Ba Xiao-Hong, Min Lian-Qiu
Department of Neurology, the First Affiliated Hospital, Liaoning Medical University, Jinzhou 121001, China ; Key Laboratory of Brain and Spinal Cord Injury in Liaoning Province, Jinzhou 121001, China.
Indian J Pharmacol. 2015 Jan-Feb;47(1):101-4. doi: 10.4103/0253-7613.150372.
The aim was to observe the effects of the extract of Ginkgo biloba (EGb761) on the apoptosis of oxygen and glucose-deprived (OGD) human neuroblastoma cells (SH-SY5Y) cells and explore its mechanism.
SH-SY5Y cells were divided into normal control group, OGD group, OGD for 4 h and EGb761-pretreated groups including very low-concentration (20 μg/ml), low-concentration group (25 μg/ml), moderate-concentration group (50 μg/ml) and high-concentration group (100 μg/ml). Twenty four hours after reoxygenation, cell viability was determined with 3-[4, 5-dimehyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide assay, apoptosis rate was detected with annexin V-fluorescein isothiocyanate/propidium iodide double staining flow cytometry and the protein level of apoptosis-inducing factor (AIF) was observed with immunofluorescence technique in each group.
Cell viability was significantly lower in OGD group than in EGb761-pretreated groups, especially in moderate-concentration group (50 μg/ml) (P < 0.005). Apoptosis rate was significantly lower in EGb761-pretreated groups than in OGD group (P < 0.001). Immunofluorescent staining showed that there was AIF nuclear translocation in both EGb761-pretreated groups and OGD group, but AIF nuclear translocation was less in EGb761-pretreated groups than in OGD group.
EGb761 can reduce the apoptosis of OGD SH-SY5Y cells probably through inhibiting AIF nuclear translocation. This study provides a theoretical basis for the application of EGb761 in clinical practice.
观察银杏叶提取物(EGb761)对氧糖剥夺(OGD)的人神经母细胞瘤细胞(SH-SY5Y)凋亡的影响并探讨其机制。
将SH-SY5Y细胞分为正常对照组、OGD组、OGD 4小时组以及EGb761预处理组,EGb761预处理组包括极低浓度(20μg/ml)、低浓度组(25μg/ml)、中浓度组(50μg/ml)和高浓度组(100μg/ml)。复氧24小时后,采用3-[4,5-二甲基-2-噻唑基]-2,5-二苯基-2H-四氮唑溴盐法测定细胞活力,采用膜联蛋白V-异硫氰酸荧光素/碘化丙啶双染流式细胞术检测凋亡率,并用免疫荧光技术观察各组凋亡诱导因子(AIF)的蛋白水平。
OGD组细胞活力显著低于EGb761预处理组,尤其是中浓度组(50μg/ml)(P<0.005)。EGb761预处理组凋亡率显著低于OGD组(P<0.001)。免疫荧光染色显示,EGb761预处理组和OGD组均有AIF核转位,但EGb761预处理组的AIF核转位较OGD组少。
EGb761可能通过抑制AIF核转位来减少OGD的SH-SY5Y细胞凋亡。本研究为EGb761在临床实践中的应用提供了理论依据。