Ginsenoside Rg1调控异体ASCs旁分泌功能促进糖尿病创面的修复及分子机制

Diabetic Ulcers (DU) is one of the commonly most complications,which it is high incidence, longer treatment and harder healing. It has become the research focus, but the mechanism is unclear. Adipose-derived stem cells(ASCs) can promote repair and regeneration of the injured tissue, while the research about ASCs in DU wounds healing is not deeply opened.Our previous studies have confirmed that the ginsenoside Rg1 (Rg1) can up-regulate the paracrine function and HIF-1 alpha expression of ASCs, then subsequently to enhance the downstream related genes exprseeion and realize its effect on promoting tissue regeneration. These data suggested that Rg1 very likely play an important role in promoting tissue repair and regeneration by HIF-1α regulating the paratine of ASCs. On the basis of our previous research works, we hypothesized that Rg1 could be regulated HIF-1α expression of ASCs to achieve DU wounds healing, which is likely to be its key gene in miRNA31/HIF-1α and PI3K/Akt singaling pathway. We designed a series of experiments to detect the expression of downstream target genes in vivo and in vitro, including constructed wildtype and mutant dual luciferase reporter vectors of FIH-1-3’UTR, miRNA31 expression vector.we verified miRNA31/HIF-1α and then inhibited or activated PI3K/Akt, small hairpin RNA (shRNA) was used to silence the HIF-1α and AKT genes in ASCs. Our aim is illuminated that the molecular mechanisms and efficacy of how to promote wounds healing of DU by using the ASCs with Rg1 pretreament. Finally, we hope that our studies could provide a new approach and theoretical support for clinical application for ASCs widely applying to clinical therapy in regeneration medicine.

糖尿病溃疡（DU）发生率高、难愈合，是晚期糖尿病常见并发症之一，也是当前研究热点，但相关机制尚不明确。研究表明脂肪干细胞（ASCs）能促组织再生修复，但在促DU愈合研究方面欠深入。我们前期证实，人参皂苷Rg1（Rg1）能上调HIF-1α表达、增强ASCs旁分泌及下游相关因子表达促组织再生。提示Rg1可能通过HIF-1α调控ASCs旁分泌在促组织再生中发挥重要作用。本项目拟在此基础上提出Rg1调控ASCs 中miRNA31/HIF-1α、PI3K/Akt通路关键基因HIF-1α表达以促DU愈合的设想，构建FIH-1-3’UTR野生/突变型双荧光素酶及miRNA31表达载体，验证miRNA31/HIF-1α，激活/抑制PI3K/Akt， shRNA沉默ASCs中HIF-1α、AKT，检测下游靶基因，体内外实验阐明Rg1调控ASCs旁分泌促DU愈合的机制，为ASCs治疗DU提供新路径及理论依据。