Exploring the anti-inflammatory and cytotoxic effects of Valeriana tuberosa L. constituents: Integrating in vitro and in silico studies

IF 2.5 3区医学 Q3 CHEMISTRY, MEDICINAL

Fitoterapia Pub Date : 2025-05-07 DOI:10.1016/j.fitote.2025.106604

Cansel Çelik , Yağmur Özhan , Ceren Öztürk , Zulal Sevgi Dede , Tugce Citoglu , Burcin Gungor , Başak Aru , Enise Ece Gurdal , Wolfgang Sippl , Hande Sipahi , Mehtap Tekşen , Hasan Kırmızıbekmez

{"title":"Exploring the anti-inflammatory and cytotoxic effects of Valeriana tuberosa L. constituents: Integrating in vitro and in silico studies","authors":"Cansel Çelik , Yağmur Özhan , Ceren Öztürk , Zulal Sevgi Dede , Tugce Citoglu , Burcin Gungor , Başak Aru , Enise Ece Gurdal , Wolfgang Sippl , Hande Sipahi , Mehtap Tekşen , Hasan Kırmızıbekmez","doi":"10.1016/j.fitote.2025.106604","DOIUrl":null,"url":null,"abstract":"<div><div>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 (IC50 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, PGE2, 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 (IC50 23.7 μM) and Hep3B (IC50 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.</div></div>","PeriodicalId":12147,"journal":{"name":"Fitoterapia","volume":"184 ","pages":"Article 106604"},"PeriodicalIF":2.5000,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Fitoterapia","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0367326X25002291","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"CHEMISTRY, MEDICINAL","Score":null,"Total":0}

引用次数: 0

Abstract

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 (IC₅₀ 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, PGE₂, 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 (IC₅₀ 23.7 μM) and Hep3B (IC₅₀ 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.

Abstract Image

查看原文本刊更多论文

探索缬草成分的抗炎和细胞毒作用：体外和计算机研究的整合

通过活性定向分离得到四种新的环烯醚萜类化合物，缬草苷I-IV(1-3和15)，以及13种已知的次生代谢产物。化合物通过NMR和HRESIMS进行了表征。研究了EtOH提取物、馏分和分离物对lps诱导的RAW 264.7细胞一氧化氮（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的释放表现出抗炎活性（IC50为43.44 ~ 95.71 μM），并通过ELISA、Western blot、qPCR、免疫染色技术和分子模型研究证实了它们的作用模式。化合物8、9、11、13和17在RAW 264.7细胞中显著降低TNF-α、IL-1β、IL-6、PGE2和COX-2酶的产生，而9和13降低iNOS蛋白的表达。化合物13对促炎标志物、cox-2基因表达和NF-κB向核区易位有显著抑制作用。此外，在硅分析中，它与iNOS具有最有利的相互作用（ds:−6.46 kcal/mol）。CCK8法检测活性最高的分离株对MCF-7、MDA-MB-231、U87、A172、MIA PaCa-2、PANC-1、Mahlavu和Hep3B癌细胞的细胞毒活性，并通过凋亡/坏死测定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）癌细胞具有显著的细胞毒活性，其作用机制是通过阻滞细胞周期（特别是G2/M期）和触发细胞凋亡通路。这些发现表明，在发现新的潜在药物先导物的过程中，8和13值得进一步的体内试验。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Fitoterapia 医学-药学

CiteScore

5.80

自引率

2.90%

发文量

198

审稿时长

1.5 months

期刊介绍： Fitoterapia is a Journal dedicated to medicinal plants and to bioactive natural products of plant origin. It publishes original contributions in seven major areas: 1. Characterization of active ingredients of medicinal plants 2. Development of standardization method for bioactive plant extracts and natural products 3. Identification of bioactivity in plant extracts 4. Identification of targets and mechanism of activity of plant extracts 5. Production and genomic characterization of medicinal plants biomass 6. Chemistry and biochemistry of bioactive natural products of plant origin 7. Critical reviews of the historical, clinical and legal status of medicinal plants, and accounts on topical issues.