Gyi Khin Khin, Anuchapreeda Songyot, Intasai Nutjeera, Tungjai Montree, Okonogi Siriporn, Iwasaki Arihiro, Usuki Toyonobu, Tima Singkome
Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai, 50200, Thailand.
Faculty of Associated Medical Sciences, Chiang Mai University, Under The CMU Presidential Scholarship, Chiang Mai, 50200, Thailand.
BMC Complement Med Ther. 2025 May 13;25(1):172. doi: 10.1186/s12906-025-04903-0.
Gynostemma pentaphyllum (Thunb.), a traditional adaptogenic herb, is known for its bioactive components with potential anti-cancer properties. Acute myeloid leukemia (AML) progression is significantly influenced by Feline McDonough Sarcoma (FMS)-like tyrosine kinase 3 (FLT3) signaling, while Wilms' tumor 1 (WT1) serves as a key prognostic marker. This study investigates the anti-leukemia activities of active G. pentaphyllum leaf extracts and their components, focusing on the inhibition of FLT3 and WT1 activity.
G. pentaphyllum extracts were prepared through maceration, yielding three crude fractional extracts. The cytotoxicity of the extracts was screened against various leukemia cell lines using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The most cytotoxic extract was further fractionated and purified via column chromatography. The anti-proliferative and apoptotic induction activities of the active extract and its fraction were evaluated through cell cycle and apoptosis analyses using flow cytometry. Changes in mitochondrial membrane potential (ΔΨm) were assessed by spectrofluorometry. To confirm anti-leukemia activity, the expression levels of FLT3, WT1 and apoptotic-related protein were analyzed using Western blotting. The major active compounds within the active fractions were identified and characterized using Electrospray Ionization Mass Spectrometry (ESI-MS) and Nuclear Magnetic Resonance (NMR) spectroscopy.
The ethyl acetate fractional extract (F-EtOAc) demonstrated the highest cytotoxicity, particularly against FLT3-overexpressing EoL-1 (IC = 40.82 ± 0.8 µg/mL) and MV4-11 (IC = 35.54 ± 4.1 µg/mL) AML cell lines. Fraction F10 was identified as the most active fraction, significantly inhibited FLT3 and WT1 protein expression and induced G0/G1 cell cycle arrest in a dose-dependent manner. Additionally, F10 induced dose-dependent apoptosis through disruption of ΔΨm, p53 up-regulation and caspase-3 activation. Further purification of F10 identified dehydrovomifoliol as its major bioactive compound.
These findings suggest that the ethyl acetate extract of G. pentaphyllum contains bioactive compounds with anti-leukemia potential, warranting further investigation to evaluate its efficacy against AML.
Not applicable.
绞股蓝是一种传统的适应原性草药，以其具有潜在抗癌特性的生物活性成分而闻名。急性髓系白血病（AML）的进展受到猫 McDonough 肉瘤（FMS）样酪氨酸激酶 3（FLT3）信号传导的显著影响，而肾母细胞瘤 1（WT1）是一个关键的预后标志物。本研究调查了绞股蓝叶活性提取物及其成分的抗白血病活性，重点关注对 FLT3 和 WT1 活性的抑制作用。
通过浸渍法制备绞股蓝提取物，得到三种粗分提取物。使用 3 -（4,5 - 二甲基噻唑 - 2 - 基）- 2,5 - 二苯基四氮唑溴盐（MTT）法筛选提取物对各种白血病细胞系的细胞毒性。对细胞毒性最强的提取物通过柱色谱进一步分离和纯化。通过流式细胞术进行细胞周期和凋亡分析，评估活性提取物及其馏分的抗增殖和凋亡诱导活性。通过荧光光谱法评估线粒体膜电位（ΔΨm）的变化。为了确认抗白血病活性，使用蛋白质免疫印迹法分析 FLT3、WT1 和凋亡相关蛋白的表达水平。使用电喷雾电离质谱（ESI - MS）和核磁共振（NMR）光谱对活性馏分中的主要活性化合物进行鉴定和表征。
乙酸乙酯粗分提取物（F - EtOAc）表现出最高的细胞毒性，尤其是对过表达 FLT3 的 EoL - 1（IC = 40.82 ± 0.8 µg/mL）和 MV4 - 11（IC = 35.54 ± 4.1 µg/mL）AML 细胞系。馏分 F10 被确定为最具活性的馏分，显著抑制 FLT3 和 WT1 蛋白表达，并以剂量依赖性方式诱导 G0/G1 细胞周期阻滞。此外，F10 通过破坏 ΔΨm、上调 p53 和激活 caspase - 3 诱导剂量依赖性凋亡。对 F10 的进一步纯化鉴定出脱氢催吐萝芙木醇为其主要生物活性化合物。
这些发现表明绞股蓝的乙酸乙酯提取物含有具有抗白血病潜力的生物活性化合物，值得进一步研究以评估其对 AML 的疗效。
不适用。
