Insights into Clematis cirrhosa L. Ethanol Extract: Cytotoxic Effects, LC-ESI-QTOF-MS/MS Chemical Profiling, Molecular Docking, and Acute Toxicity Study.

IF 4.3 3区医学 Q2 CHEMISTRY, MEDICINAL

Pharmaceuticals Pub Date : 2024-10-09 DOI:10.3390/ph17101347

Manal I Alruwad, Riham Salah El Dine, Abdallah M Gendy, Abdulrahman M Saleh, Mohamed A Khalaf, Hala M El Hefnawy, Manal M Sabry

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引用次数: 0

Abstract

Background: In Jordanian traditional medicine, Clematis cirrhosa is commonly employed for the management of different diseases. Numerous investigations have documented the cytotoxic properties of different Clematis species against numerous types of cancer. Previously, we demonstrated the potential cytotoxicity of Clematis cirrhosa against HT-29 colorectal cancer cells. Extending our work, the current research aimed to explore the possible mechanisms underlying its antiproliferative activity with a plant safety evaluation.

Methods: This study evaluates the extract's impact on the cell cycle, apoptosis, and cell migration through in vitro assays, LC-ESI-QTOF-MS/MS analysis, docking studies, and an acute toxicity evaluation.

Results: The Clematis cirrhosa ethanol extract (CEE) induced G2/M phase cell cycle arrest (19.63%), triggered significant apoptosis (41.99%), and inhibited cell migration/wound healing by 28.15%. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis revealed increased expression of the proapoptotic markers BAX (6.03-fold) and caspase-3 (6.59-fold), along with the reduced expression of the antiapoptotic BCL-2, in CEE-treated cells. Moreover, CEE significantly restrained angiogenesis by reducing VEGF mRNA expression by 63.9%. High-resolution LC-ESI-QTOF-MS/MS studies identified 26 metabolites, including phenolic compounds, fatty acids, and triterpenoids. Docking studies suggested that manghaslin had the highest binding affinity for VEGFR-2, followed by calceolarioside B, quercetin 7-O-rhamnopyranoside, luteolin, and quercetin-3,7-O-diglucoside. On the other hand, salvadoraside exhibited the highest binding affinity for the inhibition of caspase-3, followed by quercetin-3,7-O-diglucoside, kaempferol-3,7-O-α-L-dirhamnoside, manghaslin, and tectoridin, supporting the observed apoptotic effects. Interestingly, the outcomes further indicate that a single oral administration of up to 5000 mg/kg CEE is safe for consumption.

Conclusions: These outcomes point to the potential of Clematis cirrhosa as a promising candidate for further exploration in cancer therapy.

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对铁线莲乙醇提取物的深入研究：细胞毒性作用、LC-ESI-QTOF-MS/MS 化学成分分析、分子对接和急性毒性研究。

背景：在约旦传统医学中，铁线莲（Clematis cirrhosa）常用于治疗各种疾病。大量研究表明，不同种类的铁线莲对多种癌症具有细胞毒性。此前，我们证明了铁线莲（Clematis cirrhosa）对 HT-29 大肠癌细胞的潜在细胞毒性。目前的研究旨在扩展我们的工作，探索其抗增生活性的可能机制，并进行植物安全性评估：本研究通过体外实验、LC-ESI-QTOF-MS/MS 分析、对接研究和急性毒性评估，评估了提取物对细胞周期、细胞凋亡和细胞迁移的影响：结果：铁线莲乙醇提取物（CEE）能诱导 G2/M 期细胞周期停滞（19.63%），引发细胞显著凋亡（41.99%），抑制细胞迁移/伤口愈合（28.15%）。定量反转录聚合酶链反应（qRT-PCR）分析表明，在 CEE 处理的细胞中，促凋亡标志物 BAX（6.03 倍）和 caspase-3 （6.59 倍）的表达增加，而抗凋亡标志物 BCL-2 的表达减少。此外，CEE 还能减少 63.9% 的血管内皮生长因子 mRNA 表达，从而明显抑制血管生成。高分辨率 LC-ESI-QTOF-MS/MS 研究确定了 26 种代谢物，包括酚类化合物、脂肪酸和三萜类化合物。对接研究表明，曼哈斯林与血管内皮生长因子受体-2的结合亲和力最高，其次是钙黄苷 B、槲皮素 7-O-吡喃鼠李糖苷、木犀草素和槲皮素-3,7-O-二葡萄糖苷。另一方面，沙瓦多苷在抑制 caspase-3 方面表现出最高的结合亲和力，其次是槲皮素-3,7-O-二葡萄糖苷、山奈酚-3,7-O-α-L-二鼠李糖苷、芒柄花苷和橘皮苷，这支持了所观察到的凋亡效应。有趣的是，研究结果进一步表明，单次口服高达 5000 毫克/千克的 CEE 是安全的：结论：这些结果表明，铁线莲有潜力成为癌症治疗领域中值得进一步探索的候选药物。

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来源期刊

Pharmaceuticals Pharmacology, Toxicology and Pharmaceutics-Pharmaceutical Science

CiteScore

6.10

自引率

4.30%

发文量

1332

审稿时长

6 weeks

期刊介绍： Pharmaceuticals (ISSN 1424-8247) is an international scientific journal of medicinal chemistry and related drug sciences.Our aim is to publish updated reviews as well as research articles with comprehensive theoretical and experimental details. Short communications are also accepted; therefore, there is no restriction on the maximum length of the papers.