Huang Weihuan, Zheng Nianjue, Niu Naxin, Tan Ying, Li Yaolan, Tian Haiyan
Key Laboratory of Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou, China; Department of Developmental & Regenerative Biology, Jinan University, Guangzhou, China.
Key Laboratory of Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou, China; Department of Developmental & Regenerative Biology, Jinan University, Guangzhou, China.
J Ethnopharmacol. 2024 Apr 24;324:117811. doi: 10.1016/j.jep.2024.117811. Epub 2024 Jan 28.
Traditionally, the roots of Kaempferia galanga has been used to treat high blood pressure, chest pain, headache, toothache, rheumatism, indigestion, cough, inflammation and cancer in Asia. Nevertheless, most of its pharmacological studies were focused on ethanolic extracts and volatile oils. The exact active chemical constituents and their underlying mechanisms are still poorly understood, especially towards its anti-cancer treatment. Inhibition of angiogenesis is an important atrategy to inhibit tumor growth. It has been reported that the low polar component of the plant possessed anti-angiogenic activity. Yet, the potent compound which is responsible for the effect and its molecular mechanism has not been reported.
To determine the potent anti-angiogenic component in K.galanga and its mechanism of action.
The low polar components of the plant were concentrated using the methods of supercritical fluid extraction (SFE), subcritical extraction (SCE) and steam distillation (SD). The anti-angiogenic activity of the three extracts was evaluated using a zebrafish model. The content of the active compound in those extracts was determined with HPLC analysis. The in-vitro and in-vivo activity of the isolated compound was evaluated using human umbilical vein endothelial cells (HUVECs) model, the aortic ring assay and the matrigel plug assay, respectively. Its molecular mechanism was further studied by the western blotting assay and computer-docking experiments. Besides, its cytotoxicity on cancer and normal cell lines was evaluated using the cell-counting kit.
HPLC results showed that trans-ethyl p-methoxycinnamate (TEM) was the major component of the extracts. The extract of SFE showed the best effect as it has the highest content of TEM. TEM could inhibit vascular endothelial growth factor (VEGF)-induced viability, migration, invasion and tube formation in human umbilical vein endothelial cells (HUVECs) in vitro. Moreover, it inhibited VEGF-induced sprout formation ex vivo and vessel formation in vivo. Mechanistic study showed that it could suppress tyrosine kinase activity of the receptor of VEGF (VEGFR2) and alter its downstream signaling pathways. In addition, the molecular docking showed that the binding of TEM and VEGFR2 is stable, which mainly attributed to the non-covalent binding interaction. Beside, TEM possessed little toxicity to both cancer and normal cells.
TEM is the major anti-angiogenic component present in K. galanga and its anti-angiogenic property rather than toxicity provides scientific basis for the traditional use of K. galanga in cancer treatment.
传统上，在亚洲，山柰的根被用于治疗高血压、胸痛、头痛、牙痛、风湿病、消化不良、咳嗽、炎症和癌症。然而，其大多数药理学研究都集中在乙醇提取物和挥发油上。其确切的活性化学成分及其潜在机制仍知之甚少，尤其是在其抗癌治疗方面。抑制血管生成是抑制肿瘤生长的重要策略。据报道，该植物的低极性成分具有抗血管生成活性。然而，负责该作用的有效化合物及其分子机制尚未见报道。
确定山柰中有效的抗血管生成成分及其作用机制。
采用超临界流体萃取（SFE）、亚临界萃取（SCE）和水蒸气蒸馏（SD）方法浓缩该植物的低极性成分。使用斑马鱼模型评估这三种提取物的抗血管生成活性。通过高效液相色谱（HPLC）分析测定这些提取物中活性化合物的含量。分别使用人脐静脉内皮细胞（HUVECs）模型、主动脉环试验和基质胶栓试验评估分离出的化合物的体外和体内活性。通过蛋白质印迹分析和计算机对接实验进一步研究其分子机制。此外，使用细胞计数试剂盒评估其对癌细胞系和正常细胞系的细胞毒性。
HPLC结果表明，对甲氧基肉桂酸乙酯（TEM）是提取物中的主要成分。SFE提取物效果最佳，因为其TEM含量最高。TEM在体外可抑制血管内皮生长因子（VEGF）诱导的人脐静脉内皮细胞（HUVECs）的活力、迁移、侵袭和管腔形成。此外，它在体外抑制VEGF诱导的芽生形成，在体内抑制血管形成。机制研究表明，它可抑制VEGF受体（VEGFR2）的酪氨酸激酶活性并改变其下游信号通路。此外，分子对接表明TEM与VEGFR2的结合是稳定的，这主要归因于非共价结合相互作用。此外，TEM对癌细胞和正常细胞的毒性都很小。
TEM是山柰中主要的抗血管生成成分，其抗血管生成特性而非毒性为山柰在癌症治疗中的传统应用提供了科学依据。
