解偶联剂 BAM15 对刚地弓形虫 RH 株和普鲁氏滋养体株的作用。

Effect of the mitochondrial uncoupling agent BAM15 against the Toxoplasma gondii RH strain and Prugniaud strain.

机构信息

Health Science Center, Ningbo University, Ningbo, Zhengjiang Province, 315211, People's Republic of China.

Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Lanzhou, Gansu Province, 730050, People's Republic of China.

出版信息

Parasit Vectors. 2024 Mar 1;17(1):96. doi: 10.1186/s13071-024-06187-8.

DOI:10.1186/s13071-024-06187-8

PMID:38424591

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10905885/

Abstract

BACKGROUND

Toxoplasmosis is a zoonotic disease caused by the infection of the protozoa Toxoplasma gondii (T. gondii), and safe and effective therapeutic drugs are lacking. Mitochondria, is an important organelle that maintains T. gondii survival, however, drugs targeting mitochondria are lacking.

METHODS

The cytotoxicity of BAM15 was detected by CCK-8 and the in vitro effects of BAM15 was detected by qPCR, plaque assay and flow cytometry. Furthermore, the ultrastructural changes of T. gondii after BAM15 treatment were observed by transmission electron microscopy, and further the mitochondrial membrane potential (ΔΨm), ATP level and reactive oxygen species (ROS) of T. gondii after BAM15 treatment were detected. The pharmacokinetic experiments and in vivo infection assays were performed in mice to determine the in vivo effect of BAM15.

RESULTS

BAM15 had excellent anti-T. gondii activity in vitro and in vivo with an EC50 value of 1.25 μM, while the IC50 of BAM15 in Vero cells was 27.07 μM. Notably, BAM15 significantly inhibited proliferation activity of T. gondii RH strain and Prugniaud strain (PRU), caused T. gondii death. Furthermore, BAM15 treatment induced T. gondii mitochondrial vacuolation and autolysis by TEM. Moreover, the decrease in ΔΨm and ATP level, as well as the increase in ROS production further confirmed the changes CONCLUSIONS: Our study identifies a useful T. gondii mitochondrial inhibitor, which may also serve as a leading molecule to develop therapeutic mitochondrial inhibitors in toxoplasmosis.'

摘要

背景

弓形虫病是一种由原生动物弓形虫（Toxoplasma gondii，T. gondii）感染引起的人畜共患疾病，目前缺乏安全有效的治疗药物。线粒体是维持弓形虫生存的重要细胞器，但缺乏针对线粒体的药物。

方法

用 CCK-8 检测 BAM15 的细胞毒性，用 qPCR、噬菌斑试验和流式细胞术检测 BAM15 的体外作用。进一步用透射电子显微镜观察 BAM15 处理后弓形虫的超微结构变化，进一步检测 BAM15 处理后弓形虫的线粒体膜电位（ΔΨm）、ATP 水平和活性氧（ROS）。在小鼠中进行药代动力学实验和体内感染实验，以确定 BAM15 的体内作用。

结果

BAM15 具有良好的抗弓形虫活性，体外和体内的 EC50 值分别为 1.25 μM 和 2.70 μM，而 BAM15 在 Vero 细胞中的 IC50 值为 27.07 μM。值得注意的是，BAM15 显著抑制 RH 株和 Prugniaud 株（PRU）弓形虫的增殖活性，导致弓形虫死亡。此外，BAM15 处理通过 TEM 诱导弓形虫线粒体空泡化和自溶。此外，ΔΨm 和 ATP 水平的降低以及 ROS 产生的增加进一步证实了这些变化。