目的 基于組分結(jié)構(gòu)理論及網(wǎng)絡(luò)藥理學(xué)探究綠茶有效組分（奎尼酸、表兒茶素、沒食子兒茶素、表沒食子兒茶素、沒食子兒茶素沒食子酸酯和表沒食子兒茶素沒食子酸酯）干預(yù)年齡相關(guān)性黃斑變性（AMD）的最佳配伍結(jié)構(gòu)及作用機(jī)制。方法 采用氯化鈷致斑馬魚AMD模型，結(jié)合均勻設(shè)計(jì)法，以斑馬魚眼底血管異常生長的相對(duì)抑制率為評(píng)價(jià)指標(biāo)，通過多元線性回歸分析篩選綠茶有效組分干預(yù)AMD的最佳配伍結(jié)構(gòu)；借助網(wǎng)絡(luò)藥理學(xué)預(yù)測(cè)其作用通路及關(guān)鍵靶點(diǎn)，并利用分子對(duì)接技術(shù)與實(shí)時(shí)熒光定量（qRT-PCR）法驗(yàn)證綠茶有效組分對(duì)關(guān)鍵靶點(diǎn)的調(diào)控作用。結(jié)果 當(dāng)奎尼酸、表兒茶素、沒食子兒茶素、表沒食子兒茶素、沒食子兒茶素沒食子酸酯、表沒食子兒茶素沒食子酸酯的質(zhì)量濃度分別為40、40、25、50、15、15 μg·mL^-1時(shí)干預(yù)AMD的藥效最佳，其最佳配伍結(jié)構(gòu)為8∶ 8∶ 5∶ 10∶ 3∶ 3；綠茶有效組分可能通過調(diào)控PI3K、Akt、TP53、HIF1A、VEGF等基因的表達(dá)，激活PI3K-Akt、HIF-1、Apoptosis和TNF信號(hào)通路，從而發(fā)揮干預(yù)AMD的藥效作用。結(jié)論 綠茶有效組分可協(xié)同增效發(fā)揮干預(yù)AMD作用，且存在明確的最佳配伍結(jié)構(gòu)，本研究為中藥組分配伍規(guī)律研究提供思路，為綠茶干預(yù)AMD的組分新藥研發(fā)奠定基礎(chǔ)。

Objective To explore the optimal combination structure and mechanism of action of effective components of green tea (quinic acid, epicatechin, gallocatechin, epigallocatechin, gallocatechin gallate, and epigallocatechin gallate) in the intervention of agerelated macular degeneration (AMD) based on the theory of component structure and network pharmacology. Methods A zebrafish AMD model induced by cobalt chloride was used, combined with uniform design method. The relative inhibition rate of abnormal growth of zebrafish retinal vessels was taken as the dependent variable. The optimal combination structure of green tea effective components in the intervention of AMD was screened by multiple linear regression analysis. Network pharmacology was used to predict the action pathways and key targets, and molecular docking technology and real-time fluorescence quantitative (qRT-PCR) method were used to verify the regulatory effect of green tea effective components on key targets. Results When the mass concentrations of quinic acid, epicatechin, gallocatechin, epigallocatechin, gallocatechin gallate, and epigallocatechin gallate were 40, 40, 25, 50, 15, and 15 μg·mL^-1, respectively, the intervention effect on AMD was the best, and the optimal combination structure was 8∶ 8∶ 5∶ 10∶ 3∶ 3. Green tea effective components may exert the intervention effect on AMD by regulating the expression of genes such as PI3K, Akt, TP53, HIF1A, and VEGF, and activating the PI3K-Akt, HIF-1, Apoptosis, and TNF signaling pathways. Conclusion The effective components of green tea can exert a synergistic effect in the intervention of AMD, and there is a clear optimal combination structure. This study provides ideas for the research on the combination rules of traditional Chinese medicine components and lays a foundation for the development of new component drugs of green tea for the intervention of AMD.

R285.5

山東省重點(diǎn)研發(fā)計(jì)劃項(xiàng)目（2021CXGC010511）；山東省科技型中小企業(yè)創(chuàng)新能力提升工程項(xiàng)目（2023TSGC0271）