目的 利用網(wǎng)絡(luò)藥理學(xué)、分子對接技術(shù)和細(xì)胞實(shí)驗(yàn)驗(yàn)證，探討藏紅花素改善阿爾茨海默?。ˋlzheimer’s disease，AD）的作用機(jī)制。方法 運(yùn)用Super-PRED、DisGenet、Genecards、Uniprot、STRING數(shù)據(jù)庫構(gòu)建藏紅花素與AD的相互作用網(wǎng)絡(luò)，導(dǎo)入Cytoscape軟件根據(jù)連接程度篩選樞紐基因，并進(jìn)行基因本體（gene ontology，GO）功能及京都基因與基因組百科全書（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析，應(yīng)用Autodock vina軟件進(jìn)行分子對接驗(yàn)證。體外采用脂多糖（lipopolysaccharide，LPS）誘導(dǎo)BV2小膠質(zhì)細(xì)胞炎癥模型，給予藏紅花素干預(yù)后，采用CCK-8法檢測細(xì)胞活力，qRT-PCR檢測腫瘤壞死因子-α（tumor necrosis factor-α，TNF-α）、白細(xì)胞介素-1β（interleukin-1β，IL-1β）、IL-6 mRNA表達(dá)，Western blotting檢測磷脂酰肌醇3-激酶（phosphatidylinositol 3-kinase，PI3K）/蛋白激酶B（protein kinase B，Akt）/糖原合成酶激酶-3β（glycogen synthase kinase-3β，GSK-3β）通路相關(guān)蛋白表達(dá)。結(jié)果 網(wǎng)絡(luò)藥理學(xué)結(jié)果顯示，藏紅花素可能通過調(diào)控PI3K-Akt信號通路、缺氧誘導(dǎo)因子-1（hypoxia inducible factor-1，HIF-1）信號通路等，作用于熱休克蛋白90AA1（heat shock protein 90 alpha family class A member 1，HSP90AA1）、核因子-κB（nuclear factor-κB，NF-κB）、Toll樣受體4（Toll-like receptor 4，TLR4）、磷脂酰肌醇3-激酶調(diào)節(jié)亞基1（phosphatidylinositol 3-kinase regulatory subunit 1，PIK3R1）、磷脂酰肌醇3-激酶催化亞基α（phosphatidylinositol 3-kinase catalytic subunit α，PIK3CA）、E1A結(jié)合蛋白p300（E1A-binding protein p300，EP300）等靶點(diǎn)，發(fā)揮治療AD的作用。分子對接結(jié)果顯示藏紅花素與PIK3R1靶點(diǎn)對接效果良好。細(xì)胞實(shí)驗(yàn)結(jié)果顯示，藏紅花素顯著抑制LPS誘導(dǎo)的BV2細(xì)胞中炎癥因子表達(dá)（P＜0.05、0.01），促進(jìn)PI3K、Akt、GSK-3β的磷酸化（P＜0.05）。結(jié)論 藏紅花素具有改善小膠質(zhì)細(xì)胞極化、抗神經(jīng)炎癥的作用，其作用機(jī)制與激活PI3K/Akt/GSK-3β通路有關(guān)。

Objective To investigate the mechanism of crocin in improving Alzheimer’s disease (AD) by network pharmacology, molecular docking and cell experiments. Methods The interaction network between crocin and AD was constructed using Super PRED, DisGenet, Genecards, Uniprot and STRING databases. Hub genes were screened based on connectivity using Cytoscape software, and gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed. Molecular docking validation was performed using Autodock vina software. In vitro, a BV2 microglial inflammation model was induced by lipopolysaccharide (LPS). After intervention with crocin, cell viability was detected by CCK-8 assay, mRNA expressions of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and IL-6 was detected by qRT-PCR, Western blotting was used to detect protein expressions related to the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β) pathway. Results The results of network pharmacology showed that crocin may act on heat shock protein 90AA1 (HSP90AA1), nuclear factor-κB (NF-κB), Toll-like receptor 4 (TLR4), phosphatidylinositol 3-kinase regulatory subunit 1 (PIK3R1), and phosphatidylinositol 3-kinase catalytic subunit α (PIK3CA) and E1A binding protein p300 (EP300), regulate PI3K-Akt signaling pathway, hypoxia inducible factor-1 (HIF-1) signaling pathway, thereby exert therapeutic effects on AD. The molecular docking results showed that crocin had a good docking effect with PIK3R1. The cell experiment results showed that crocin significantly inhibited the expressions of inflammatory factors in LPS-induced BV2 cells (P < 0.05, 0.01), and promoted the phosphorylation of PI3K, Akt and GSK-3β (P < 0.05). Conclusion Crocin has the effects of improving microglial polarization and anti-neuroinflammation, and its mechanism is related to the activation of PI3K/Akt/GSK-3β pathway.

R285.5

江蘇省自然科學(xué)基金青年項(xiàng)目（BK20240740）；南京中醫(yī)藥大學(xué)養(yǎng)老服務(wù)與管理學(xué)院專項(xiàng)研究項(xiàng)目（2024YLFWYGL012）