为探究植物乳植杆菌N1发酵对黑木耳多糖的影响,采用乙醇沉淀提取多糖,经Sevag法脱蛋白,DEAE-Sepharose Fast Flow和Sephacryl S-300柱分离纯化,得到了纯化的黑木耳发酵多糖FAAP-2-1(fermented Auricularia auricula-judae polysaccharide-2-1,FAAP-2-1),建立胰岛素抵抗HepG2(insulin resistance HepG2, IR-HepG2)细胞模型与斑马鱼糖尿病模型进行降血糖活性进行研究,利用实时荧光定量PCR法对其降血糖机制进行研究。结果发现,FAAP-2-1的相对分子质量为1.48×106 Da,主要由半乳糖、甘露糖、木糖、岩藻糖、葡萄糖、阿拉伯糖、半乳糖醛酸、葡萄糖醛酸8种单糖组成,其比例为4.27∶55.27∶0.36∶1.90∶16.34∶1.54∶2.51∶17.81。细胞实验和动物实验表明,FAAP-2-1可以恢复IR-HepG2细胞的葡萄糖消耗量,使糖尿病斑马鱼血糖下降73.52%,上调糖尿病相关基因AKT、PI3K、IRS和GLUT2的相对表达量,通过PI3K/AKT信号通路恢复和改善糖尿病斑马鱼葡萄糖代谢异常的症状,改善胰岛素抵抗。
To determine the impact of Lactiplantibacillus plantarun N1 fermentation on Auricularia auricula-judae polysaccharides, the polysaccharides were extracted using ethanol precipitation, deproteinized via the Sevag method, and further purified using DEAE-Sepharose Fast Flow and Sephacryl S-300 columns, resulting in the refined polysaccharide designated as fermented Auricularia auricula-judae polysaccharide-2-1 (FAAP-2-1). The hypoglycemic activities of FAAP-2-1 were evaluated using insulin-resistant HepG2(IR-HepG2) cell models and diabetic zebrafish models.Real-time qPCR was utilized to investigate the mechanisms underlying glucose reduction.Results showed that FAAP-2-1, with a molecular weight of 1.48×106 Da, primarily comprises eight monosaccharides:galactose, mannose, xylose, fucose, glucose, arabinose, galacturonic acid, and glucuronic acid, at ratios of 4.27∶55.27∶0.36∶1.90∶16.34∶1.54∶2.51∶17.81.Experiments indicated that FAAP-2-1 restores glucose consumption in IR-HepG2 cells and decreases blood glucose in diabetic zebrafish by 73.52%.Furthermore, FAAP-2-1 upregulates the expression of genes related to diabetes, including AKT, PI3K, IRS, and GLUT2, thereby ameliorating glucose metabolism abnormalities in diabetic zebrafish and improving insulin sensitivity via the PI3K/AKT pathway.
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