Institute of Chinese Materia Medica, The MOE Key Laboratory for Standardization of Chinese Medicines and The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Rood, Shanghai, 201203, China.
Institute of Chinese Materia Medica, The MOE Key Laboratory for Standardization of Chinese Medicines and The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Rood, Shanghai, 201203, China.
J Ethnopharmacol. 2024 Jan 10;318(Pt A):116792. doi: 10.1016/j.jep.2023.116792. Epub 2023 Jun 24.
Fufangmuniziqi formula (FFMN), a traditional Uyghur medicine used in China, is derived from an ancient Uyghur medical book and consists of 13 herbs. The herbs of FFMN, such as Peganum harmala L., Glycyrrhiza uralensis Fisch., and Nigella glandulifera, have been demonstrated to have acetylcholinesterase (AChE) inhibitory, anti-neuroinflammatory, or antioxidant effects. Therefore, FFMN may have a good anti-Alzheimer's disease (AD) effect, but its specific action and mechanism need to be further proven.
This study aims to investigate the anti-AD effects of FFMN and the role played by alkaloids, flavonoids, and saponins in anti-AD.
The alkaloids, flavonoids, and saponins fractions of FFMN were prepared by macroporous resin chromatography. The absorbed ingredients in the drug-containing serum were identified by UPLC⁃Q⁃TOF⁃MS. An AD mouse model was established by intraperitoneal injection of scopolamine (SCO). The role of different fractions of FFMN in the anti-AD process was examined by Morris water maze (MWM), in-vitro cell, and AChE inhibition assay.
A total of 20 ingredients were identified in the serum samples collected after oral administration of FFMN, and seven compounds were selected as candidate active compounds. MWM experiments showed that different fractions of FFMN could significantly improve SCO-induced learning memory impairment in mice. The alkaloids fraction (ALK) regulated cholinergic function by inhibiting AChE activity, activating choline acetyltransferase activity, and protein expression. Flavonoids and saponins were more potent than the ALK in downregulating pro-inflammatory factors or inflammatory mediators, such as TNF-α, MPO, and nitric oxide. Western blot results further confirmed that flavonoids and saponins attenuated neuroinflammation by inhibiting the phosphorylation of IκB and NF-κB p65. This result was also verified by in-vitro cellular assays. FFMN enhanced antioxidant defense by increasing the activity of superoxide dismutase and reducing the production of MDA. Combined with cellular experiments, flavonoids and saponins were proven more protective against oxidative damage.
FFMN improved cognitive and memory impairment in the SCO-induced AD mouse model. ALK mainly enhanced the function of the cholinergic system. Flavonoid and saponin fractions mainly attenuated neuroinflammation and oxidative stress by modulating the NF-κB pathway. All these findings strongly suggested that the combination of alkaloid, flavonoid, and saponin fractions derived from FFMN is a promising anti-AD agent that deserves further development.
复方木尼孜其颗粒（FFMN）是一种源自中国古代维吾尔医学书籍的传统维吾尔药物，由 13 种草药组成。FFMN 的草药，如骆驼蓬、甘草和黑种草，已被证明具有乙酰胆碱酯酶（AChE）抑制、抗神经炎症或抗氧化作用。因此，FFMN 可能对阿尔茨海默病（AD）有很好的治疗作用，但具体作用机制尚需进一步证实。
本研究旨在探讨 FFMN 的抗 AD 作用及其生物碱、黄酮类和皂苷在抗 AD 中的作用。
采用大孔树脂色谱法制备 FFMN 的生物碱、黄酮类和皂苷部位。采用 UPLC-Q-TOF-MS 鉴定含药血清中被吸收的成分。腹腔注射东莨菪碱（SCO）建立 AD 小鼠模型。通过 Morris 水迷宫（MWM）、体外细胞和 AChE 抑制试验考察 FFMN 各部位在抗 AD 过程中的作用。
共鉴定出 FFMN 灌胃后血清样本中的 20 种成分，选择 7 种化合物作为候选活性化合物。MWM 实验表明，FFMN 的不同部位可显著改善 SCO 诱导的小鼠学习记忆障碍。生物碱部位（ALK）通过抑制 AChE 活性、激活胆碱乙酰转移酶活性和蛋白表达来调节胆碱能功能。黄酮类和皂苷的抗炎作用强于 ALK，可下调 TNF-α、MPO 和一氧化氮等促炎因子或炎症介质。Western blot 结果进一步证实，黄酮类和皂苷通过抑制 IκB 和 NF-κB p65 的磷酸化来减轻神经炎症。这一结果也通过体外细胞试验得到了验证。FFMN 通过增加超氧化物歧化酶的活性和减少 MDA 的产生来增强抗氧化防御能力。结合细胞实验，黄酮类和皂苷对氧化损伤具有更好的保护作用。
FFMN 改善了 SCO 诱导的 AD 小鼠模型的认知和记忆障碍。ALK 主要增强胆碱能系统的功能。黄酮类和皂苷部位主要通过调节 NF-κB 通路来减轻神经炎症和氧化应激。所有这些发现都强烈表明，FFMN 中的生物碱、黄酮类和皂苷联合应用是一种很有前途的抗 AD 药物，值得进一步开发。
