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Hypericum erectum alcoholic extract inhibits Toxoplasma growth and Entamoeba encystation: an exploratory study on the anti-protozoan potential.
Journal of Natural Medicines ( IF 3.3 ) Pub Date : 2019-11-14 , DOI: 10.1007/s11418-019-01369-6
Noriko Shinjyo 1, 2, 3 , Hideyuki Nakayama 4 , Kanji Ishimaru 5 , Kenji Hikosaka 2 , Fumika Mi-Ichi 1 , Kazumi Norose 2 , Hiroki Yoshida 1
Affiliation  

Hypericum erectum is an important ethnobotanical medicine in East Asian tradition. To explore the anti-parasitic potential of H. erectum, inhibitory effects on the growth of intracellular parasite Toxoplasma and on the encystation of intestinal parasite Entamoeba were examined. The constituents in H. erectum alcoholic extracts and fractions separated by solvent-partitioning were analysed by high resolution LC–MS. Toxoplasma gondii growth inhibition assay was performed using GFP-labelled T. gondii strain PTG-GFP by measuring the fluorescence intensity. Anti-Toxoplasma drug pyrimethamine was used as a positive control. T. gondii-induced immune reaction was assessed by quantitative PCR and fluorescence microscopy, using co-culture of PTG-GFP and monocyte-macrophage cell line Raw264. The inhibitory effect on the encystation of Entamoeba invadens was measured by flow-cytometry, where paromomycin was used as a positive control. H. erectum methanol (MeOH) extract (50 µg/mL) and ethyl acetate (EtOAc) fraction (50 µg/mL) inhibited the growth of T. gondii, while 50%MeOH extract and hydrophilic fractions were ineffective. Co-culture with T. gondii reduced the viability of macrophages, however macrophages were protected in the presence of H. erectum MeOH extract or EtOAc fraction (above 10 µg/mL). The MeOH extract and EtOAc fraction also effectively suppressed the encystation of E. invadens at 1 mg/mL. Hypericine, a major constituent in MeOH extract and EtOAc fraction, inhibited T. gondii growth and E. invadens encystation. Our results demonstrated that H. erectum effectively inhibited Toxoplasma growth and Entamoeba encystation. These activities are partly mediated by hypericin. In addition, it was suggested the extract and fraction may protect innate immune cells from Toxoplasma-induced damages, thereby enhancing parasite clearance. Further investigation is warranted to address the in vivo effectiveness of H. erectum as an anti-protozoal medicine.

中文翻译:

直立金丝桃属植物酒精提取物抑制弓形虫的生长和肠虫的侵染:抗原生动物潜力的探索性研究。

直立金丝桃属植物是东亚传统中重要的民族植物药。要探索的抗寄生虫潜在H. erectum,对细胞内寄生虫的生长抑制作用弓形虫和肠道寄生虫的虫包囊阿米巴进行了检查。在成分H. erectum醇提取物和级分分离由溶剂分区是由高分辨率LC-MS进行分析。通过测量荧光强度,使用GFP标记的弓形虫PTG-GFP进行弓形虫生长抑制测定。抗弓形虫药物乙胺嘧啶用作阳性对照。弓形虫使用PTG-GFP和单核巨噬细胞系Raw264的共培养,通过定量PCR和荧​​光显微镜评估诱导的免疫反应。通过流式细胞术测量对肠线虫侵染的抑制作用,其中巴龙霉素被用作阳性对照。直立人甲醇(MeOH)提取物(50 µg / mL)和乙酸乙酯(EtOAc)馏分(50μg/ mL)抑制了弓形虫的生长,而50%MeOH提取物和亲水性馏分无效。与刚地弓形虫共培养降低了巨噬细胞的活力,但是在直立螺旋体存在下巨噬细胞受到保护MeOH萃取液或EtOAc馏分(10 µg / mL以上)。MeOH提取物和EtOAc馏分也以1 mg / mL的浓度有效抑制入侵大肠杆菌的侵染。金丝桃素是MeOH提取物和EtOAc馏分中的主要成分,可抑制刚地弓形虫的生长和入侵。我们的研究结果表明,H. erectum有效抑制弓形虫的生长和阿米巴虫包囊。这些活动部分由金丝桃素介导。此外,有人提出提取物和级分可能保护先天免疫细胞免受弓形虫-诱导的损害,从而提高了寄生虫清除率。有必要进行进一步的研究以解决直立支原体作为抗原生动物药物的体内效力。
更新日期:2019-11-14
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