Department of Pharmacology, Faculty of Medicine, Erciyes University, 38039 Talas/Kayseri, Türkiye.
Genkök Genome and Stem Cell Center, Erciyes University, 38039 Talas/Kayseri, Türkiye.
Med Mycol. 2024 Aug 2;62(8). doi: 10.1093/mmy/myae072.
Microsporidia are obligate, intracellular, spore-forming eukaryotic fungi that infect humans and animals. In the treatment of disseminated microsporidiosis albendazole is the choice of drug. In recent years, antiparasitic activity of phosphodiesterase (PDE) enzyme inhibitors has been demonstrated against parasites and fungi, however, there is no information on microsporidia. Vinpocetine is currently used as a cerebral vasodilator drug and also as a dietary supplement to improve cognitive functions. Vinpocetine inhibits PDE1, so we aimed to investigate whether vinpocetine alone or in combination with albendazole has any effect on the spore load of Encephalitozoon intestinalis (E. intestinalis)-infected HEK293 cells. After determining the noncytotoxic concentrations of vinpocetine and albendazole on the host cell by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, HEK293 cells were infected with E. intestinalis spores. Then, two different concentrations of vinpocetine, albendazole, and a combination of both drugs were applied to the cells with an interval of 72 h for 15 days. Spore load of the cells was analyzed by real-time PCR. After the last treatment, spore Deoxyribonucleic Acid (DNA) load was significantly reduced only in the group treated with 14 ng/ml albendazole. It was not different from control in groups treated with 7 ng/ml albendazole and 4-20 µM vinpocetine. However, the combination of vinpocetine significantly increased the effect of albendazole at both concentrations. To our knowledge, this is the first study to investigate the microsporicidal activity of vinpocetine as well as its combinations with albendazole. However, further studies are needed to investigate the mechanism of action and also confirm in vivo conditions.
微孢子虫是一种专性、细胞内、孢子形成的真核真菌,可感染人类和动物。在治疗播散性微孢子虫病时,阿苯达唑是首选药物。近年来,磷酸二酯酶(PDE)酶抑制剂的抗寄生虫和抗真菌活性已得到证实,然而,关于微孢子虫的信息尚不清楚。长春西汀目前用作脑血管扩张药物,也用作改善认知功能的膳食补充剂。长春西汀抑制 PDE1,因此我们旨在研究长春西汀单独或与阿苯达唑联合使用是否对感染肠微孢子虫(E. intestinalis)的 HEK293 细胞的孢子负荷有影响。通过 MTT(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴盐)测定法确定长春西汀和阿苯达唑对宿主细胞的非细胞毒性浓度后,HEK293 细胞被肠微孢子虫孢子感染。然后,将两种不同浓度的长春西汀、阿苯达唑和两者的组合以 72 小时的间隔应用于细胞中,持续 15 天。通过实时 PCR 分析细胞的孢子负荷。在最后一次治疗后,仅在用 14ng/ml 阿苯达唑治疗的组中,孢子脱氧核糖核酸(DNA)负荷显著降低。在用 7ng/ml 阿苯达唑和 4-20µM 长春西汀治疗的组中与对照无差异。然而,长春西汀的组合在两种浓度下均显著增强了阿苯达唑的效果。据我们所知,这是第一项研究长春西汀的杀孢子活性及其与阿苯达唑的组合的研究。然而,需要进一步研究以探讨其作用机制,并在体内条件下进行确认。