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、C 4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=0.000);A1、A2、B、C 4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。
Objective 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 bac- terial 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.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2017年第2期281-284,共4页
Chinese Journal of Experimental Surgery
基金
湖北省科技厅慢性创面及糖尿病足医学临床研究中心建设项目