Herrera-Martínez Mayra, Hernández-Ramírez Verónica Ivonne, Montaño Sarita, Chávez-Munguía Bibiana, Hernández-Carlos Beatriz, Talamás-Rohana Patricia
Instituto de Farmacobiología, Universidad de la Cañada, Carretera Teotitlán - San Antonio Nanahuatipán Km 1.7 s/n., Paraje Titlacuatitla, Teotitlán de Flores Magón, Oaxaca 68540, Mexico.
Departmento de Infectómica y Patogénesis Molecular, CINVESTAV-IPN, Av. IPN 2508, San Pedro Zacatenco, Ciudad de México 07360, Mexico.
Mol Biochem Parasitol. 2022 Nov;252:111512. doi: 10.1016/j.molbiopara.2022.111512. Epub 2022 Sep 6.
This study aimed to know if alpha terthienyl (α-T) affects E. histolytica viability and to analyze its effect on the actin cytoskeleton. Trophozoites of E. histolytica HM1-IMSS were treated with α-T, then, cell viability and morphology were evaluated using tetrazolium salts and scanning electron microscopy, respectively; while actin filaments (F-actin) were stained with rhodamine-phalloidin, observed by confocal microscopy and quantified by fluorometry. Data showed that α-T inhibited cell viability of trophozoites (IC, 19.43 µg / mL), affected the cell morphology, and increased the F-actin in a dose-dependent manner. Production of reactive oxygen species and RhoA-GTP levels remained normal in α-T-treated amebas. Two inhibitors that affect the organization of the trophozoites cytoskeleton, one that interacts directly with actin, Cytochalasin D (CD), and one that affects the Rho signaling pathway by inhibiting the downstream effector Rock, Y27632, were tested. Y27632 did not affect the increase of polymerized actin observed with α-T, this compound partially ameliorates the potent disrupting effects of CD on actin filaments. Docking results suggest that α-T could be an antagonist of CD for the same interaction zone in actin, however, more studies are needed to define the action mechanism of this compound.
本研究旨在了解α-三联噻吩(α-T)是否影响溶组织内阿米巴的活力,并分析其对肌动蛋白细胞骨架的影响。用α-T处理溶组织内阿米巴HM1-IMSS滋养体,然后分别使用四氮唑盐和扫描电子显微镜评估细胞活力和形态;同时用罗丹明-鬼笔环肽对肌动蛋白丝(F-肌动蛋白)进行染色,通过共聚焦显微镜观察并用荧光法进行定量。数据显示,α-T抑制滋养体的细胞活力(IC,19.43µg/mL),影响细胞形态,并以剂量依赖性方式增加F-肌动蛋白。在经α-T处理的变形虫中,活性氧的产生和RhoA-GTP水平保持正常。测试了两种影响滋养体细胞骨架组织的抑制剂,一种直接与肌动蛋白相互作用的细胞松弛素D(CD),另一种通过抑制下游效应器Rock影响Rho信号通路的Y27632。Y27632不影响用α-T观察到的聚合肌动蛋白的增加,该化合物部分改善了CD对肌动蛋白丝的强效破坏作用。对接结果表明,α-T可能是CD在肌动蛋白中相同相互作用区域的拮抗剂,然而,需要更多的研究来确定该化合物的作用机制。