目的:建立高分泌基质金属蛋白酶9(matrix metalloproteinase 9,MMP9)的皮肤成纤维细胞模型,为探讨MMP9在糖尿病足伤口愈合中的作用提供细胞模型。方法:用不同浓度的糖和同型半胱氨酸培养皮肤成纤维细胞,检测各组细胞MMP9 mRNA及蛋白表达水平和活性,筛选出MMP9表达量最高的组。结果:(1)高糖+高同型半胱氨酸组MMP9mRNA表达量(7.05±1.02)显著高于正糖组和高糖组(P均〈0.01);(2)高糖+高同型半胱氨酸组MMP9蛋白表达量(206.9±33.6)pg/ml明显高于正糖组和高糖组(P均〈0.01);(3)高糖+高同型半胱氨酸组MMP9蛋白活性(1.47±0.13)高于正糖组和高糖组(P均〈0.01)。结论:高糖+高同型半胱氨酸联合培养成功建立体外高分泌MMP9皮肤成纤维细胞模型。
Objective:To establish a cell model of skin fibroblast with high expression of matrix metalloproteinase 9(MMP9) for studying its roles in wound healing of diabetes foots.Methods: Different concentrations of sugar and homocysteine were uesd to cultured skin fibroblasts.The expression levels of MMP9 mRNA and protein,and the activity of MMP9 protein were detected in each group.Results:(1) The expression level of MMP9 mRNA in high glucose + high homocysteine group(7.05±1.02) was significantly higher than those in the normal glucose group and high glucose group(respectively,P0.01).(2) The expression level of MMP9 protein in high glucose + high homocysteine group(206.9±33.6) pg/ml was significantly higher than those in the normal glucose group and high glucose group(respectively,P0.01).(3) The activity of MMP9 protein in high glucose + high homocysteine group(1.47±0.13) was significantly higher than those in the normal glucose group and high glucose group(respectively,P0.01).Conclusion: The cell model of skin fibroblast with high expression of MMP9 was successfully established in vitro by high glucose + high homocysteine co-culture.