目的 运用脂质代谢组学和网络药理学寻找川陈皮素对胃癌顺铂耐药细胞（AGS-DDP）的核心靶点，并结合细胞实验进一步探究川陈皮素逆转胃癌顺铂耐药的机制。方法 采用UPLC-Orbitrap质谱系统进行非靶向脂质组学分析。利用GENE Card、TCMSP数据库挖掘川陈皮素和甘油磷脂代谢通路的潜在靶点，通过微生信在线作图工具平台整合二者的交集靶点，并进行基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析，通过STRING数据库绘制蛋白质-蛋白质相互作用（protein-protein interaction，PPI）网络，运用Cytoscape软件将川陈皮素的核心靶点可视化。川陈皮素给药后，通过CCK-8法检测AGS和AGS-DDP细胞的存活率；流式细胞术检测AGS-DDP细胞的凋亡情况；划痕实验检测AGS-DDP细胞的迁移能力；Western blotting检测AGS-DDP细胞的B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2）、Bcl-2相关X蛋白（Bcl-2 associated X protein，Bax）、E-cadherin、N-cadherin、Vimentin、过氧化物酶体增殖物激活受体γ（peroxisome proliferators-activated receptor γ，PPARγ）和三磷酸腺苷柠檬酸裂解酶（adenosine triphosphate citrate lyase，ACLY）蛋白表达。结果 AGS和AGS-DDP细胞通过脂质组学分析检测得到31个亚类中的1450个差异的脂质化合物，主要富集在甘油磷脂代谢通路，对川陈皮素潜在靶点进行KEGG富集分析结果显示川陈皮素具有逆转铂类耐药的潜力。细胞实验结果显示，与对照组比较，川陈皮素剂量相关性地抑制AGS-DDP细胞的存活率（P<0.05、0.01），增加AGS-DDP细胞对顺铂的敏感性（P<0.05），且能诱导AGS-DDP细胞凋亡并抑制其迁移能力（P<0.05、0.01）。Western blotting实验结果显示川陈皮素组Bax、E-cadherin蛋白表达水平显著升高（P<0.05、0.01），Bcl-2、N-cadherin、Vimentin、ACLY和PPARγ蛋白表达水平均显著降低（P<0.05、0.01）。结论 AGS和AGS-DDP细胞的差异脂质代谢产物主要富集在甘油磷脂代谢通路，川陈皮素通过调控脂质合成氧化关键基因抑制AGS-DDP细胞增殖、迁移，最终逆转胃癌顺铂耐药。

Objective To search for the core targets of nobiletin on gastric cancer cisplatin resistant cells (AGS-DDP) by lipid metabolomics and network pharmacology, and further explore the mechanism of nobiletin on reversing gastric cancer cisplatin resistance through cell experiments. Methods UPLC-Orbitrap mass spectrometry system was used for non-targeted lipidomics analysis. GENE Card and TCMSP databases were utilized to explore potential targets for the metabolic pathways of nobiletin and glycerol phospholipids, the intersection targets were integrated through Microbioinformatics online mapping tool platform, and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis was performed. STRING database was used to construct protein-protein interaction (PPI) network, the core targets of nobiletin was visualized by Cytoscape software. After administration of nobiletin, the survival rate of AGS and AGS-DDP cells was detected by CCK-8 method; Flow cytometry was used to detect the apoptosis of AGS-DDP cells; Scratch experiment was used to detect the migration ability of AGS-DDP cells; Western blotting was used to detect B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), E-cadherin, N-cadherin, Vimentin, peroxisome proliferators-activated receptor γ (PPARγ) and adenosine triphosphate citrate lyase (ACL) protein expressions in AGS-DDP cells. Results A total of 1450 different lipid compounds were detected in 31 subclasses of AGS and AGS-DDP cells through lipidomics analysis, mainly enriched in glycerol phospholipid metabolism pathway. KEGG enrichment analysis of potential targets of nobiletin showed that nobiletin had the potential to reverse platinum resistance. The results of cell experiment showed that compared with control group, nobiletin significantly inhibited the survival rate of AGS-DDP cells in a dose-dependent manner (P < 0.05, 0.01), increased the sensitivity of AGS-DDP cells to cisplatin (P < 0.05), induced AGS-DDP cells apoptosis and inhibited its migration ability (P < 0.05, 0.01). The results of Western blotting experiments showed that Bax and E-cadherin protein expression levels were significantly increased (P < 0.05, 0.01), as well as Bcl-2, N-cadherin, Vimentin, ACL and PPARγ protein expression levels were significantly reduced (P < 0.05, 0.01). Conclusion The differential lipid metabolites between AGS and AGS-DDP cells are mainly enriched in glycerol phospholipid metabolism pathway. Nobiletin inhibits the proliferation and migration of AGS-DDP cells by regulating key genes for lipid synthesis and oxidation, ultimately reversing cisplatin resistance in gastric cancer.

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国家自然科学基金资助项目（81973609）；国家自然科学基金资助项目（81973782）；国家自然科学基金资助项目（82174197）；江苏省自然科学基金项目（BK20211392）；江苏省医学青年人才项目（QNRC2016641）；江苏省“333”计划项目（LGY2018065）；江苏省中医院学术人才计划项目（Y2021RC29，Y2021RC45）