Laboratorio de Química de Productos Naturales, Instituto de Química de Recursos Naturales, Universidad de Talca, Campus Lircay, Talca, 3460000, Chile.
Laboratorio de Química de Productos Naturales, Instituto de Química de Recursos Naturales, Universidad de Talca, Campus Lircay, Talca, 3460000, Chile; Universidad Nacional de Asunción, Facultad de Ciencias Químicas, Campus San Lorenzo, P.O. Box 1055, Paraguay.
J Ethnopharmacol. 2022 Aug 10;294:115344. doi: 10.1016/j.jep.2022.115344. Epub 2022 May 6.
In Paraguay, healers from the Mbya culture treat cancer with a recipe prepared with the native toad Rhinella schneideri. However, the chemical composition and biological effects of the recipe remain unknown.
The aim is to determine the composition of the traditional preparation made using the toad R. schneideri and to evaluate its effect on human breast cancer (BC) cells.
The metabolites contained in the preparation were concentrated using XAD-7 resin, and the concentrate was analyzed by HPLC-MS/MS. The effect of the preparation was assessed in normal (MCF10F) and BC cells (MDA-MB-231 and MCF7). The mitochondrial membrane potential (Δψm), reactive oxygen species (ROS) levels, and cell cycle progression were determined by flow cytometry. The oxygen consumption rate (OCR) was measured by Clark electrode, and fibronectin-dependent migration in normoxia and hypoxia-like conditions were evaluated by transwell assay.
From the Amberlite-retained extract from the preparation, 24 compounds were identified, including alkaloids, amino acids, bufadienolides, and flavonoids, among others. The crude extract (CE) did not affect cell cycle progression and viability of BC cell lines. Moreover, it did not make cancer cells more sensitive to the cytotoxic effect of the chemotherapeutics doxorubicin and teniposide. On the other hand, the CE reduced the menadione-induced ROS production and increased NADH, Δψm, and the OCR. Respiratory complexes I and III as well as ATP synthase levels were increased in an AMPK-dependent manner. Moreover, the CE inhibited the migration of BC cells in normoxia and a hypoxia-like condition using CoCl as a HIF1α-stabilizing agent. This latter effect involved an AMPK-dependent reduction of HIF1α levels.
The Paraguayan toad recipe contains metabolites from the toad ingredient, including alkaloids and bufadienolide derivatives. The CE lacks cytotoxic effects alone or in combination with chemotherapeutics. However, it increases mitochondrial bioenergetics and inhibits the cancer cell migration in an AMPK-dependent manner in BC cells. This is the first report of the in vitro anticancer effect of a traditional Rhinella sp. toad preparation based on Mbya tradition.
在巴拉圭，米比亚文化的治疗师使用一种用本地蟾蜍 Rhinella schneideri 制备的配方来治疗癌症。然而，该配方的化学成分和生物学效应仍不清楚。
目的是确定使用蟾蜍 R. schneideri 制备的传统制剂的成分，并评估其对人乳腺癌（BC）细胞的作用。
使用 XAD-7 树脂浓缩制剂中的代谢物，并用 HPLC-MS/MS 分析浓缩物。评估制剂对正常（MCF10F）和 BC 细胞（MDA-MB-231 和 MCF7）的作用。通过流式细胞术测定线粒体膜电位（Δψm）、活性氧（ROS）水平和细胞周期进程。通过克拉克电极测量耗氧量（OCR），并通过 Transwell 测定法评估纤连蛋白依赖性在常氧和低氧样条件下的迁移。
从制剂中保留的 Amberlite 提取物中鉴定出 24 种化合物，包括生物碱、氨基酸、蟾蜍毒和黄酮类化合物等。粗提取物（CE）不会影响 BC 细胞系的细胞周期进程和活力。此外，它不会使癌细胞对多柔比星和顺铂等化疗药物的细胞毒性作用更敏感。另一方面，CE 减少了 menadione 诱导的 ROS 产生，并增加了 NADH、Δψm 和 OCR。AMPK 依赖性地增加呼吸复合物 I 和 III 以及 ATP 合酶的水平。此外，CE 抑制了使用 CoCl 作为 HIF1α 稳定化剂的常氧和低氧样条件下 BC 细胞的迁移。后一种作用涉及 AMPK 依赖性的 HIF1α 水平降低。
巴拉圭蟾蜍配方含有来自蟾蜍成分的代谢物，包括生物碱和蟾蜍毒衍生。CE 单独或与化疗药物联合使用时没有细胞毒性作用。然而，它增加了线粒体生物能量学，并以 AMPK 依赖性的方式抑制了 BC 细胞的癌细胞迁移。这是基于 Mbya 传统的传统 Rhinella sp.蟾蜍制剂的体外抗癌作用的首次报道。
