背景:2型糖尿病是一种内分泌紊乱,其特征是胰岛素敏感性受损,最终导致明显的疾病。脂肪干细胞(ASC)通过其免疫调节和分化能力在改善2型糖尿病及其并发症方面显示出有希望的潜力。然而,糖尿病微环境的高血糖可能对ASC的功能产生不利影响。在这里,我们调查了糖尿病环境中的ASC稳态和再生潜力。
方法:我们进行了数据收集和功能富集分析,以研究糖尿病微环境中MSCs的差异基因表达谱。接下来,将ASC在含有糖尿病血清(DS)或正常非糖尿病血清(NS)的培养基中培养6天和1个月的时间。进行了蛋白质组学分析,然后评估ASCs的凋亡,表面标记和DNA修复基因表达的变化,细胞内氧化应激,和差异化能力。ASC与糖尿病微环境之间的串扰由促炎和抗炎细胞因子和细胞因子受体的表达决定。
结果:糖尿病中MSCs差异表达基因的富集表明MSCs中氧化应激调节途径的改变。接下来,DS中ASCs的蛋白质组学分析揭示了与细胞凋亡增强相关的差异表达蛋白,DNA损伤和氧化应激,免疫调节和分化潜能改变。我们的实验证实了这些数据,并表明在DS中培养的ASCs遭受凋亡,细胞内氧化应激,和DNA修复缺陷.在糖尿病的情况下,ASCs还显示出成骨性受损,成脂,和血管生成分化能力。通过在DS中培养ASC,表明免疫调节潜力缺陷,可以显着改变促炎和抗炎细胞因子的表达。有趣的是,ASC显示诱导抗氧化应激基因和蛋白质,例如SIRT1,TERF1,Clusterin和PKM2。
结论:我们认为ASCs再生功能的这种恶化部分是由诱导的氧化应激和糖尿病炎症环境介导的。在ASC中诱导抗氧化应激因子可能表明对糖尿病微环境中增加的氧化应激的适应机制。
BACKGROUND: Type 2 diabetes is an endocrine disorder characterized by compromised insulin sensitivity that eventually leads to overt disease. Adipose stem cells (ASCs) showed promising potency in improving type 2 diabetes and its complications through their immunomodulatory and differentiation capabilities. However, the hyperglycaemia of the diabetic microenvironment may exert a detrimental effect on the functionality of ASCs. Herein, we investigate ASC homeostasis and regenerative potential in the diabetic milieu.
METHODS: We conducted data collection and functional enrichment analysis to investigate the differential gene expression profile of MSCs in the diabetic microenvironment. Next, ASCs were cultured in a medium containing diabetic serum (DS) or normal non-diabetic serum (NS) for six days and one-month periods. Proteomic analysis was carried out, and ASCs were then evaluated for apoptosis, changes in the expression of surface markers and DNA repair genes, intracellular oxidative stress, and differentiation capacity. The crosstalk between the ASCs and the diabetic microenvironment was determined by the expression of pro and anti-inflammatory cytokines and cytokine receptors.
RESULTS: The enrichment of MSCs differentially expressed genes in diabetes points to an alteration in oxidative stress regulating pathways in MSCs. Next, proteomic analysis of ASCs in DS revealed differentially expressed proteins that are related to enhanced cellular apoptosis, DNA damage and oxidative stress, altered immunomodulatory and differentiation potential. Our experiments confirmed these data and showed that ASCs cultured in DS suffered apoptosis, intracellular oxidative stress, and defective DNA repair. Under diabetic conditions, ASCs also showed compromised osteogenic, adipogenic, and angiogenic differentiation capacities. Both pro- and anti-inflammatory cytokine expression were significantly altered by culture of ASCs in DS denoting defective immunomodulatory potential. Interestingly, ASCs showed induction of antioxidative stress genes and proteins such as SIRT1, TERF1, Clusterin and PKM2.
CONCLUSIONS: We propose that this deterioration in the regenerative function of ASCs is partially mediated by the induced oxidative stress and the diabetic inflammatory milieu. The induction of antioxidative stress factors in ASCs may indicate an adaptation mechanism to the increased oxidative stress in the diabetic microenvironment.