Çelik Cansel, Özhan Yağmur, Öztürk Ceren, Dede Zulal Sevgi, Citoglu Tugce, Gungor Burcin, Aru Başak, Gurdal Enise Ece, Sippl Wolfgang, Sipahi Hande, Tekşen Mehtap, Kırmızıbekmez Hasan
Department of Pharmacognosy, Faculty of Pharmacy, Yeditepe University, TR-34755, Kayışdağı, İstanbul, Türkiye.
Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Yeditepe University, TR-34755, Kayışdağı, İstanbul, Türkiye.
Fitoterapia. 2025 Jul;184:106604. doi: 10.1016/j.fitote.2025.106604. Epub 2025 May 7.
Valeriana tuberosa L. yielded four new iridoids, valtuberoside I-IV (1-3 and 15), along with 13 known secondary metabolites via activity-directed fractionation. Compounds were characterized by NMR and HRESIMS. EtOH extract, fractions, and isolates were evaluated for their inhibition on nitric oxide (NO) release in LPS-induced RAW 264.7 cells. Compounds 3, 4, 6, 8, 9, 11, 13, 16, and 17 exhibited anti-inflammatory activity by inhibiting the release of NO (IC 43.44-95.71 μM), and their mode of actions were elucidated by ELISA, Western blot, qPCR, immunostaining techniques and supported by molecular modelling studies. Compounds 8, 9, 11, 13, and 17 showed significant reduction in TNF-α, IL-1β, IL-6, PGE, and COX-2 enzyme production, while 9 and 13 decreased iNOS protein expression in RAW 264.7 cells. Compound 13 exhibited remarkable inhibition on pro-inflammatory markers, cox-2 gene expression and translocation of NF-κB to the nuclear region. Moreover, it had the most favourable interaction (ds: -6.46 kcal/mol) with iNOS in in silico analyses. The cytotoxic activities of the most active isolates against MCF-7, MDA-MB-231, U87, A172, MIA PaCa-2, PANC-1, Mahlavu, and Hep3B cancer cell lines were assessed using CCK8 assay and their cell death mechanisms were unveiled via Apoptosis/Necrosis Assay Kit. Compound 8 had significant cytotoxic activity against MIA PaCa-2 (IC 23.7 μM) and Hep3B (IC 25.4 μM) cancer cell lines, via arresting cell cycle especially in G2/M phase and triggering the apoptotic pathway. These findings indicated that 8 and 13 deserve further in vivo assays on the way to discover new potential drug leads.
通过活性导向分离，块茎缬草（Valeriana tuberosa L.）产生了四种新的环烯醚萜类化合物，缬草块茎苷I-IV（1-3和15），以及13种已知的次生代谢产物。化合物通过核磁共振（NMR）和高分辨电喷雾电离质谱（HRESIMS）进行表征。对乙醇提取物、馏分和分离物进行评估，以考察它们对脂多糖（LPS）诱导的RAW 264.7细胞中一氧化氮（NO）释放的抑制作用。化合物3、4、6、8、9、11、13、16和17通过抑制NO释放（IC50为43.44-95.71 μM）表现出抗炎活性，并通过酶联免疫吸附测定（ELISA）、蛋白质免疫印迹法（Western blot）、实时荧光定量聚合酶链反应（qPCR）、免疫染色技术阐明其作用模式，并得到分子模拟研究的支持。化合物8、9、11、13和17使RAW 264.7细胞中肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、前列腺素E（PGE）和环氧化酶-2（COX-2）酶的产生显著减少，而化合物9和13降低了RAW 264.7细胞中诱导型一氧化氮合酶（iNOS）蛋白的表达。化合物13对促炎标志物、COX-2基因表达以及核因子-κB（NF-κB）向核区域的转位表现出显著抑制作用。此外，在计算机模拟分析中，它与iNOS具有最有利的相互作用（结合能：-6.46 kcal/mol）。使用细胞计数试剂盒（CCK8）评估活性最高的分离物对MCF-7、MDA-MB-231、U87、A172、MIA PaCa-2、PANC-1、Mahlavu和Hep3B癌细胞系的细胞毒性活性，并通过凋亡/坏死检测试剂盒揭示其细胞死亡机制。化合物8对MIA PaCa-2（IC50为23.7 μM）和Hep3B（IC50为25.4 μM）癌细胞系具有显著的细胞毒性活性，通过使细胞周期停滞，尤其是在G2/M期并触发凋亡途径。这些发现表明，化合物8和13在发现新的潜在药物先导物的道路上值得进一步进行体内试验。
