Xu Shufang, Zhang Heng, Zou Hui, Chen Zhitao, Jiang Ting, Huang Mengjun
Key Laboratory for Molecular Diagnosis of Hubei, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Department of Clinical Nutrition, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430014, Hubei, China.
Appl Biochem Biotechnol. 2024 Nov;196(11):7703-7721. doi: 10.1007/s12010-024-04935-1. Epub 2024 Mar 28.
Diabetes is a major global health concern. This study aimed to investigate the correlation between differentially expressed lncRNAs in mice with type 2 diabetes mellitus (T2DM) and alterations in the intestinal flora and intestinal pathology. A T2DM mouse model was constructed by feeding mice a high-fat diet. Serum fat metabolism-related indices and insulin levels were biochemically detected. Serum inflammatory factors (IL-1β, IL-6, TNF-α, IL-10) and endotoxin (LPS) were measured by ELISA. Histopathological changes in the small intestines of mice were observed by HE. The short-chain fatty acid (SCFA) content was analyzed using GC-MS. Analysis of altered intestinal flora in T2DM mice was performed using a 16sRNA sequencing assay. Differences in lncRNA expression profiles in small intestinal tissues were analyzed using RNA-seq assays. Spearman's correlation analysis was used to correlate the expression of candidate lncRNAs with changes in differential gut flora. Spearman's correlation analysis was used to analyze the correlation between the expression of candidate differentially expressed lncRNAs, small intestinal permeability, and glucose absorption. We found that serum levels of LPS, BUN, Scr, TC, TG, LDL-C, IL-1β, IL-6, and TNF-α were elevated and levels of HDL-C, insulin, and IL-10 were decreased in T2DM mice. The ileal enterochromes of T2DM mice were disorganized and broken, the number of enterochromes was reduced, the local epithelial cells were necrotic, and the plasma membrane layer was locally absent. In addition, the protein expression of ZO-1 and occludin was decreased, and the protein expression of SGLT-1 and GLUT-2 was elevated in the model group compared to the control group. The levels of Acetic acid, Propionic acid and Butyric acid were decreased and the levels of Isobutyric acid and Isovaleric acid were increased, the abundance of beneficial bacteria was decreased and the abundance of harmful bacteria was increased in the feces of T2DM mice. RNA-seq identified nine differentially expressed lncRNAs (LINC00675, Gm33838, Gm11655, LOC6613926, LOC6613788, LOC6613791, LOC6613795, Arhgap27os3, and A330023F24Rik). In addition, we found significant correlations between differentially expressed lncRNAs and a variety of intestinal flora, as well as between small intestinal permeability and glucose absorption. A significant correlation was observed between differentially expressed lncRNAs in the intestinal tissues of T2DM mice and intestinal flora imbalance, small intestinal permeability, and glucose absorption.
糖尿病是全球主要的健康问题。本研究旨在探讨2型糖尿病（T2DM）小鼠中差异表达的长链非编码RNA（lncRNA）与肠道菌群改变及肠道病理变化之间的相关性。通过给小鼠喂食高脂饮食构建T2DM小鼠模型。生化检测血清脂肪代谢相关指标和胰岛素水平。采用酶联免疫吸附测定法（ELISA）检测血清炎症因子（IL-1β、IL-6、TNF-α、IL-10）和内毒素（LPS）。通过苏木精-伊红（HE）染色观察小鼠小肠的组织病理学变化。使用气相色谱-质谱联用仪（GC-MS）分析短链脂肪酸（SCFA）含量。采用16sRNA测序分析法对T2DM小鼠肠道菌群的改变进行分析。使用RNA测序分析法分析小肠组织中lncRNA表达谱的差异。采用Spearman相关性分析将候选lncRNA的表达与肠道菌群差异变化进行关联。采用Spearman相关性分析分析候选差异表达lncRNA的表达、小肠通透性和葡萄糖吸收之间的相关性。我们发现，T2DM小鼠血清中LPS、尿素氮（BUN）、肌酐（Scr）、总胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白胆固醇（LDL-C）、IL-1β、IL-6和TNF-α水平升高，高密度脂蛋白胆固醇（HDL-C）、胰岛素和IL-10水平降低。T2DM小鼠回肠肠嗜铬细胞排列紊乱且破碎，肠嗜铬细胞数量减少，局部上皮细胞坏死，局部质膜层缺失。此外，与对照组相比，模型组中紧密连接蛋白1（ZO-1）和闭合蛋白的蛋白表达降低，而钠-葡萄糖协同转运蛋白1（SGLT-1）和葡萄糖转运蛋白2（GLUT-2）的蛋白表达升高。T2DM小鼠粪便中乙酸、丙酸和丁酸水平降低，异丁酸和异戊酸水平升高，有益菌丰度降低，有害菌丰度增加。RNA测序鉴定出9种差异表达的lncRNA（LINC00675、Gm33838、Gm11655、LOC6613926、LOC6613788、LOC6613791、LOC6613795、Arhgap27os3和A330023F24Rik）。此外，我们发现差异表达的lncRNA与多种肠道菌群之间，以及小肠通透性和葡萄糖吸收之间存在显著相关性。T2DM小鼠肠道组织中差异表达的lncRNA与肠道菌群失衡、小肠通透性和葡萄糖吸收之间存在显著相关性。
