银杏内酯B对高胆固醇诱导的PC12细胞胆固醇含量的影响

Effect of ginkgolide B on cholesterol content in PC12 cells under high cholesterol culture

目的:基于体外血脑屏障环境下观察银杏内酯B(ginkgolide B,GB)对高胆固醇诱导PC12细胞胆固醇含量的影响。方法:利用Transwell共培养脑微血管内皮细胞和星形胶质细胞建立体外血脑屏障,并与PC12细胞共培养。在Transwell内皮细胞侧添加40μmol/L胆固醇,并选择不同浓度银杏内酯B分别干预上述模型。观察PC12细胞的形态学变化,通过MTT法筛选银杏内酯B的干预浓度,用HPLC法检测PC12细胞内胆固醇含量的变化。结果:倒置相差显微镜观察发现PC12细胞大多呈圆形、短梭形或三角形,胞浆伸展,细胞两极有短突起,细胞折光性好,胞核可见。通过MTT确定的银杏内酯B干预剂量分别为低剂量1μmol/L、中剂量5μmol/L和高剂量25μmol/L。HPLC法检测显示模型组PC12细胞内胆固醇含量升高,与对照组相比差异显著(P<0.01)。银杏内酯B干预上述模型16 h后,25μmol/L GB降胆固醇的效果最佳,与模型组相比差异显著(P<0.05),25μmol/L GB与阳性对照药辛伐他汀相比差异无显著(P>0.05)。结论:中药银杏叶有效单体GB可以降低体外血脑屏障环境下PC12细胞内胆固醇含量。

AIM： To explore the effect of ginkgolide B（GB） on high cholesterol-induced dysfunction in PC12 cells.METHODS： The rat brain microvascular endothelial cells and astrocytes were used to establish the blood-brain barrier（BBB） model by in vitro co-culture in the Transwell device with PC12 cells.Cholesterol at the concentration of 40 μmol/L was added into the side of brain microvascular endothelial cells of the Transwell.Various concentrations of GB were applied to deal with the culture system.The morphological changes of the cells were observed under microscope.The safe doses of GB were screened by MTT assay.The level of cholesterol in the PC12 cells was determined by the high-performance liquid chromatography.RESULTS： The morphology of PC12 cells in control group was normal,with round,short spindle or triangle shape,cytoplasm extension,two-pole short processes,good refraction and obvious nucleus.The doses of GB determined by MTT assay were 1 μmol/L for low dose,5 μmol/L for middle dose and 25 μmol/L for high dose.Compared with control group,the cholesterol obviously increased in model group with statistical difference（P0.05）.GB at dose of 25 μmol/L for 16 h exposure had the optimum results to decrease the cholesterol level in PC12 cells.However,no significant difference between GB high dose group and simvastatin group was observed（P0.05）.CONCLUSION： The active monomer of ginkgo leaf,GB,may effectively reduce the cholesterol content in PC12 cells with the condition of BBB in vitro.