■&目标:氧化还原信号是伤口愈合过程中的关键调节剂。该信号通路可通过ECM有效地促进糖尿病性溃疡的发展或愈合。在这项研究中,研究了导致器官截肢的糖尿病溃疡患者组织中与氧化还原信号相关的细胞外基质结构。
■以糖尿病患者溃疡为病例组,非糖尿病肢体缺血为对照,采用病例对照设计。苏木精-伊红,三色,和弹性蛋白染色方法用于ECM的病理学评价。MDA,总硫醇,使用ELISA试剂盒测量SOD水平以评估氧化应激水平。此外,两组均采用ELISA试剂盒检测NO水平。使用具有SYBR-green测定的实时PCR检测基因MMP2、MMP9和HIF的表达水平。
■病理结果显示胶原蛋白和弹性蛋白纤维的厚度增加。在从糖尿病伤口组分离的组织中可见脂质萎缩。MAD量评价糖尿病溃疡患者脂质氧化程度明显高于对比组(p<0.01)。糖尿病溃疡组的硫醇水平明显低于对照组(p<0.0001)。分离自糖尿病溃疡的组织中金属蛋白酶2和9基因的表达低于对照组(p<0.0001)。而该组中HIF基因的表达高于对照组(p<0.0001)。
■在糖尿病伤口中,缺氧条件导致的HIF分泌有利于基质沉积并阻止蛋白酶活性,但如果缺氧持续,它可以导致ECM沉积,随后增加组织压力,由于更多的缺氧,胶原蛋白积累中胶原蛋白I与胶原蛋白III的比例增加,脂类萎缩并最终截肢。
BACKGROUND: & Objectives: Redox signaling is a critical regulator in the process of wound healing. This signaling pathway can be effective in the development or healing of diabetic ulcers through the ECM.In this study, the structure of extracellular matrix investigated in relation to redox signaling in the tissue of patients with diabetic ulcers that lead to organ amputation.
METHODS: The case-control design on diabetic patients ulcers as case group and non-diabetic limb ischemia as control were used.Hematoxylin-eosin, trichrome, and elastin staining methods were used for pathological evaluations of ECM. MDA, total thiol, and SOD levels were measured using ELISA kits to assess the oxidative stress level. Also, NO level was measured by using ELISA kits in both groups. Expression levels of genes MMP2, MMP9, and HIF were detected using real-time PCR with SYBR-green assay.
RESULTS: The pathological results showed an increase in the thickness of collagen and elastin fibers. Lipids atrophy was visible in the tissue isolated from the diabetic wound group. The amount of MAD to evaluate the level of lipid oxidation in patients with diabetic Ulcer was significantly higher than the control group(p < 0.01). Thiol level was significantly lower in the diabetic ulcer group than in the control group(p < 0.0001). The expression of metalloproteinases 2 and 9 genes in the tissues isolated from diabetic ulcers was lower than the control group(p < 0.0001). While the expression of the HIF gene in this group was higher than the control group(p < 0.0001).
CONCLUSIONS: In the diabetic wound, the HIF secretion due to hypoxic conditions is beneficial for matrix deposition and prevents protease activity, but if the hypoxia persists, it can lead to ECM deposition subsequently increases the tissue pressure, increases of the collagen I-to-collagen III ratio in collagen accumulation that due to more hypoxia , lipidsAtrophy and eventually amputation.