摘要目的 观察在不同细菌负荷的糖尿病创面中基质金属蛋白酶-9(MMP-9)及基质金属蛋白酶抑制剂-1(TIMP-1)表达的变化.方法 实验分为4组,19例糖尿病足创面患者为A组,其中7例有细菌生长的糖尿病创面患者为A1组,12例无细菌生长的糖尿病创面患者为A2组,6例糖尿病患者为B组,7例正常人为C组,手术中分别获取活组织标本,A组取活检进行细菌培养与计数,用实时荧光定量聚合酶链反应(FQ-PCR)分别检测组织中MMP-9及TIMP-1的基因表达,应用Western blot法验证组织中MMP-9及TIMP-1在蛋白水平的表达.结果 A1、A2、B、C4组中MMP-9 mRNA相对表达量分别为1.136 ±0.116、0.601±0.080、0.391 ±0.088、0.134±0.057,4组之间的差异呈下降趋势(P =O.000);A1、A2、B、C4组中TIMP-1 mRNA的相对表达量分别为1.013±0.086、2.570±0.845、4.878 ±0.829、7.294±1.551,4组之间的差异呈上升趋势(P=0.000).Western blot检测结果与FQ-PCR结果一致.结论 血糖升高改变皮肤组织中MMP-9及TIMP-1的表达,细菌负荷大的糖尿病创面MMP-9表达增加,而TIMP-1表达降低,可能使MMP-9/TIMP-1平衡失调,从而影响糖尿病创面的愈合.

abstractsObjective To observe the expression levels of matrix metalloproteinase (MMP)-9 and tissue inhibitor of metalloproteinase (TIMP)-1 in different bacterial colonization diabetic wound.Methods Included patients were divided into four groups:group A,19 patients with diabetic foot wound,including group A1,7 patients with bacterial growth on the wound,and group A2,12 patients without bacterial growth on the wound;group B,6 diabetic patients;group C,7 normal controls.Skin tissues in all four groups were taken surigically.Bacterial culture and counting were done in group A.The MMP-9 and TIMP-1 gene expression in skin tissue was detected by real-time fluorescent quantitative polymerase chain reaction (FQ-PCR),and Western blotting was used to detect the expression of MMP-9 and TIMP-1 proteins.Results The relative expression of MMP-9 in groups A1,A2,B and C was 1.136 ± 0.116,0.601 ±0.080,0.391 ±0.088 and 0.134 ±0.057 respectively (P =0.000 for all).The relative expression levels of TIMP-1 in groups A1,A2,B and C were 1.013 ±0.086,2.570 ±0.845,4.878 ± 0.829 and 7.294 ± 1.551 respectively (P =0.000 for all).The results from Western blotting were consistent with those from RT-PCR.Conclusion Hyperglycemia altered the expression of MMP-9 and TIMP-1 in the skin tissue.Heavy bacterial colonization in diabetic wound increased the expression of MMP-9,and decreased the TIMP-1 expression,which may disturb the balance of MMP-9/TIMP-1,affecting wound healing.