Department of Traditional Pharmacy, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran.
Metab Brain Dis. 2019 Feb;34(1):165-172. doi: 10.1007/s11011-018-0329-9. Epub 2018 Nov 6.
Alzheimer's disease is a type of cerebrovascular problem with progressive mental disabilities for the patient. This study aimed to investigate the protective effect of safranal on toxicity and oxidative damage induced by beta-amyloid (Aβ) and hydrogen peroxide (HO) in PC12 cells as an appropriate model of Alzheimer's cell damage. PC12 cells pretreated with saffron extract (2.5-40 μg/ml), essential oil (2.5-40 μg/ml), safranal (2.5-5-40 μM) and donepezil (5, 10 and 20 μM) for 120 min. Then exposed to either Aβ (25 μM) for 48 h or HO (150 μM) for 24 h. In the end, the cell survival and intracellular reactive oxygen species (ROS) production analyzed. The anti-apoptotic effects of safranal in PC12 cells were studied using flow cytometry after PI staining. Also, western blot analysis of Cyt c, survivin, p44/42 MAPK (ERK1/2), Phospho-p44/42 MAPK (ERK1/2), PI3 Kinase P85, Phospho-PI3 Kinase P85, phospho SAPK/JNK, SAPK/JNK and caspase 3 performed for detection of apoptosis. Safranal (2.5 and 5 μM) and donepezil (10 and 20 μM) significantly decreased the Aβ toxicity. The ROS significantly attenuated when cells pretreated with essential oil, saffron extract, safranal, and donepezil. Cell apoptosis significantly increased after treatment with Aβ (25 μM) compared to control. However, after pretreatment with safranal (2.5 μM) apoptosis was significantly reduced. Western blot analysis of PC12 cells showed that 25 μM Aβ could increase proteins involved in apoptosis signaling and pretreatment with safranal (2.5 μM) could decrease the apoptosis. According to the results, safranal showed anti-apoptotic and antioxidant effects and may exert promising potential for the prevention of Alzheimer's disease.
阿尔茨海默病是一种伴有患者进行性精神障碍的脑血管问题。本研究旨在探讨藏红花素（safranal）对β-淀粉样蛋白（Aβ）和过氧化氢（HO）诱导的 PC12 细胞毒性和氧化损伤的保护作用，PC12 细胞作为阿尔茨海默病细胞损伤的合适模型。PC12 细胞用藏红花提取物（2.5-40μg/ml）、精油（2.5-40μg/ml）、藏红花素（2.5-5-40μM）和多奈哌齐（5、10 和 20μM）预处理 120min。然后暴露于 Aβ（25μM）48h 或 HO（150μM）24h。最后，分析细胞存活率和细胞内活性氧（ROS）的产生。用碘化丙啶（PI）染色后通过流式细胞术研究藏红花素在 PC12 细胞中的抗凋亡作用。还进行了 Cyt c、survivin、p44/42 MAPK（ERK1/2）、Phospho-p44/42 MAPK（ERK1/2）、PI3 Kinase P85、Phospho-PI3 Kinase P85、phospho SAPK/JNK、SAPK/JNK 和 caspase 3 的 Western blot 分析，以检测凋亡。藏红花素（2.5 和 5μM）和多奈哌齐（10 和 20μM）显著降低了 Aβ的毒性。当用精油、藏红花提取物、藏红花素和多奈哌齐预处理细胞时，ROS 显著减弱。与对照组相比，用 Aβ（25μM）处理后细胞凋亡显著增加。然而，用 2.5μM 藏红花素预处理后，凋亡明显减少。PC12 细胞的 Western blot 分析表明，25μM Aβ可以增加参与细胞凋亡信号的蛋白质，而用 2.5μM 藏红花素预处理可以减少凋亡。根据这些结果，藏红花素显示出抗凋亡和抗氧化作用，可能对预防阿尔茨海默病具有潜在的应用价值。
