Arslan Mehmet Enes, Türkez Hasan, Mardinoğlu Adil
Department of Molecular Biology and Genetics, Faculty of Science, Erzurum Technical University, Erzurum, Turkey.
Department of Medical Biology, Faculty of Medicine, Atatürk University, Erzurum, Turkey.
Int J Neurosci. 2021 Aug;131(8):745-754. doi: 10.1080/00207454.2020.1754211. Epub 2020 Apr 19.
To investigate neuroprotective properties of the farnesene sesquiterpene on the experimental Alzheimer's disease model .
Human neuroblastoma cell line (SHSY-5Y) was differentiated into neuron-like cells by using retinoic acid to constitute the Alzheimer's Disease model. β-amyloid 1-42 protein was applied to the transformed cells for 24 and 48 hours in a wide dose ranges (3.125-200 μM) to establish AD cytotoxicity. Then, farnesene was applied to cell cultures in a wide spectrum dose interval (1.625-100 μg/ml) to investigate neuroprotective effect against β-amyloid for 24 and 48 hours. 3-(4,5-dimethyl-thiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) release tests were executed to determine cytotoxicity in the Alzheimer model. Nuclear DNA integrity of cells was examined under the fluorescent microscope using the Hoechst 33258 staining method. Furthermore, acetylcholinesterase (AChE) activity, total antioxidant capacity (TAC) and total oxidative status (TOS) levels were analyzed to understand the protection mechanism of the farnesene application on the cell culture model. Finally, flow cytometry analysis was used to find out the cell death mechanism after beta-amyloid and farnesene application to the cell culture.
Cell viability tests revealed significant neuroprotection against β-amyloid toxicity in both 24 and 48 hours and the Hoechst 33258 fluorescence staining method showed a significant decrease in necrotic deaths after farnesene application in the cell cultures. Finally, flow cytometry analysis put forth that farnesene could decrease necrotic cell death up to 3-fold resulted from beta-amyloid exposure.
According to the investigations, farnesene can potentially be a safe, anti-necrotic and neuroprotective agents against Alzheimer's disease.
研究法呢烯倍半萜对实验性阿尔茨海默病模型的神经保护特性。
使用视黄酸将人神经母细胞瘤细胞系(SHSY-5Y)分化为神经元样细胞以构建阿尔茨海默病模型。将β-淀粉样蛋白1-42以宽剂量范围(3.125 - 200 μM)应用于转化细胞24小时和48小时以建立AD细胞毒性。然后,将法呢烯以宽光谱剂量间隔(1.625 - 100 μg/ml)应用于细胞培养物中,以研究其对β-淀粉样蛋白24小时和48小时的神经保护作用。进行3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)和乳酸脱氢酶(LDH)释放试验以确定阿尔茨海默模型中的细胞毒性。使用Hoechst 33258染色法在荧光显微镜下检查细胞的核DNA完整性。此外,分析乙酰胆碱酯酶(AChE)活性、总抗氧化能力(TAC)和总氧化状态(TOS)水平,以了解法呢烯应用于细胞培养模型的保护机制。最后,使用流式细胞术分析来找出β-淀粉样蛋白和法呢烯应用于细胞培养后细胞死亡机制。
细胞活力测试显示在24小时和48小时均对β-淀粉样蛋白毒性有显著的神经保护作用,并且Hoechst 33258荧光染色法显示在细胞培养物中应用法呢烯后坏死性死亡显著减少。最后,流式细胞术分析表明法呢烯可将β-淀粉样蛋白暴露导致的坏死性细胞死亡减少多达3倍。
根据研究,法呢烯可能是一种安全、抗坏死且对阿尔茨海默病具有神经保护作用的药物。