背景:苦参(L.)Gaertn由于其抗炎作用,在许多亚洲国家被用作民间药物,抗氧化剂,以及其他一些促进健康的特性。它还用于改善神经认知功能和缓解炎症。
目的:氧化应激和神经炎症在神经退行性疾病中起着至关重要的作用。因此,根据民族医学的说法,本研究调查了苦参种子提取物(ft-ext)对ACR诱导的神经毒性的神经保护功效。
方法:使用高效液相色谱法进行ft-ext的植物化学表征。进行来自ft-ext的鉴定化合物的分子对接。进行体外蛋白质变性测定以检查抗炎活性。将受精后5天的斑马鱼幼虫暴露于有或没有ft-ext的1mM和2.5mMACR中72小时,以研究其神经保护功效。进行实时聚合酶链反应和蛋白质印迹研究,分别分析氧化应激相关基因和蛋白质表达。
结果:提取物显示存在绿原酸,芦丁,咖啡酸,vitexin,丁香酸,槲皮素,对香豆酸,山奈酚,和阿魏酸.ft-ext的体外蛋白质变性测定显示出有效的抗炎作用。ft-ext改善了ACR介导的运动缺陷,并降低了幼虫的总死亡率。脑脂质过氧化和蛋白质羰基化结果显示ACR治疗组的氧化应激水平升高,在ft-ext处理的幼虫中减少了。提取物处理增加了nrf2,gpx的表达,和hmox1a,同时下调暴露于ACR的幼虫大脑中的trxr2水平。治疗还显示Gsk-3β失活,从而维持Nrf2和β-catenin的正常池。鉴定的ft-ext化合物的分子对接显示可能的氢和与Gsk-3β的疏水相互作用。
结论:ft-ext通过抑制Gsk-3β介导的氧化应激来预防ACR介导的神经毒性。
BACKGROUND: Fagopyrum tataricum (L.) Gaertn is used as a folk medicine in many Asian countries due to its anti-inflammatory, antioxidant, and several other health-promoting properties. It is also prescribed to improve neurocognitive functions and alleviate inflammatory conditions.
OBJECTIVE: Oxidative stress and neuroinflammation plays a crucial role in neurodegenerative conditions. Hence, based on the ethnomedical claims and available literature, the present study investigated neuroprotective efficacy of a seed extract (ft-ext) of Fagopyrum tataricum against acrylamide (ACR)-induced neurotoxicity.
METHODS: The phytochemical characterization of ft-ext was performed by a high-performance liquid chromatography method. Molecular interactions of the identified compounds of ft-ext were studied using an in-silico docking tool. An in-vitro protein denaturation assay was done to check anti-inflammatory activity. The 5 days\' post-fertilized zebrafish larvae were exposed to 1 mM and 2.5 mM ACR with or without ft-ext for 72 h to study its neuroprotective efficacy. Real-time polymerase chain reaction and western blotting studies were performed to analyse the oxidative stress-related gene and protein expressions respectively.
RESULTS: The extract showed the presence of chlorogenic acid, rutin, caffeic acid, vitexin, syringic acid, quercetin, p-coumaric acid, kaempferol, and ferulic acid. In-vitro protein denaturation assay of ft-ext showed a potent anti-inflammatory effect. The ft-ext improved ACR-mediated locomotor deficit and reduced overall mortality in the larvae. The brain lipid peroxidation and protein carbonylation results revealed an elevated level of oxidative stress in the ACR-treated group, which was reduced in ft-ext-treated larvae. The extract treatment increased the expression of nrf2, gpx, and hmox1a, while simultaneously downregulated trxr2 levels in the brain of larvae exposed to ACR. The treatment also showed inactivation of Gsk-3β, thus maintaining a normal pool of Nrf2 and β-catenin. Molecular docking of identified compounds of ft-ext showed possible hydrogen and hydrophobic interactions with Gsk-3β.
CONCLUSIONS: The ft-ext prevents ACR-mediated neurotoxicity by suppressing Gsk-3β mediated oxidative stress.