Zhang Ting, Park Sunmin
Key Laboratory of Biological Resources and Ecology of Pamirs Plateau in Xinjiang Uygur Autonomous Region, College of Life and Geographic Sciences, Kashi University, Kashi 844000, China.
Department of Bioconvergence, Hoseo University, Asan 31499, Republic of Korea.
Int J Mol Sci. 2025 Sep 3;26(17):8545. doi: 10.3390/ijms26178545.
Neuroinflammation plays a central role in the pathogenesis of Alzheimer's disease (AD), with amyloid-β (Aβ) deposition and neurofibrillary tangles driving both central and peripheral inflammatory responses. This study investigated the neuroprotective and anti-inflammatory effects of (VT), (PM), Folium (AVF), and (EF) using network pharmacology and a co-culture model of PC12 neuronal and Caco-2 intestinal epithelial cells. Bioactive compounds were identified via high-performance liquid chromatography (HPLC) and screened with network pharmacology analysis, yielding 27 for VT, 10 for PM, 6 for AVF, and 3 for EF. Molecular docking confirmed strong binding affinities between the key bioactive compounds and AD-related targets. A co-culture system of PC12 neuronal and Caco-2 intestinal epithelial cells was established to evaluate the effects of VT, PM, AVF, and EF extracts (at concentrations of 10 µg/mL, 20 µg/mL, and 50 µg/mL) and donepezil hydrochloride (positive-control) on Aβ-induced neurotoxicity and lipopolysaccharide (LPS)-induced intestinal inflammation, to assess cell viability, and effects on oxidative stress, mitochondrial function, and inflammatory markers. The VT, PM, AVF, and EF extracts activated phosphoinositide 3-kinase (PI3K)-Akt-glycogen synthase kinase-3β (GSK-3β) signaling, enhanced phosphorylation of AMP kinase, suggesting inhibition of Aβ accumulation and tau hyperphosphorylation ( < 0.05). However, donepezil hydrochloride only enhanced AMPK phosphorylation. The extracts reduced lipid peroxidation and acetylcholinesterase by about 5-fold. JC-1 staining confirmed preserved mitochondrial membrane potential, while hematoxylin and eosin staining indicated improved intestinal barrier integrity ( < 0.05). PM and AVF reduced the number of mast cells ( < 0.05). In conclusion, these findings highlight the multi-target potential of VT, PM, AVF, and EF in mitigating both neuronal and intestinal inflammation. Their dual regulatory effects on the gut-brain axis suggest promising therapeutic applications in AD through the modulation of central and peripheral immune responses.
神经炎症在阿尔茨海默病（AD）的发病机制中起核心作用，淀粉样β蛋白（Aβ）沉积和神经纤维缠结驱动中枢和外周炎症反应。本研究使用网络药理学以及PC12神经元和Caco-2肠上皮细胞的共培养模型，研究了长春花（VT）、太子参（PM）、青蒿叶（AVF）和杜仲（EF）的神经保护和抗炎作用。通过高效液相色谱（HPLC）鉴定生物活性化合物，并进行网络药理学分析筛选，长春花得到27种，太子参得到10种，青蒿叶得到6种，杜仲得到3种。分子对接证实了关键生物活性化合物与AD相关靶点之间具有强结合亲和力。建立PC12神经元和Caco-2肠上皮细胞的共培养系统，以评估长春花、太子参、青蒿叶和杜仲提取物（浓度为10μg/mL、20μg/mL和50μg/mL）以及盐酸多奈哌齐（阳性对照）对Aβ诱导的神经毒性和脂多糖（LPS）诱导的肠道炎症的影响，评估细胞活力以及对氧化应激、线粒体功能和炎症标志物的影响。长春花、太子参、青蒿叶和杜仲提取物激活了磷酸肌醇3激酶（PI3K）-蛋白激酶B（Akt）-糖原合成酶激酶-3β（GSK-3β）信号通路，增强了AMP激酶的磷酸化，表明抑制了Aβ积累和tau蛋白过度磷酸化（P<0.05）。然而，盐酸多奈哌齐仅增强了AMPK的磷酸化。提取物使脂质过氧化和乙酰胆碱酯酶降低了约5倍。JC-1染色证实线粒体膜电位得以保留，而苏木精和伊红染色表明肠道屏障完整性得到改善（P<0.05）。太子参和青蒿叶减少了肥大细胞数量（P<0.05）。总之，这些发现突出了长春花、太子参、青蒿叶和杜仲在减轻神经元和肠道炎症方面的多靶点潜力。它们对肠-脑轴的双重调节作用表明，通过调节中枢和外周免疫反应，在AD治疗中具有广阔的应用前景。
