Mechanistic study of Small-molecular Therapy in diabetic Wound Healing

导致截肢的糖尿病足溃疡是一个主要的健康问题，影响到3000万糖尿病患者中的20%。 美国的病人。目前的治疗方案成功有限，截肢率仍然很高。因此，我们认为， 理解具有翻译潜力的化合物的分子机制是使 伤口护理协议的突破内皮细胞（EC）是不可或缺的细胞成分， 伤口血管生成然而，糖尿病患者的EC功能受损。两个人的共同表述 膳食化合物-反式白藜芦醇（tRES）和橙皮素（HESP）-改善葡萄糖代谢谱， 通过诱导谷胱甘肽酶1（GLO1）的基因表达来改善超重和肥胖受试者的动脉功能， 一种在糖酵解过程中解毒反应性代谢物并保护细胞免受糖基化压力的酶。我们 初步数据表明，tRES + HESP改善了糖尿病动物的伤口愈合， 表情然而，它的作用可能远远超过诱导GLO1表达，因为tRES + HESP处理的细胞中， 内皮细胞产生许多促血管生成因子，包括血管生成素-1（ANGPT1），其在血管生成中起重要作用。 血管生成因此，确定在血管生成中由tRES + HESP调节的蛋白质是至关重要的 和组织修复。本项目的目标是填补tRES + HESP在救援中作用的知识空白 我们的长期目标是开发糖尿病的治疗策略， 伤口修复我们假设tRES + HESP增强血管生成并改善糖尿病伤口愈合 通过增强GLO1和一种有效的促血管生成因子ANGPT1的表达， 糖尿病伤口修复关键途径中其他蛋白质的变化。目的1：分子鉴定 tRES + HESP诱导的体外人真皮微血管内皮细胞中蛋白质的变化。次级目标 1：确定tRES + HESP在体外可以拯救糖尿病内皮细胞功能的程度。次级目标2： 确定ANGPT1在tRES + HESP诱导的体外血管生成中的重要性。子目标3：发现新蛋白质 和负责tRES + HESP治疗在体外内皮细胞功能中的益处的途径， 最先进的蛋白质组学目的2：确定tRES + HESP的治疗潜力及其基础分子 在体内慢性糖尿病伤口中的机制。子目标1：确定tRES + HESP对伤口的疗效 在db/db小鼠中新开发的糖尿病慢性伤口模型中的愈合。次级目标2：确定 ANGPT1在tRES + HESP诱导的体内伤口愈合改善中的作用。子目标3：发现新蛋白质 和负责tRES + HESP治疗在体内糖尿病伤口修复中的益处的途径， 最先进的蛋白质组学拟议研究的结果将决定局部应用的疗效 该公式，tRES + HESP，在糖尿病伤口愈合，并将揭示其潜在的分子机制， 有益效果。由于tRES + HESP尚未被FDA批准用于治疗糖尿病伤口愈合， 这些结果可促进FDA批准该共制剂用于糖尿病伤口治疗。

Diabetic foot ulcers that lead to amputations are a major health problem affecting ~20% of the 30 million diabetic patients in the US. The current regimen has limited success, and the amputation rates remain high. Therefore, understanding molecular mechanisms for compounds with translational potential is a crucial step toward making a breakthrough in wound care protocols. Endothelial cells (ECs) are indispensable cellular components for wound angiogenesis. However, EC functions are impaired in patients with diabetes. The coformulation of two dietary compounds - Trans-resveratrol (tRES) and hesperetin (HESP) - improves glucose metabolic profile and arterial function in overweight and obese subjects through inducing the gene expression of glyoxalase 1 (GLO1), an enzyme that detoxifies reactive metabolites during glycolysis and protects cells against glycation stress. Our pilot data indicated that tRES+HESP improved wound healing in diabetic animals with an increase in GLO1 expression. However, its effects are likely far beyond inducing GLO1 expression because tRES+HESP treated ECs produced many pro-angiogenic factors, including angiopoietin-1 (ANGPT1) that plays an essential role in angiogenesis. Therefore, it is critical to determine proteins that are regulated by tRES+HESP in angiogenesis and tissue repair. The objective of this project is to fill the knowledge gap of the role of tRES+HESP in rescuing the disrupted angiogenesis in diabetes, and our long-term goal is to develop therapeutic strategies for diabetic wound repair. We hypothesize that tRES+HESP augments angiogenesis and improves diabetic wound healing through enhancing the expression of GLO1 and a potent pro-angiogenic factor, ANGPT1, and through novel changes in additional proteins in pathways critical to diabetic wound repair. Aim 1: Identification of molecular pathways and protein changes induced by tRES+HESP in human dermal microvascular ECs in vitro. Sub-aim 1: Determine to what extent tRES+HESP can rescue diabetic endothelial cell function in vitro. Sub-aim 2: Determine how vital ANGPT1 is in tRES+HESP-induced angiogenesis in vitro. Sub-aim 3: Discover new proteins and pathways responsible for the benefit of tRES+HESP treatment in endothelial cell function in vitro using state- of-the-art proteomics. Aim 2: Determine the therapeutic potential of tRES+HESP and its underlying molecular mechanisms in chronic diabetic wounds in vivo. Sub-aim 1: Determine the efficacy of tRES+HESP on wound healing in a newly developed diabetic chronic wound model in db/db mice. Sub-aim 2: Determine the role of ANGPT1 in the tRES+HESP-induced improvement in wound healing in vivo. Sub-aim 3: Discover new proteins and pathways responsible for the benefit of tRES+HESP treatment in diabetic wound repair in vivo using state- of-the-art proteomics. The outcome of the proposed research will determine the efficacy of topical application of this formula, tRES+HESP, in diabetic wound healing, and will unveil underlying molecular mechanisms for its beneficial effect. Since tRES+HESP has not been approved by the FDA to treat diabetic wound healing yet, these results may facilitate the FDA approval of this coformulation in diabetic wound treatment.