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目的观察毛蕊花苷(verbascoside)对过表达血红素氧化酶-1(heme oxygenase-1,HO-1)的大鼠原代星形胶质细胞铁离子沉积的影响,寻找抗阿尔兹海默病的新药物。方法体外培养和纯化大鼠原代星形胶质细胞,构建转染HO-1基因的星形胶质细胞模型,用CCK-8法检测细胞损伤,Western blotting法检测HO-1蛋白的表达,普鲁士蓝染色法检测细胞铁离子沉积。结果 95%以上的细胞呈胶质纤维酸性蛋白(GFAP)阳性,表明成功培养出星形胶质细胞;毛蕊花苷对HO-1基因转染后的原代星形胶质细胞具有不同程度的增殖抑制效应,抑制率与药物浓度呈正相关;转染HO-1基因入星形胶质细胞后,HO-1蛋白表达增加6.86倍;在施加毛蕊花苷干预后,20、40、60μmol·L~(-1)均能不同程度地降低HO-1蛋白的表达,同时不同程度地减少由过表达HO-1导致的铁离子沉积(20μmol·L~(-1)降低47.69%,40μmol·L~(-1)降低51.63%)。结论毛蕊花苷可以抑制过表达HO-1大鼠原代星形胶质细胞的铁离子沉积,可能成为针对“铁-氧化损伤”途径的抗阿尔兹海默病的新药物。
Objective To investigate the effect of verbascoside on iron deposition in rat primary astrocytes overexpressing heme oxygenase-1 (HO-1), and to find out the anti-Alzheimer’s New drug Methods Primary rat astrocytes were cultured and purified in vitro. The astrocyte model transfected with HO-1 gene was constructed. The cell injury was detected by CCK-8 assay. The expression of HO-1 protein was detected by Western blotting. Prussian blue staining assay of iron ion deposition. RESULTS: More than 95% of the cells were positive for glial fibrillary acidic protein (GFAP), indicating that astrocytes were successfully cultured. Verbascoside had different degrees of proliferation of primary astrocytes after HO-1 gene transfection The inhibitory effect was positively correlated with drug concentration. The HO-1 protein expression increased by 6.86-fold after transfection of HO-1 gene into astrocytes. After the intervention of verbascoside, -1) could reduce the expression of HO-1 protein to some extent, and at the same time, reduce the iron deposition induced by overexpression of HO-1 to a certain extent (20μmol·L -1, 47.69%, 40μmol·L -1 -1) by 51.63%). Conclusion Verbascoside can inhibit iron ion deposition in overexpressed primary astrocytes of HO-1 rats and may become a new anti-Alzheimer’s drug targeting the “iron-oxidative damage” pathway.