目的 探究露水草醋酸乙酯部位（ethyl acetate fraction of Cyanotis arachnoidea，ECA）对2型糖尿病（type 2 diabetes mellitus，T2DM）小鼠的改善作用及其机制。方法 通过高脂饲料喂养联合ip链脲佐菌素建立T2DM小鼠模型。ECA干预4周后，检测ECA对T2DM小鼠糖脂代谢生化指标及组织病理变化的影响。采用非靶向代谢组学分析各组小鼠血清中的代谢物变化及所涉及代谢通路。采用超高效液相色谱-四级杆飞行时间质谱表征ECA的化学组成，结合网络药理学方法筛选ECA改善T2DM的关键靶点及通路，并采用Western blotting验证相关靶标蛋白表达。结果 ECA可有效增加T2DM小鼠体质量并显著降低T2DM小鼠空腹血糖（P＜0.01），改善糖耐量、胰岛素抵抗及空腹胰岛素水平（P＜0.01）。ECA可显著改善T2DM小鼠血清中三酰甘油、总胆固醇、低密度脂蛋白胆固醇以及高密度脂蛋白胆固醇水平（P＜0.01），缓解肝脏和胰腺病理损伤。代谢组学分析显示，ECA主要调控18种差异代谢物介导的甘油磷脂代谢、嘌呤代谢以及赖氨酸降解等代谢通路。网络药理学结果显示，ECA可能通过调节蛋白激酶B1、磷脂酰肌醇4,5-二磷酸3-激酶催化亚基α、糖原合成酶激酶-3β（glycogen synthase kinase-3β，GSK-3β）等靶点介导的磷酸肌醇3-激酶（phosphoinositide 3-kinase，PI3K）/蛋白激酶B（protein kinase B，Akt）信号通路改善T2DM。Western blotting实验结果表明，与模型组比较，ECA显著上调小鼠肝脏组织中PI3K、Akt、GSK-3β的磷酸化水平（P＜0.05、0.01）。结论 ECA可能通过调控PI3K/Akt轴及恢复代谢稳态减轻T2DM小鼠的糖脂代谢异常。

Objective To explore the therapeutic effects and mechanisms of ethyl acetate fraction of Cyanotis arachnoidea (ECA) on type 2 diabetes mellitus (T2DM) in mice. Methods A mouse model of T2DM was established by administering a high-fat diet followed by intraperitoneal injection with streptozotocin. After ECA intervention for four weeks, the effects of ECA on serum biochemical indicators of glucolipid metabolism and histopathological changes in T2DM mice were evaluated. Untargeted metabolomics was employed to analyze metabolite alterations in the serum of mice from each group and to identify the involved metabolic pathways. The chemical constituents of ECA were characterized using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry. Integrated with network pharmacology, the key target groups responsible for the ameliorative effects of ECA on T2DM were screened. Furthermore, the expression levels of these key target proteins were validated by Western blotting confirmatory experiment. Results ECA effectively increased body weight and significantly reduced fasting blood glucose in T2DM mice (P < 0.01), and improved glucose tolerance, insulin resistance and fasting insulin levels (P < 0.01). Additionally, ECA significantly modulated serum lipid profiles, including triglycerides, total cholesterol, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol (P < 0.01), and alleviated pathological injuries in the liver and pancreas. Metabolomics analysis revealed that ECA primarily regulated metabolic pathways such as glycerophospholipid metabolism, purine metabolism and lysine degradation, which were mediated by 18 differential metabolites. Network pharmacology results indicated that ECA might ameliorate T2DM by targeting key molecules including protein kinase B1, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha, glycogen synthase kinase-3β (GSK-3β), and modulating the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway. Western blotting results demonstrated that ECA significantly up-regulated the phosphorylation levels of PI3K, Akt and GSK-3β in liver tissues of mice compared with model group (P < 0.05, 0.01). Conclusion ECA ameliorates glucolipid metabolism dysregulation in T2DM by modulating the PI3K/Akt axis and restoring metabolic homeostasis.

R285.5

四川省中医药管理局中医药科研专项（2024ZD026）；西南民族大学青藏高原研究科技创新团队（2024CXTD16）；四川省中医药管理局重大项目（民族药材系统研究与转化创新团队）（2023ZD05）