Telerman Alona, Ofir Rivka, Kashman Yoel, Elmann Anat
Department of Food Quality and Safety, Volcani Center, Agricultural Research Organization, POB 15159, 7528809, Rishon Lezion, Israel.
Dead Sea and Arava Science Center, Central Arava Branch, 8682500, Merkaz Sapir, Israel.
BMC Complement Altern Med. 2017 Jun 23;17(1):332. doi: 10.1186/s12906-017-1840-y.
Alzheimer's disease is a neurodegenerative disease, characterized by progressive decline in memory and cognitive functions, that results from loss of neurons in the brain. Amyloid beta (Aβ) protein and oxidative stress are major contributors to Alzheimer's disease, therefore, protecting neuronal cells against Aβ-induced toxicity and oxidative stress might form an effective approach for treatment of this disease. 3,5,4'-trihydroxy-6,7,3'-trimethoxyflavone (TTF) is a flavonoid we have purified from the plant Achillea fragrantissima; and the present study examined, for the first time, the effects of this compound on Aβ-toxicity to neuronal cells.
Various chromatographic techniques were used to isolate TTF from the plant Achillea fragrantissima, and an N2a neuroblastoma cell line was used to study its activities. The cellular levels of total and phosphorylated stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) and of total and phosphorylated extracellular signal-regulated kinase (ERK 1/2) were determined by enzyme-linked immunosorbent assay (ELISA). Intracellular reactive oxygen species (ROS) levels were measured by using 2',7'-dichlorofluorescein diacetate (DCF-DA). Cytotoxicity and cell viability were assessed by using lactate dehydrogenase (LDH) activity in cell-conditioned media, or by crystal violet cell staining, respectively.
TTF prevented the Aβ-induced death of neurons and attenuated the intracellular accumulation of ROS following treatment of these cells with Aβ. TTF also inhibited the Aβ-induced phosphorylation of the signaling proteins SAPK/JNK and ERK 1/2, which belong to the mitogen-activated protein kinase (MAPK) family.
TTF should be studied further as a potential therapeutic means for the treatment of Alzheimer's disease.
阿尔茨海默病是一种神经退行性疾病，其特征是记忆和认知功能逐渐衰退，由大脑中神经元的丧失引起。淀粉样β蛋白（Aβ）和氧化应激是阿尔茨海默病的主要促成因素，因此，保护神经元细胞免受Aβ诱导的毒性和氧化应激可能形成治疗该疾病的有效方法。3,5,4'-三羟基-6,7,3'-三甲氧基黄酮（TTF）是我们从植物香叶蓍中纯化得到的一种黄酮类化合物；本研究首次考察了该化合物对Aβ对神经元细胞毒性的影响。
采用多种色谱技术从植物香叶蓍中分离TTF，并使用N2a神经母细胞瘤细胞系研究其活性。通过酶联免疫吸附测定（ELISA）测定总应激激活蛋白激酶/c-Jun N端激酶（SAPK/JNK）和磷酸化应激激活蛋白激酶/c-Jun N端激酶以及总细胞外信号调节激酶（ERK 1/2）和磷酸化细胞外信号调节激酶的细胞水平。使用2',7'-二氯荧光素二乙酸酯（DCF-DA）测量细胞内活性氧（ROS）水平。分别通过使用细胞条件培养基中的乳酸脱氢酶（LDH）活性或结晶紫细胞染色来评估细胞毒性和细胞活力。
TTF可预防Aβ诱导的神经元死亡，并减轻Aβ处理这些细胞后ROS在细胞内的积累。TTF还抑制了Aβ诱导的属于丝裂原活化蛋白激酶（MAPK）家族的信号蛋白SAPK/JNK和ERK 1/2的磷酸化。
TTF作为治疗阿尔茨海默病的潜在治疗手段应进一步研究。
