【摘要】 目的：本研究基于生物信息学分析筛选2型糖尿病（Type 2 Diabetes Mellitus, T2DM）与铁死亡之间的新通路，并通过细胞实验进行验证。方法：（1）从Gene Expression Omnibus(GEO)数据库下载T2DM数据集GSE25724，并从铁死亡数据库Ferr Db中获取铁死亡基因，寻找T2DM与铁死亡的共同差异基因，并进行富集分析；（2）体外实验：选择小鼠胰岛瘤MIN6细胞，通过25 mmol/L高糖联合200μmol/L棕榈酸钠（High Glucose and High Sodium Palmitate, GP）干预，建立T2DM胰岛β细胞损伤模型；通过Western Blot检测蛋白表达和荧光实验检测细胞活性氧和线粒体膜电位变化。结果：T2DM数据集与铁死亡共有38个差异基因，其中13个上调，25个下调；富集分析结果显示这些差异基因主要富集于自噬及线粒体自噬；Western Blot结果显示，与对照组相比，PINK1/Parkin蛋白表达下降，LC3Ⅱ和P62蛋白表达上升（P<0.05）；荧光结果显示线粒体与溶酶体的荧光共定位黄色荧光下降，线粒体膜电位水平下降，ROS水平上升。结论：线粒体自噬通路蛋白PINK1/Parkin在2型糖尿病铁死亡的进程中下调；线粒体自噬可能是减少胰岛β细胞铁死亡的一个重要保护机制。更多还原

【Abstract】 Objective: To identify novel pathways linking ferroptosis and type 2 diabetes mellitus(T2DM) based on bioinformatics analysis and to validate the findings through cellular experiments. Methods: (1)The T2DM dataset GSE25724 was downloaded from the Gene Expression Omnibus(GEO) database, and ferroptosis-related genes were obtained from the Ferr Db database. Common differentially expressed genes between T2DM and ferroptosis were identified and subjected to enrichment analysis.(2)In vitro experiments were conducted using mouse insulinoma MIN6 cells. A T2DM-related pancreatic β-cell injury model was established by treatment with 25 mmol/L high glucose combined with 200 μmol/L sodium palmitate(high glucose and high palmitate, GP). Protein expression was detected by Western blotting, and changes in reactive oxygen species(ROS) levels and mitochondrial membrane potential were assessed using fluorescence assays. Results: A total of 38 common differentially expressed genes were identified between the T2DM dataset and ferroptosis, including 13 upregulated and 25 downregulated genes. Enrichment analysis indicated that these genes were mainly involved in autophagy and mitophagy pathways. Western blot results showed decreased expression of PINK1 and Parkin proteins, along with increased expression of LC3Ⅱ and P62 proteins in the GP group compared with the control group(P<0.05). Fluorescence assays demonstrated reduced mitochondrial – lysosomal colocalization, decreased mitochondrial membrane potential, and increased ROS levels.Conclusion: The mitophagy-related proteins PINK1 and Parkin are downregulated during ferroptosis in T2DM.Mitophagy may represent an important protective mechanism against ferroptosis in pancreatic β-cells.更多还原