银杏内酯A、B对高胆固醇诱导的PC12细胞胆固醇代谢的影响

Effect of gingko lactone A and gingko lactone B on cholesterol metabolism in PC12 cells under high cholesterol culture

关键词:  银杏内酯A  银杏内酯B  胆固醇  PC12细胞  血脑屏障

Key words:  Ggingko lactone A  Gingko lactone B  Cholesterol  PC12 cells  blood-brain barrier

摘要:

[目的] 基于体外血脑屏障(BBB)环境, 观察高胆固醇环境下PC12细胞胆固醇的代谢情况及银杏内酯A、B对其的影响。[方法] 利用Transwell装置共培养星形胶质细胞和脑微血管内皮细胞建立体外BBB, 并与PC12细胞共培养。在Transwell 内皮细胞侧添加40 μmol/L胆固醇, 并分别给予30 μmol/L 银杏内酯A和25 μmol/L 银杏内酯B进行干预。干预结束后, 采用反转录聚合酶链式反应(RT-PCR)技术, 检测胆固醇代谢关键酶低密度脂蛋白受体相关蛋白-1(LRP-1)、胆固醇酰基转移酶(ACAT)、ATP结合盒转运子A1(ABCA1)及胆固醇24S-羟化酶基因(CYP46)在mRNA水平的表达。[结果] 高胆固醇环境下, LRP-1表达下调(P<0.01), ACAT 及ABCA1表达上调(P<0.05), CYP46的表达有上调趋势;药物干预后, 除LRP-1有不同程度的升高(P<0.05), 其余各指标均无明显变化。[结论] GKA、B对高胆固醇环境下PC12细胞胆固醇代谢关键酶LRP-1的表达有一定影响。

Abstract:

[Objective] To observe the cholesterol metabolism in PC12 cells with high cholesterol-induced dysfunction under in vitro blood-brain barrier (BBB)-based environment and to explore the effects of gingko lactone A and B(GKA and GKB). [Methods] The rat brain microvascular endothelial cells and astrocytes were used to establish the BBB model by co-cultured in the Transwell device with PC12 cells. Cholesterol, 40 μmol/L, was added into the Transwell endothelial cell side. GKA at a dose of 30 μmol/L and GKB at a dose of 25 μmol/L were applied to deal with the culture system respectively. After intervention, RT-PCR technique was used to detect the cholesterol metabolism key enzyme of LRP-1, ACAT, ABCA1 and CYP46. [Results] Under high cholesterol environment, the expression of LRP-1 decreased (P<0.01) and the expression of ACAT and ABCA1 increased (P<0.05). After intervention, the expression of LRP-1 was increased in varying degrees (P<0.05), but no significant changes were found in other enzymes. [Conclusion] GKA and GKB can affect the expression of LRP-1 in PC12 cells.