a Department of Medical College, Dalian University , Liaoning , China.
b Beijing International Travel Health Care Center of Beijing Entry-Exit Inspection and Quarantine Bureau , Beijing , China.
Pharm Biol. 2019 Dec;57(1):238-244. doi: 10.1080/13880209.2019.1588905.
Osthole is a natural coumarin compound most frequently extracted from plants of the Apiaceae family such as Cnidium monnieri (L.) Cusson, Angelica pubescens Maxin.f., and Peucedanum ostruthium (L.). Osthole is considered to have potential therapeutic applications for the treatment of diseases including epilepsy. However, the mechanism of osthole induced-apoptosis in BV-2 microglia cells is not yet clear.
To investigate the molecular mechanisms underlying the effect of osthole on PI3K/AKt/mTOR expression in kainic acid (KA)-activated BV-2 microglia cells.
Optimal culture concentration and time of osthole were investigated by MTT assay. The concentration of osthole was tested from 10 to 400 μM and the culture time was tested from 2 to 72 h. Ultrastructure difference among control, KA and osthole group was analyzed under transmission electron microscope. The mRNA expression of PI3K/AKt/mTOR was investigated using reverse transcription (RT)-PCR and the protein expression was investigated using western blotting and immunofluorescence assay. Apoptosis rate of BV-2 cells between each group was measured by flow cytometry.
IC for cell viability of BV-2 cells by osthole was 157.7 µM. Treated with osthole (140 µM) for 24 h significantly increased the inhibition rate. Pretreatment with osthole inhibited the KA-induced PI3K/AKt/mTOR mRNA and protein expression. The results of flow cytometry analysis showed that the apoptotic rate of osthole group was obviously higher than KA group.
Date showed that osthole may be useful in the treatment of epilepsy and other neurodegenerative diseases that are characterized by over expression of PI3K/Akt/mTOR.
蛇床子素是一种天然香豆素化合物，主要从伞形科植物中提取，如蛇床子、独活和前胡。蛇床子素被认为具有治疗癫痫等疾病的潜在治疗应用。然而，蛇床子素诱导 BV-2 小胶质细胞凋亡的机制尚不清楚。
研究蛇床子素对红藻氨酸（KA）激活的 BV-2 小胶质细胞中 PI3K/AKt/mTOR 表达的影响及其分子机制。
通过 MTT 法研究蛇床子素的最佳培养浓度和时间。测试蛇床子素的浓度范围为 10-400μM，培养时间为 2-72h。用透射电镜分析对照组、KA 组和蛇床子素组之间的超微结构差异。采用逆转录（RT）-PCR 检测 PI3K/AKt/mTOR 的 mRNA 表达，用 Western blot 和免疫荧光法检测蛋白表达。用流式细胞术测量各组 BV-2 细胞的凋亡率。
BV-2 细胞的 IC50 为 157.7μM。用 140μM 的蛇床子素处理 24h 后，抑制率显著增加。蛇床子素预处理抑制了 KA 诱导的 PI3K/AKt/mTOR mRNA 和蛋白表达。流式细胞术分析结果显示，蛇床子素组的凋亡率明显高于 KA 组。
数据表明，蛇床子素可能对癫痫和其他以 PI3K/Akt/mTOR 过度表达为特征的神经退行性疾病的治疗有一定作用。
