从龙葵中分离得到的生物活性甾体生物碱甾体生物碱苦茄碱可诱导非选择性细胞毒性和 P 糖蛋白抑制。
Solamargine, a bioactive steroidal alkaloid isolated from Solanum aculeastrum induces non-selective cytotoxicity and P-glycoprotein inhibition.
机构信息
Department of Pharmacology, School of Medicine, Faculty of Health Sciences, University of Pretoria, P.O.Box X323, Pretoria, Arcadia, 0007, South Africa.
Biosciences Division, Council of Scientific and Industrial Research, Pretoria, South Africa.
出版信息
BMC Complement Altern Med. 2018 May 2;18(1):137. doi: 10.1186/s12906-018-2208-7.
BACKGROUND
Solanum aculeastrum fruits are used by some cancer sufferers as a form of alternative treatment. Scientific literature is scarce concerning its anticancer activity, and thus the aim of the study was to assess the in vitro anticancer and P-glycoprotein inhibitory potential of extracts of S. aculeastrum fruits. Furthermore, assessment of the combinational effect with doxorubicin was also done.
METHODS
The crude extract was prepared by ultrasonic maceration. Liquid-liquid extraction yielded one aqueous and two organic fractions. Bioactive constituents were isolated from the aqueous fraction by means of column chromatography, solid phase extraction and preparative thin-layer chromatography. Confirmation of bioactive constituent identity was done by nuclear magnetic resonance and ultra-performance liquid chromatography mass spectrometry. The crude extract and fractions were assessed for cytotoxicity and P-glycoprotein inhibition in both cancerous and non-cancerous cell lines using the sulforhodamine B and rhodamine-123 assays, respectively.
RESULTS
Both the crude extract and aqueous fraction was cytotoxic to all cell lines, with the SH-SY5Y neuroblastoma cell line being most susceptible to exposure (IC = 10.72 μg/mL [crude], 17.21 μg/mL [aqueous]). Dose-dependent P-glycoprotein inhibition was observed for the crude extract (5.9 to 18.9-fold at 100 μg/mL) and aqueous fraction (2.9 to 21.2 at 100 μg/mL). The steroidal alkaloids solamargine and solanine were identified. While solanine was not bioactive, solamargine displayed an IC of 15.62 μg/mL, and 9.1-fold P-glycoprotein inhibition at 100 μg/mL against the SH-SY5Y cell line. Additive effects were noted for combinations of doxorubicin against the SH-SY5Y cell line.
CONCLUSIONS
The crude extract and aqueous fraction displayed potent non-selective cytotoxicity and noteworthy P-glycoprotein inhibition. These effects were attributed to solamargine. P-glycoprotein inhibitory activity was only present at concentrations higher than those inducing cytotoxicity, and thus does not appear to be the likely mechanism for the enhancement of doxorubicin's cytotoxicity. Preliminary results suggest that non-selective cytotoxicity may hinder drug development, however, further assessment of the mode of cell death is necessary to determine the route forward.
背景
龙葵果被一些癌症患者用作替代疗法。关于其抗癌活性的科学文献很少，因此本研究旨在评估龙葵果提取物的体外抗癌和 P-糖蛋白抑制潜力。此外，还评估了与阿霉素的联合作用。
方法
通过超声浸提制备粗提物。液液萃取得到一个水相和两个有机相。通过柱色谱、固相萃取和制备薄层层析从水相分离出生物活性成分。通过核磁共振和超高效液相色谱质谱联用确认生物活性成分的身份。通过磺基罗丹明 B 和罗丹明 123 测定法分别在癌细胞和非癌细胞系中评估粗提取物和馏分的细胞毒性和 P-糖蛋白抑制作用。
结果
粗提取物和水相均对所有细胞系具有细胞毒性，其中 SH-SY5Y 神经母细胞瘤细胞系对暴露最敏感（IC=10.72μg/mL[粗提物]，17.21μg/mL[水相]）。粗提取物（在 100μg/mL 时为 5.9 至 18.9 倍）和水相（在 100μg/mL 时为 2.9 至 21.2 倍）显示出剂量依赖性的 P-糖蛋白抑制作用。鉴定出甾体生物碱龙葵碱和茄碱。虽然茄碱没有生物活性，但龙葵碱在 100μg/mL 时对 SH-SY5Y 细胞系的 IC 为 15.62μg/mL，P-糖蛋白抑制作用为 9.1 倍。在 SH-SY5Y 细胞系中，阿霉素联合使用时观察到相加作用。
结论
粗提取物和水相显示出强大的非选择性细胞毒性和显著的 P-糖蛋白抑制作用。这些作用归因于龙葵碱。P-糖蛋白抑制活性仅在诱导细胞毒性的浓度以上存在，因此似乎不是增强阿霉素细胞毒性的可能机制。初步结果表明，非选择性细胞毒性可能会阻碍药物开发，但是，需要进一步评估细胞死亡模式以确定前进的方向。
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