HPLC-ESI/MS-MS 分析多穗柯提取物中的化合物并评价其细胞毒性和抗病毒活性：诱导乳腺癌细胞凋亡的作用机制。
HPLC-ESI/MS-MS characterization of compounds in Dolomiaea costus extract and evaluation of cytotoxic and antiviral properties: molecular mechanisms underlying apoptosis-inducing effect on breast cancer.
机构信息
Department of Pharmacognosy, Faculty of Pharmacy, Ain Shams University, Abbassia, 11566, Cairo, Egypt.
Centre of Drug Discovery Research and Development, Ain Shams University, Cairo, Egypt.
出版信息
BMC Complement Med Ther. 2023 Oct 6;23(1):354. doi: 10.1186/s12906-023-04164-9.
BACKGROUND
Dolomiaea costus (syn: Saussurea costus; Family Asteraceae) occupies an important place in the traditional Chinese medicinal plants and is prescribed for a wide range of disorders. The current study aimed to tentatively identify the phytoconstituents of D. costus extract and to explore antiproliferative activity against human breast cancer cells and its possible apoptotic mechanism along with antiviral activity against human adenovirus 5 (Adv-5).
METHODS
The phytoconstituents of 70% ethanol extract of D. costus were assessed using HPLC/ESI-MS/MS technique. The cell viability was investigated against breast cancer cell line (MCF-7) via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Mechanistically, the apoptotic effects on the Bax, Bcl2 and Caspase 3 were determined via quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). Further, the antiviral activity was assessed against Adv-5 based on virucidal and adsorption mechanisms.
RESULTS
The HPLC/MS analysis of the extract revealed tentative identification of twenty compounds of polyphenolic nature, mainly flavonoids, lignans, coumarins, and anthocyanidins. The plant extract showed a cytotoxic effect against MCF-7 and Vero cells with IC values of 15.50 and 44 µg/ml, respectively, indicating its aggressiveness against the proliferation of breast cancer cells as confirmed by apoptotic genes expression which revealed upregulation of Bax and Caspase 3 but further insight analysis is needed to explore exact mechanistic pathway. Antiviral activity against Adv-5 was observed at a non-toxic concentration of the tested extract.
CONCLUSIONS
Such observations against human breast cancer and viral replication supported further studies for nanoformulations in drug delivery systems as targeting therapy and in vivo studies before biomedical applications.
背景
蓝盆花（同义词：Saussurea costus；菊科）在传统中药中占有重要地位，用于治疗多种疾病。本研究旨在初步鉴定蓝盆花提取物的植物成分，并探讨其对人乳腺癌细胞的增殖抑制活性及其可能的凋亡机制，以及对人腺病毒 5（Adv-5）的抗病毒活性。
方法
采用高效液相色谱/电喷雾串联质谱（HPLC/ESI-MS/MS）技术对蓝盆花 70%乙醇提取物的植物成分进行评估。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法检测对乳腺癌细胞系（MCF-7）的细胞活力。通过定量逆转录聚合酶链反应（RT-qPCR）测定凋亡相关蛋白 Bax、Bcl2 和 Caspase 3 的表达，进一步研究其凋亡作用。此外，基于病毒灭活和吸附机制，评估该提取物对 Adv-5 的抗病毒活性。
结果
提取物的 HPLC/MS 分析结果初步鉴定了 20 种多酚类化合物，主要为类黄酮、木脂素、香豆素和花色苷。该植物提取物对 MCF-7 和 Vero 细胞表现出细胞毒性，IC50 值分别为 15.50 和 44µg/ml，表明其对乳腺癌细胞增殖具有侵袭性，这一点通过凋亡基因表达得到证实，结果显示 Bax 和 Caspase 3 的表达上调，但需要进一步的深入分析来探索确切的机制途径。在非毒性浓度的测试提取物中观察到对 Adv-5 的抗病毒活性。
结论
针对人乳腺癌和病毒复制的这些观察结果支持进一步研究针对纳米制剂的药物传递系统，作为靶向治疗，并在生物医学应用之前进行体内研究。
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