目的 探索黑果腺肋花楸抗腦缺血的主要活性成分及其作用機制。方法 使用SwissTargetPrediction和GeneCards等數(shù)據(jù)庫篩選黑果腺肋花楸入血成分關聯(lián)靶點與腦缺血相關靶點。使用Metascape對交集靶點進行基因本體（GO）和京都基因與基因組百科全書（KEGG）通路富集分析，通過STRING數(shù)據(jù)庫與Cytoscape軟件構建并分析蛋白質-蛋白質相互作用（PPI）網(wǎng)絡?；诜肿訉訉χ匾煞趾桶悬c間相互作用進行驗證。進一步通過大鼠永久性中動脈阻塞（pMCAO）的在體模型和氧糖剝奪（OGD）處理的體外培養(yǎng)的小膠質細胞模型驗證黑果腺肋花楸活性成分花青素的抗腦缺血作用。結果 網(wǎng)絡藥理學分析結果表明花青素可能是黑果腺肋花楸發(fā)揮抗腦缺血作用的主要活性成分；GO及KEGG富集分析顯示，腫瘤壞死因子（TNF）信號通路可能為黑果腺肋花楸抗腦缺血作用的主要機制；分子對接結果顯示，基質金屬蛋白酶2（MMP-2）與花青素的結合活性較高。整體實驗結果表明，連續(xù)7 d ig給予花青素（100、200 mg·kg^-1）能夠顯著緩解pMCAO模型大鼠缺血24 h后的血腦屏障損傷，并顯著降低MMP-2表達、顯著增加VE-cadherin和occludin的表達，顯著降低炎癥因子白細胞介素-1β（IL-1β）、TNF-α、IL-6及誘導型一氧化氮合酶（iNOS）的表達。體外實驗結果表明，花青素（20、60 μg·mL^-1）能夠顯著降低OGD后BV2小膠質細胞的亞硝酸鹽含量及炎癥因子TNF-α、IL-1β、IL-6及iNOs的表達。結論 花青素可能是黑果腺肋花楸提取物抗腦缺血的主要活性成分?；ㄇ嗨啬軌蝻@著緩解大鼠永久性腦缺血后的血腦屏障損傷并抑制炎癥因子的釋放。

Objective To explore the specific components involved and underlying mechanisms of Aronia melanocarpa against cerebral ischemia. Methods SwissTargetPrediction and GeneCards databases was used to screen the blood-entry components and brain ischemia-related targets of A. melanocarpa. Gene ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was performed on the intersection targets using Metascape. The protein-protein interaction (PPI) network was constructed and analyzed using the STRING database and Cytoscape software. The interaction between the important components and targets were determined using molecular docking. Finally, the anti-cerebral ischemia effects of A. melanocarpa active component anthocyanin were evaluated using an in vivo model of rat permanent middle cerebral artery occlusion (pMCAO) and an in vitro model of cultured microglia subjected to oxygen-glucose deprivation (OGD). Results The network pharmacological analysis demonstrated that anthocyanin may serve as an effective component of A. melanocarpa in the treatment of cerebral ischemia. GO and KEGG enrichment analysis revealed that the TNF signaling pathway might be the main mechanism underlying the anti-cerebral ischemic effect of A. melanocarpa. Molecular docking results indicated a strong binding activity between anthocyanin and matrix metalloproteinases 2 (MMP-2). The in vivo studies demonstrated that oral administration of anthocyanin (100, 200 mg·kg^-1) for seven consecutive days significantly ameliorated blood-brain barrier damage, downregulated the expression of MMP-2 and upregulated the levels of VE-cadherin and occluding as well as downregulated the expression of inflammatory cytokines interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and inducible nitric oxide synthase (iNOS) at 24 h after cerebral ischemia in pMCAO-operated rats. In vitro experiments revealed that anthocyanins (20, 60 μg·mL^-1) effectively reduced the nitrite content and attenuated the expression of inflammatory factors TNF-α, IL-1β, IL-6, and iNOs in OGDtreated BV2 microglia. Conclusion The extract of A. melanocarpa may contain anthocyanin as the active component against cerebral ischemia. Anthocyanin exhibits significant potential in mitigating blood-brain barrier damage and suppressing the release of inflammatory factors following permanent cerebral ischemia in rats. The present study provides an experimental foundation for further elucidating the active components, targets, and associated mechanisms underlying the anti-cerebral ischemic effects of A. melanocarpa and anthocyanin.

R285.5

國家自然科學基金青年基金項目（82204685）;遼寧省自然科學基金面上項目（2022-MS-246）;遼寧中醫(yī)藥大學大學生創(chuàng)新創(chuàng)業(yè)訓練計劃項目（202310162004x）