upstream of glucose metabolism [ 107 ]. Also, FLS secrete collagenases and metalloproteases (MMP), such as MMP-9, that start degrading cartilage ECM after activation [ 108 ]. Other MMP involved are MMP-13 [ 109 ], MMP-1 [ 110 ], MMP-2 [ 111 ], and MMP-3.…”

Transformation of fibroblast‐like synoviocytes in rheumatoid arthritis; from a friend to foe

“…PDGF is essential for the formation of hypertrophic FLS architecture and the extracellular matrix-degrading invadosome, invasion to cartilage, via activation of several signaling pathways, particularly the PI3K/AKT pathway [ 86 ]. Also, the intimal lining has been identified as the primary source of MMPs in RA, and is able to active FLSs with the secretion of collagenases and metalloproteinases, such as MMP-2 and MMP-9, and initiate the destruction of ECM and cartilage [ 87 , 88 ].…”