Ski募集HDAC1调节FOXO1乙酰化水平促进糖尿病难愈创面愈合的分子机制研究

Diabetes wound healing is a difficult problem in clinical. The research showed that the cell proliferation will reverse the apoptosis which induced by FOXO1 under hyperglycemia condition is the key to dalay wound healing. We confirmed that Ski can promote wound healing of DM, but its mechanism is not clear. The literature indicate Ski, as a corepressor, can recruit HDAC1 and mediate the deacetylation effect of its nuclear factor. The pre-experimen confirmed the combination of Ski and FOXO1 can significantly reduce the acetylation level of FOXO1. Therefore, it is speculated that Ski combined with FOXO1 and raised deacetylation of FOXO1 and then reverse promoting the apoptosis of cells to inducing the proliferation may be an important mechanism of Ski promoting DM wound healing. In this study, in hyperglycemia-induced-cell injury model, we comfirm the interaction of Ski、HDAC1 and FOXO1 using IP firstly, and then detect.level of FOXO1 acetylation and the ability of cell apoptosis and proliferation by WB, Tunel, BrdU and other methods. Finally, we use HDAC1 inhibitors or siRNA to explore the pro-healing of Ski was achieved by regulating the acetylation level of FOXO1, This study will provide a solid foundation for development of the new pro-healing drugs based on Ski

糖尿病（dibetes mellitus，DM）创面难愈是临床棘手问题，研究表明高糖诱导核因子FOXO1乙酰化将其促创面愈合逆转成延迟创面愈合是导致创面难愈的关键，我们已证实Ski具有促DM创面愈合的作用，但机制还不清楚，文献表明Ski可介导HDAC1去乙酰化调节与其结合的核因子，预实验证实Ski与FOXO1的结合可降低FOXO1的乙酰化水平，因而推测Ski结合FOXO1并募集HDAC1去乙酰化FOXO1可能是Ski促DM创面愈合的重要机制。本研究拟在高糖诱导的细胞损伤模型中，首先采用IP验证Ski、FOXO1和HDAC1的相互作用，进而采用WB、TUNEL、BrdU等法检测Ski作用后FOXO1的乙酰化水平及细胞凋亡、增殖及胞内ROS生成情况，最后采用HDAC1激动剂或siRNA证实Ski是通过HDAC1去乙酰化FOXO1来促进DM创面愈合的，为将Ski作为促愈药物的研发提供理论依据。

成纤维细胞增殖受抑和凋亡增加为糖尿病患者伤口难愈的重要原因，也是促愈治疗的重要靶点，已知Ski具有促进成纤维细胞增殖、抑制凋亡等作用，但其对高糖环境下的成纤维细胞有何作用还不清楚。本研究采用大鼠原代成纤维细胞，在明确高糖导致细胞增殖受抑、凋亡升高的基础上，发现高糖可导致Ski蛋白水平下降；相反的，提高Ski水平可显著改善高糖诱导的细胞增殖受抑和促凋亡作用，且其作用不影响Smad2/3磷酸化水平, 进一步发现提高Ski水平可增加其与FoxO1结合进而降低FoxO1的乙酰化水平，且干扰FoxO1后可导致提高Ski水平显著改善高糖诱导的细胞增殖受抑和促凋亡的作用丧失, 并且提高Ski水平可通过HDAC1介导去乙酰化作用而降低FoxO1的乙酰化水平。这些结果不仅证实Ski是一个在高糖下可调节成纤维细胞增殖和抑制其凋亡作用的分子，而且发现Ski可调节FoxO1信号途径，为通过调控Ski而达到直接干预成纤维细胞增殖、凋亡来实现促进糖尿病难愈伤口愈合提供了实验依据。