Department of Neurology, Beijing Anzhen Hospital, Capital Medical University, Beijing, 100029, China.
Research & Development Centre of Chengdu Baiyu Pharmaceutical Co., Ltd, Chengdu, 610041, China.
Appl Biochem Biotechnol. 2022 Jun;194(6):2448-2464. doi: 10.1007/s12010-022-03828-5. Epub 2022 Feb 7.
NF-κB signaling has been reported to play a key regulatory role in the pathogenesis of Alzheimer's disease (AD). The purpose of this study is to investigate the effects of ginkgolide on cell viability in an AD cellular model involving an APP/PS1 double gene-transfected HEK293 cell line (APP/PS1-HEK293) and further explore the mechanisms of action related to NF-κB signaling. The optimal time point and concentration of ginkgolide for cell proliferation were screened using a cell counting kit-8 assay. Based on the results, an in vitro study was performed by co-culture of APP/PS1-HEK293 with different dosages of ginkgolide, followed by an enzyme-linked immunosorbent assay to measure the levels of supernatant tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6, as well as western blotting and real-time polymerase chain reaction to detect intracellular protein and mRNA expression of NF-κB p65, IκBa, Bcl-2, and Bax. APP/PS1-HEK293 cells exhibited the highest cell viability at a concentration of 100 µg/ml after 48 h of treatment with ginkgolide. The supernatant levels of TNF-α, IL-1β, and IL-6 in the high-dosage ginkgolide-treated groups were lower than those in the control group. Compared with the control group, there were decreased intracellular protein and mRNA expression of NF-κB p65 and Bax, but increased protein and mRNA expression of IκBa in both high-dosage and low-dosage groups. Ginkgolide may enhance cell viability, indicative of its neuroprotective effects on AD, at least partially via suppression of the NF-κB signaling pathway involving anti-apoptosis and anti-inflammation mechanisms. Therefore, ginkgolide might be a promising therapeutic agent against AD.
NF-κB 信号通路在阿尔茨海默病（AD）的发病机制中发挥着关键的调节作用。本研究旨在探讨银杏内酯对 APP/PS1 双基因转染的 HEK293 细胞系（APP/PS1-HEK293）AD 细胞模型中细胞活力的影响，并进一步探讨与 NF-κB 信号通路相关的作用机制。通过细胞计数试剂盒-8 检测筛选银杏内酯促进细胞增殖的最佳时间点和浓度。基于该结果，通过 APP/PS1-HEK293 与不同剂量银杏内酯的共培养进行体外研究，然后通过酶联免疫吸附试验测量上清液肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β 和 IL-6 的水平，以及通过 Western blot 和实时聚合酶链反应检测细胞内 NF-κB p65、IκBa、Bcl-2 和 Bax 的蛋白和 mRNA 表达。银杏内酯处理 48 小时后，浓度为 100μg/ml 时 APP/PS1-HEK293 细胞的活力最高。高剂量银杏内酯处理组的 TNF-α、IL-1β 和 IL-6 上清液水平低于对照组。与对照组相比，高剂量和低剂量组的 NF-κB p65 和 Bax 的细胞内蛋白和 mRNA 表达降低，而 IκBa 的蛋白和 mRNA 表达增加。银杏内酯可能通过抑制 NF-κB 信号通路（涉及抗凋亡和抗炎机制）增强细胞活力，对 AD 具有神经保护作用，至少部分如此。因此，银杏内酯可能是一种有前途的 AD 治疗药物。
