Mechanistic study of Small-molecular Therapy in diabetic Wound Healing

糖尿病足溃疡导致截肢是一个主要的健康问题，在3000万糖尿病患者中约有20%受到影响 在美国的病人。目前的方案效果有限，截肢率仍然很高。因此， 了解具有翻译潜力的化合物的分子机制是制造 伤口护理方案的突破。内皮细胞(ECs)是人体不可缺少的细胞成分 伤口血管生成。然而，糖尿病患者的EC功能受损。两者的共同提法 膳食化合物-反式白藜芦醇(Tres)和橙皮素(Hesp)-改善葡萄糖代谢状况和 超重和肥胖者通过诱导乙二酸酶1(GLO1)基因表达来实现动脉功能 一种在糖酵解过程中解毒活性代谢物并保护细胞免受糖基化压力的酶。我们的 试验数据表明，TRES+HESP通过增加GLO1促进糖尿病动物的伤口愈合 表情。然而，它的作用可能远远不止诱导GLO1的表达，因为tres+hesp处理过 内皮细胞产生许多促血管生成因子，包括血管生成素-1(ANGPT1)，它在 血管生成。因此，确定在血管生成过程中受tres+hesp调控的蛋白质是至关重要的。 和组织修复。这个项目的目标是填补关于tres+hesp在救援中的作用的知识空白。 糖尿病血管生成受阻，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促进血管生成并促进糖尿病创面愈合 通过增强GLO1和一种强有力的促血管生成因子ANGPT1的表达，以及通过新的 糖尿病伤口修复的关键途径中额外蛋白质的变化。目标1：分子鉴定 Tres+Hesp体外诱导人真皮微血管内皮细胞的通路和蛋白质变化。子目标 1：确定TRES+HESP在多大程度上能在体外挽救糖尿病内皮细胞功能。次级目标2： 确定ANGPT1在TRES+Hesp诱导的体外血管生成中有多重要。次级目标3：发现新的蛋白质 以及TRES+HESP治疗对内皮细胞功能的影响的途径。 最先进的蛋白质组学。目的2：确定TRES+HESP及其潜在分子的治疗潜力 体内慢性糖尿病创面的机制。次级目标1：确定TRES+Hesp对伤口的疗效 Db/db小鼠新发展的糖尿病慢性创面模型的愈合。次级目标2：确定 TRES+HESP中的ANGPT1在体内可促进创面愈合。次级目标3：发现新的蛋白质 以及在体内糖尿病创面修复中TRES+Hesp治疗受益的途径。 最先进的蛋白质组学。拟议的研究结果将决定外用药物的疗效。 Tres+Hesp在糖尿病伤口愈合中的作用，并将揭示其潜在的分子机制 有益的效果。由于TRES+HESP尚未被FDA批准用于治疗糖尿病伤口愈合， 这些结果可能有助于FDA批准该联合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.