■本研究旨在通过对糖尿病和糖耐量受损的多组学分析,探讨高血糖及其相关血管病变的发病机制。并使用细胞实验验证机制。
■在这项研究中,我们对糖尿病的宏基因组测序数据进行了综合分析,以探索与糖尿病发生相关的关键属。随后,被诊断为糖耐量受损(IGT)的参与者,和健康的受试者,被招募用于粪便和血液样本收集。使用16SrDNA测序和液相色谱质谱分析了肠道微生物群(GM)及其相关代谢产物的生态失调,分别。通过mRNA测序和数据独立获取技术评估基因和蛋白质表达的调节,分别。利用实时荧光定量PCR研究了GM菌群失调影响高血糖及其相关血管病变的具体机制,西方印迹,和酶联免疫吸附测定技术在HepG2细胞和中性粒细胞中的应用。
■根据公布的数据,与糖尿病相关的转基因中的关键可变属被确定为布劳蒂亚,乳酸菌,拟杆菌,普雷沃氏菌,粪杆菌,双歧杆菌,Ruminococus,梭菌属,和衣原体.相关的代谢途径被鉴定为胆酸盐降解和L-组氨酸生物合成。值得注意的是,与在糖尿病患者中观察到的那些相比,在IGT患者中Blautia和Faecalibacterium表现出相似的改变。和转基因代谢物,牛磺熊去氧胆酸(TUDCA)和肌肽(CARN,组氨酸和丙氨酸的下游代谢物)都被发现减少,进而调节血浆中蛋白质和中性粒细胞中mRNA的表达。随后的实验集中在胰岛素样生长因子结合蛋白3和白细胞介素6上,因为它们对血糖调节和相关的血管炎症有影响。发现两种蛋白质在HepG2细胞和嗜中性粒细胞中被TUDCA和CARN抑制。
■从IGT到糖尿病的整个过程中,发生了GM的菌群失调,特征是布劳特氏菌的增加和粪杆菌的减少,导致TUDCA和CARN水平降低,减轻了它们对胰岛素样生长因子结合蛋白3和白介素6表达的抑制作用,从而促进了高血糖症和相关血管病变的发展。
UNASSIGNED: This study aims to investigate the pathogenesis of hyperglycemia and its associated vasculopathy using multiomics analyses in diabetes and impaired glucose tolerance, and validate the mechanism using the cell experiments.
UNASSIGNED: In this study, we conducted a comprehensive analysis of the metagenomic sequencing data of diabetes to explore the key genera related to its occurrence. Subsequently, participants diagnosed with impaired glucose tolerance (IGT), and healthy subjects, were recruited for fecal and blood sample collection. The dysbiosis of the gut microbiota (GM) and its associated metabolites were analyzed using 16S rDNA sequencing and liquid chromatograph mass spectrometry, respectively. The regulation of gene and protein expression was evaluated through mRNA sequencing and data-independent acquisition technology, respectively. The specific mechanism by which GM dysbiosis affects hyperglycemia and its related vasculopathy was investigated using real-time qPCR, Western blotting, and enzyme-linked immunosorbent assay techniques in HepG2 cells and neutrophils.
UNASSIGNED: Based on the published data, the key alterable genera in the GM associated with diabetes were identified as Blautia, Lactobacillus, Bacteroides, Prevotella, Faecalibacterium, Bifidobacterium, Ruminococcus, Clostridium, and Lachnoclostridium. The related metabolic pathways were identified as cholate degradation and L-histidine biosynthesis. Noteworthy, Blautia and Faecalibacterium displayed similar alterations in patients with IGT compared to those observed in patients with diabetes, and the GM metabolites, tauroursodeoxycholic acid (TUDCA) and carnosine (CARN, a downstream metabolite of histidine and alanine) were both found to be decreased, which in turn regulated the expression of proteins in plasma and mRNAs in neutrophils. Subsequent experiments focused on insulin-like growth factor-binding protein 3 and interleukin-6 due to their impact on blood glucose regulation and associated vascular inflammation. Both proteins were found to be suppressed by TUDCA and CARN in HepG2 cells and neutrophils.
UNASSIGNED: Dysbiosis of the GM occurred throughout the entire progression from IGT to diabetes, characterized by an increase in Blautia and a decrease in Faecalibacterium, leading to reduced levels of TUDCA and CARN, which alleviated their inhibition on the expression of insulin-like growth factor-binding protein 3 and interleukin-6, contributing to the development of hyperglycemia and associated vasculopathy.