Nazir Nausheen, Nisar Mohammad, Zahoor Muhammad, Uddin Faheem, Ullah Saeed, Ullah Riaz, Ansari Siddique Akber, Mahmood Hafiz Majid, Bari Ahmed, Alobaid Abdulrehman
Department of Biochemistry, University of Malakand, Khyber Pakhtunkhwa 18800, Pakistan.
Department of Botany, University of Malakand, Khyber Pakhtunkhwa 18800, Pakistan.
Brain Sci. 2021 Feb 19;11(2):259. doi: 10.3390/brainsci11020259.
(D. Don) is one of the endemic medicinal plants that is traditionally used to treat a number of diseases. Although the plant has been used to enhance memory, the investigational evidence supporting the nootropic effect was unsubstantial. Hence, the rationale for this study was to assess the potential beneficial effect of seed extracts on learning and memory in mice.
The powdered plant samples (aerial parts) were subjected to extraction ad fractionation. Among the extracts, crude and ethyl acetate extracts were screened for major phytochemicals through HPLC analysis. All the extracts were evaluated for the in vitro anticholinesterase (AChE and BChE) and antioxidant potentials. Among the extracts the active fraction was further assessed for improving learning and memory in mice using behavioural tests like Y-maze and novel object recognition test (NORT) using standard protocols. After behavioural tests, all the animals were sacrificed and brains tissues were assessed for the ex vivo anticholinesterase and antioxidant potentials.
Phytochemicals like chlorogenic acid, quercetin, mandelic acid, phloroglucinol, hydroxy benzoic acid, malic acid, epigallocatechin gallate, ellagic acid, rutin, and pyrogallol were identified in crude methanolic extract (Fa.Met) and ethyl acetate fraction (Fa.EtAc) through HPLC. Fa.EtAc and Fa.Chf extracts more potently inhibited AChE and BChE with IC50 values of 40 and 43 µg/mL, and 41 and 42 µg/mL, respectively. Similarly highest free radical scavenging potential was exhibited by Fa.EtAc fraction against DPPH (IC50 = 100 µg/mL) and ABTS (IC50 = 120 µg/mL). The extract doses, 100 and 200 mg/kg body weight significantly ( < 0.01) improved the short-term memory by increasing the percent spontaneous alternation in the Y-maze test along with increasing discrimination index in the NORT that clearly indicated the enhancement in the recognition memory of mice.
The extracts more potently scavenged the tested free radicals, exhibited anticholinesterase activities, improved the learning abilities and reduced the memory impairment induced by scopolamine in mice model thus suggesting that these extracts could be effectively used for the management of oxidative stress, neurodegenerative diseases and memory loss.
（D. Don）是一种传统上用于治疗多种疾病的地方性药用植物。尽管该植物已被用于增强记忆力，但支持其益智作用的研究证据并不充分。因此，本研究的目的是评估种子提取物对小鼠学习和记忆的潜在有益作用。
将植物粉末样品（地上部分）进行提取和分离。在提取物中，通过高效液相色谱（HPLC）分析对粗提物和乙酸乙酯提取物进行主要植物化学成分筛选。对所有提取物进行体外抗胆碱酯酶（乙酰胆碱酯酶和丁酰胆碱酯酶）和抗氧化潜力评估。在提取物中，使用Y迷宫和新物体识别测试（NORT）等行为测试，按照标准方案进一步评估活性成分对小鼠学习和记忆的改善作用。行为测试后，处死所有动物，评估脑组织的离体抗胆碱酯酶和抗氧化潜力。
通过HPLC在粗甲醇提取物（Fa.Met）和乙酸乙酯馏分（Fa.EtAc）中鉴定出绿原酸、槲皮素、扁桃酸、间苯三酚、羟基苯甲酸、苹果酸、表没食子儿茶素没食子酸酯、鞣花酸、芦丁和连苯三酚等植物化学成分。Fa.EtAc和Fa.Chf提取物对乙酰胆碱酯酶和丁酰胆碱酯酶的抑制作用更强，IC50值分别为40和43μg/mL，以及41和42μg/mL。同样，Fa.EtAc馏分对DPPH（IC50 = 100μg/mL）和ABTS（IC50 = 120μg/mL）表现出最高的自由基清除潜力。提取物剂量为100和200mg/kg体重时，通过增加Y迷宫测试中的自发交替百分比以及提高NORT中的辨别指数，显著（<0.01）改善了短期记忆，这清楚地表明小鼠的识别记忆得到了增强。
提取物更有效地清除了测试自由基，表现出抗胆碱酯酶活性，提高了学习能力，并减少了东莨菪碱诱导的小鼠记忆损伤，因此表明这些提取物可有效地用于管理氧化应激、神经退行性疾病和记忆丧失。
