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目的:考察灯盏细辛中咖啡酸及其衍生物对体外培养大鼠大脑皮层神经细胞存活的影响。方法:取出生1d内的乳鼠大脑皮层制成细胞悬液,接种于经多聚赖氨酸包被的96孔板中。于培养6d后,分别加入培养液及不同浓度的咖啡酸类药物,继续培养1d,采用四甲基偶氮唑盐(MTT)比色法测量其存活细胞的吸收值,同时行神经特异性烯醇化酶(NSE)检查。结果:NSE检查结果表明,培养6d的存活细胞大部分均为神经元。与空白对照组比较,咖啡酸(100,400mg·L-1)、咖啡酸甲酯(4,20mg·L-1)、咖啡酸乙酯(20mg·L-1)、3,4-二乙酰基咖啡酸(4,20,100mg·L-1)的存活细胞数均明显增加(P<0.05,P<0.01)。结论:各有效成分均能不同程度促进体外培养的大脑皮层神经细胞存活,且以3,4-二乙酰基咖啡酸活性较强。
OBJECTIVE: To investigate the effect of caffeic acid and its derivatives from Erigeron Breviscapus on the survival of cultured rat cerebral cortical neurons. Methods: The neonatal rat cerebral cortex was taken out to prepare cell suspension and inoculated into poly-lysine-coated 96-well plates. After cultured for 6 days, the culture medium and different concentrations of caffeic acid were added respectively and cultured for 1 day. MTT assay was used to measure the absorbance of surviving cells. At the same time, nerve-specific enamel Alcoholase (NSE) examination. Results: The results of NSE showed that most of the surviving cells cultured for 6 days were neurons. Compared with the blank control group, Caffeic acid (100,400 mg · L -1), Caffeic acid methyl ester (4,20 mg · L -1), Caffeic acid ethyl ester (20 mg · L -1), 3,4-diacetyl Caffeic acid (4, 20 and 100 mg · L -1) significantly increased the number of viable cells (P <0.05, P <0.01). CONCLUSION: All the active ingredients can promote the survival of cultured neural cortex neurons in vitro to a great extent, and 3,4-diacetyl caffeic acid is more active.