Abstract

Objective

To investigate neuroprotective properties of the farnesene sesquiterpene on the experimental Alzheimer’s disease model in vitro.

Methods

Human neuroblastoma cell line (SHSY-5Y) was differentiated into neuron-like cells by using retinoic acid to constitute the in vitro 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.

Results

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.

Conclusion

According to the investigations, farnesene can potentially be a safe, anti-necrotic and neuroprotective agents against Alzheimer’s disease.

中文翻译：

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法呢烯倍半萜对阿尔茨海默病分化神经母细胞瘤模型的体外神经保护作用

摘要

客观的

研究法呢烯倍半萜对体外实验性阿尔茨海默病模型的神经保护特性。

方法

体外利用维甲酸构建人神经母细胞瘤细胞株（SHSY-5Y）分化成神经元样细胞阿尔茨海默病模型。将 β-淀粉样蛋白 1-42 蛋白以宽剂量范围 (3.125-200 μM) 应用于转化细胞 24 和 48 小时以建立 AD 细胞毒性。然后，将法呢烯以广谱剂量间隔（1.625-100 μg/ml）应用于细胞培养物，以研究 24 和 48 小时对 β-淀粉样蛋白的神经保护作用。执行 3-(4,5-二甲基-噻唑-2-基) 2,5-二苯基溴化四唑 (MTT) 和乳酸脱氢酶 (LDH) 释放测试以确定阿尔茨海默氏症模型中的细胞毒性。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，乙酰胆碱酯酶 (AChE) 活性，分析总抗氧化能力（TAC）和总氧化状态（TOS）水平，以了解法呢烯在细胞培养模型中的应用保护机制。最后，流式细胞仪分析用于找出β-淀粉样蛋白和法呢烯应用于细胞培养物后的细胞死亡机制。

结果

细胞活力测试显示在 24 和 48 小时内对 β-淀粉样蛋白毒性有显着的神经保护作用，并且 Hoechst 33258 荧光染色方法显示在细胞培养物中施用法呢烯后坏死性死亡显着减少。最后，流式细胞术分析表明，法呢烯可将因暴露于β-淀粉样蛋白而导致的坏死细胞死亡减少 3 倍。

结论

根据调查，法呢烯可能是一种安全、抗坏死和神经保护剂，可对抗阿尔茨海默病。