Hu Stephen Chu-Sung, Lee I-Ta, Yen Ming-Hong, Lin Chun-Ching, Lee Chiang-Wen, Yen Feng-Lin
Department of Dermatology, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan; Department of Dermatology, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan.
School of Medicine, College of Medicine, China Medical University, Taichung, Taiwan.
J Ethnopharmacol. 2016 Feb 17;179:432-42. doi: 10.1016/j.jep.2015.12.058. Epub 2015 Dec 31.
Bupleurum chinense is a traditional Chinese medicinal herb which has been used to treat various inflammatory and infectious diseases, while Bupleurum kaoi is an endemic plant in Taiwan. We determined whether B. chinense and B. kaoi and their biologically active saikosaponin compounds possess anti-melanoma activity. In addition, we developed a novel saikosaponin-d nanoparticle system to improve its solubility, and evaluated its antiproliferative effects and molecular mechanisms in melanoma cells.
Ethanolic extracts from B. chinense and B. kaoi were prepared, and their saikosaponin contents were determined by high performance liquid chromatography analysis. Saikosaponin-d nanoparticles were synthesized, and their physicochemical properties were evaluated by particle size analyzer, transmission electron microscopy, differential scanning calorimetry, X-ray diffractometry, and Fourier transform infrared spectroscopy. Human A375.S2 melanoma cells were cultured, and cell viability determined by the MTT assay. Apoptosis was evaluated by determination of mitochondrial membrane potential, and signal transduction pathways investigated by Western blotting.
Ethanolic extracts from B. kaoi showed more potent antiproliferative effect on human A375.S2 melanoma cells compared to B. chinense. The saikosaponin-a, -c and -d contents were higher in B. kaoi compared to B. chinense. Saikosaponin-d was the most potent compound in terms of anti-melanoma activity, and saikosaponin-d nanoparticles exhibited increased water solubility due to lowered particle size, amorphous transformation and intermolecular hydrogen bond formation with the excipient. Furthermore, saikosaponin-d nanoparticles showed enhanced antiproliferative activity against melanoma cells, and induced apoptosis through the mitochondrial pathway. The anti-melanoma activity was mediated by phosphorylation of JNK and p38, phosphorylation of p53, increased level of cytochrome c, and activation of caspase 9.
B. kaoi contains higher saikosaponin content and shows greater anti-melanoma activity than B. chinense. Saikosaponin-d nanoparticles have improved solubility, and may have potential use in the future as a form of treatment for melanoma.
柴胡是一种传统的中草药，已被用于治疗各种炎症和感染性疾病，而台湾柴胡是台湾的一种特有植物。我们确定了柴胡和台湾柴胡及其具有生物活性的柴胡皂苷化合物是否具有抗黑色素瘤活性。此外，我们开发了一种新型的柴胡皂苷 - d纳米颗粒系统以提高其溶解度，并评估了其对黑色素瘤细胞的抗增殖作用和分子机制。
制备了柴胡和台湾柴胡的乙醇提取物，并通过高效液相色谱分析测定了它们的柴胡皂苷含量。合成了柴胡皂苷 - d纳米颗粒，并通过粒度分析仪、透射电子显微镜、差示扫描量热法、X射线衍射法和傅里叶变换红外光谱法评估了其物理化学性质。培养人A375.S2黑色素瘤细胞，并通过MTT法测定细胞活力。通过测定线粒体膜电位评估细胞凋亡，并通过蛋白质印迹法研究信号转导途径。
与柴胡相比，台湾柴胡的乙醇提取物对人A375.S2黑色素瘤细胞显示出更强的抗增殖作用。台湾柴胡中的柴胡皂苷 - a、-c和 - d含量高于柴胡。就抗黑色素瘤活性而言，柴胡皂苷 - d是最有效的化合物，并且柴胡皂苷 - d纳米颗粒由于粒径减小、无定形转变以及与辅料形成分子间氢键而表现出增加的水溶性。此外，柴胡皂苷 - d纳米颗粒对黑色素瘤细胞显示出增强的抗增殖活性，并通过线粒体途径诱导细胞凋亡。抗黑色素瘤活性是由JNK和p38的磷酸化、p53的磷酸化、细胞色素c水平的升高以及caspase 9的激活介导的。
台湾柴胡比柴胡含有更高的柴胡皂苷含量，并且显示出更强的抗黑色素瘤活性。柴胡皂苷 - d纳米颗粒具有改善的溶解度，并且未来可能有潜力作为黑色素瘤的一种治疗形式。
