血管性认知障碍（Vascular cognitive impairment，VCI）是由脑血管疾病及其危险因素引发的临床综合征，其核心特征为至少一个认知领域的功能性损伤。其涵盖的认知障碍范围广泛，从轻度认知损伤到痴呆。VCI的主要症状包括注意力缺陷、记忆力减退、语言障碍、执行功能障碍和空间认知能力下降，这些症状严重影响患者的生活质量。随着人口老龄化的增长，VCI的患病率迅速上升，成为仅次于阿尔茨海默病的第二大认知缺陷类型。

目前，临床主要通过调控血管危险因素和脑血管改善认知功能，常用药物包括胆碱酯酶抑制剂（如多奈哌齐）、钙离子通道拮抗剂（如尼莫地平）及血管紧张素转换酶抑制剂（如卡托普利）等。然而，现有治疗药物的临床疗效仍存在争议。因此、探寻安全、有效、低毒的治疗药物迫在眉睫。

传统中药作为创新药物发现的重要源泉，具有多成分、多靶点、安全性高等特点，在慢性病防治方面具有独特优势，为VCI的治疗策略提供新方向。五味子作为传统益智安神类中药，已有数千年药用历史。现代药理研究证实，其活性成分具有神经保护、抗炎、抗氧化、保肝降酶等多种功效。中药五味子包含两种基原植物，分别为五味子（Schisandra chinensis，SCF）和华中五味子（Schisandra sphenanthera，SSF），二者传统功效一致。然而，自2000版《中国药典》开始，二者作为独立药材收录，分别将SCF和SSF记录为北五味子和南五味子，并建立了差异化的质量控制体系。

虽然SCF和SSF在神经保护方面具有相同的传统功效记载，然而在改善认知障碍的药效差异及潜在未见报道。本研究针对SCF和SSF，通过体内外实验系统比较它们对VCI的改善作用，并确定其活性部位；进一步通过代谢组学与转录组学联合分析，探讨其分子调控机制；基于课题组前期对活性部位的化学成分研究，结合网络药理学和体外活性评价，确定其活性成分。具体研究内容与结果如下：

1. SCF和SSF总提物改善VCI药效比较

基于脂多糖（Lipopolysaccharide，LPS）刺激及氧糖剥夺/复糖复氧（Oxygen-glucose deprivation/reoxygenation，OGD/R）诱导的BV2细胞模型，开展体外活性评价。实验结果显示，40 μg/mL剂量下SCF较SSF显著抑制LPS诱导的炎症因子（NO、TNF-α、IL-6）水平（p < 0.01）。同时，SCF较SSF显著改善OGD/R处理后的细胞活力及线粒体膜电位（Mitochondrial membrane potential，MMP）损伤（p < 0.01）。进一步采用双侧颈总动脉狭窄（Bilateral common carotid artery stenosis，BCAS）诱导的VCI小鼠模型进行体内药效评价，实验结果与体外活性评价一致，400 mg/kg剂量下，SCF较SSF可显著改善BCAS小鼠学习记忆能力，恢复脑血流量，改善白质脱髓鞘病变（p < 0.05）。 

2. SCF改善VCI活性部位的确定

采用不同极性的溶剂对SCF醇提物依次萃取，分别获得石油醚部位（Petroleum ether，PEF）、乙酸乙酯部位（Ethyl acetate，ETA）及大极性部位（H₂O soluble fractions，HOF）。体外活性评价发现，PEF（40 μg/mL）可显著抑制LPS诱导的BV2细胞炎症因子的生成（p < 0.01），并改善OGD/R处理后的细胞活力及MMP损伤（p < 0.01），而ETA和HOF无显著效果。进一步采用BCAS小鼠模型，验证PEF活性部位对认知功能、脑血流及白质损伤的改善作用。实验结果表明，PEF可剂量依赖性改善BCAS小鼠认知功能障碍，恢复脑血流量，减少白质区域髓鞘损伤（p < 0.05）。

3. PEF改善VCI作用机制初探

采用全脑代谢组学结合海马转录组学进一步探讨PEF改善VCI的作用机制。代谢组学结果显示，花生四烯酸代谢及谷胱甘肽代谢通路在海马与岛叶脑区富集，提示其通过调节氧化应激和神经炎症影响记忆、认知、内脏感觉和运动等大脑功能。嘌呤代谢通路在胼胝体与杏仁核脑区富集，提示其通过能量代谢影响脑内协调沟通、情绪、学习和记忆功能。转录组学数据显示，PEF通过上调胶质细胞相关的髓鞘再生，抑制神经炎症，形成多细胞层级的协同修复机制。其神经保护作用与调控多巴胺能和谷氨酸能突触功能密切相关。

4. 基于网络药理学结合体外活性评价确定PEF改善VCI的潜力活性化合物

课题组前期从PEF中共鉴定66个化合物，主要包括木脂素类化合物（50种联苯环辛烯型，4种二芳基丁烷型及3种四氢呋喃型），此外还发现4种萜类、3种生物碱及2种黄酮类化合物。基于鉴定的化学成分，采用网络药理学预测获得PEF中发挥药效的15个活性成分，其关键作用靶点包括SRC、MAPK1、AKT1等，涉及PI3K-AKT、FoxO等通路。采用LPS刺激及OGD/R处理的BV2细胞模型，进一步对15个活性化合物进行体外验证，发现Schisandrin A、Schisandrin B、Schisantherin D、Gomisin J等8个化合物显著抑制NO生成（p < 0.05），并改善MMP损伤，其中Schisantherin D（20 μM）活性优于阳性对照。

综上，SCF较SSF显著改善VCI，其活性主要集中在PEF，核心活性成分包括Schisantherin D、Gomisin J等。其作用机制可能通过作用SRC、MAPK1、AKT1等靶点，调控PI3K-AKT/FoxO通路改善神经炎症、氧化应激及能量代谢，进而改善VCI病理进程。

Vascular cognitive impairment (VCI) is a clinical syndrome caused by cerebrovascular diseases and their risk factors, characterized by functional impairment in at least one cognitive domain as its core feature. This spectrum of cognitive disorders ranges from mild cognitive impairment to dementia. The main symptoms of VCI include attention deficits, memory loss, language impairment, executive dysfunction, and decline in spatial cognitive abilities, which significantly impact patients' quality of life. With advancing age, the prevalence of VCI increases rapidly, making it the second most common type of cognitive deficit after Alzheimer's disease.

Currently, clinical approaches to improve cognitive function focus on managing vascular risk factors and cerebrovascular diseases. Commonly used medications include cholinesterase inhibitors (e.g., Donepezil), calcium channel blockers (e.g., Nimodipine), and angiotensin-converting enzyme inhibitors (e.g., Captopril). However, the clinical efficacy of existing treatment regimens remains controversial. Notably, traditional Chinese medicine, with its multi-target mechanisms, systemic regulation, and high safety profile, exhibits unique advantages in the prevention and management of chronic diseases, offering novel directions for VCI therapeutic strategies.

Wuweizi, a well-known TCM, has been used as a tonic and sedative agent for thousands of years. Modern pharmacological studies have suggested that Wuweizi possesses a range of biological activities, such as neuroprotective, anti-inflammatory, antioxidative, and hepatoprotective properties. Historically, Wuweizi is referred to as the ripe fruit of both Schisandra chinensis (SCF) and Schisandra sphenanthera (SSF). However, since the 2000 edition of Chinese Pharmacopoeia, SCF and SSF have been classified as distinct crude drugs with independent quality standards, despite similarities in their described efficacies in ChP.

Although SCF and SSF share identical neuroprotective effects, their differential efficacy in improving cognitive impairment remains unreported. This study systematically compared the therapeutic effects of SCF and SSF on VCI through in vivo and in vitro experiments to identify their active fractions. The molecular regulatory mechanisms were explored through integrated metabolomics and transcriptomics analysis. Additionally, the chemical profiles of these active fractions were previously characterized by the research group, and their bioactive compounds were further determined by integrating network pharmacology with in vitro activity validation. The specific research content and results are as follows:

1. Comparative Evaluation of Total Extracts from SCF and SSF in Improving VCI

Based on in vitro models of BV2 microglial cells induced by lipopolysaccharide (LPS) stimulation and oxygen-glucose deprivation/reoxygenation (OGD/R), the pharmacological effects were evaluated. Results demonstrated that at 40 μg/mL, SCF significantly inhibited LPS-induced inflammatory factors (nitric oxide, tumour necrosis factor-α, and interleukin-6) compared to SSF (p < 0.01). Concurrently, SCF markedly ameliorated OGD/R-induced cell viability loss and mitochondrial membrane potential (MMP) damage relative to SSF (p < 0.01). Further in vivo efficacy evaluation using a VCI mouse model induced by bilateral common carotid artery stenosis (BCAS) corroborated the in vitro findings. At 400 mg/kg, SCF outperformed SSF in significantly improving learning and memory performance, restoring cerebral blood flow, and mitigating white matter demyelination in BCAS mice (p < 0.05).

2. Identification of Active Fraction in SCF for Improving VCI

The ethanol extract of SCF was sequentially partitioned using solvents of varying polarities to obtain three fractions: petroleum ether fraction (PEF), ethyl acetate fraction (ETA), and water-soluble fraction (HOF). In vitro activity evaluation revealed that PEF (40 μg/mL) significantly inhibited LPS-induced inflammatory factor production in BV2 microglial cells (p < 0.01) and ameliorated OGD/R-induced cell viability loss and MMP damage (p < 0.01), while ETA and HOF showed no significant effects. Further validation using the BCAS-induced mouse model confirmed the therapeutic potential of the PEF on cognitive function, cerebral blood flow, and white matter pathology. Experimental results demonstrated that PEF dose-dependently ameliorated cognitive dysfunction, restored cerebral blood flow, and reduced myelin damage in white matter regions of BCAS mice (p < 0.05).

3. Preliminary Exploration of PEF’s Mechanisms in Improving VCI

Integrated whole-brain metabolomics and hippocampal transcriptomics were employed to investigate the therapeutic mechanisms of PEF in VCI. Metabolomic analysis revealed significant enrichment of arachidonic acid metabolism and glutathione metabolic pathways in the hippocampus and insular cortex, suggesting their regulatory roles in memory, cognition, visceral sensation, and motor functions through modulating oxidative stress and neuroinflammation. Additionally, the purine metabolism pathway was predominantly enriched in the corpus callosum and amygdala, indicating its involvement in brain coordination, emotional regulation, learning and memory via energy metabolism modulation. Transcriptomic data demonstrated that PEF promotes a multi-cellular synergistic repair mechanism by upregulating glial cell-mediated myelin regeneration while suppressing neuroinflammation. The neuroprotective effects were closely associated with the regulation of dopaminergic and glutamatergic synaptic functions.

4. Identification of Potential Active Compounds in PEF for Improving VCI via Network Pharmacology and In Vitro Activity Validation

The research team previously identified a total of 66 compounds from PEF, primarily including lignans (50 dibenzocyclooctene types, 4 diarylbutane types, and 3 tetrahydrofuran types). Additionally, 4 terpenoids, 3 alkaloids, and 2 flavonoids were also discovered. Based on the identified chemical components, network pharmacology analysis predicted 15 active ingredients in PEF responsible for pharmacological effects. Key targets included SRC, MAPK1, and AKT1, involving pathways such as PI3K-AKT and FoxO. Further in vitro validation of the 15 active compounds was conducted using BV2 cell models subjected to LPS stimulation and OGD/R treatment. Eight compounds, including Schisandrin A, Schisandrin B, and Schisantherin D, significantly inhibited NO production (p < 0.05) and ameliorated MMP impairment. Notably, Schisantherin D (20 μM) exhibited superior activity compared to the positive control.

In summary, SCF demonstrates significantly superior efficacy over SSF in ameliorating VCI, with its bioactive effects primarily concentrated in the PEF. Core active compounds include Schisantherin D, Gomisin J, and related compounds. The therapeutic mechanisms likely involve targeting key nodes such as SRC, MAPK1, and AKT1 to modulate PI3K-AKT/FoxO signaling pathways, thereby alleviating neuroinflammation, oxidative stress, and energy metabolism dysregulation, ultimately mitigating VCI pathological progression.