Departamento de Investigación en Alimentos, Facultad de Ciencias Químicas, Universidad Autónoma de Coahuila, Saltillo, Coahuila, Mexico.
Departamento de Química, Centro Universitario de Ciencias Exactas e Ingenierías, Universidad de Guadalajara, Guadalajara, Jalisco, Mexico.
PeerJ. 2022 Apr 27;10:e13350. doi: 10.7717/peerj.13350. eCollection 2022.
Diarrheal diseases caused by protozoa have a great impact on human health around the world. is one of the most common flagellates in the intestinal tract. Factors such as adverse effects to first-line drugs or the appearance of drug-resistant strains, make it necessary to identify new treatment alternatives. Agroindustry waste, like pomegranate peel, are a source of phenolic compounds, which possess antiparasitic activities. studies demonstrated antigiardiasic potential by reducing cyst shedding and protecting intestinal cells; however, they did not identify the compounds or elucidate any mechanism of action in the parasite. The objective of this study is to identify potential molecular targets and to test the effects of polyphenols from on .
The antigiardial potential of polyphenolic extract from pomegranate peel ( L.) obtained using microwave-ultrasound methodology was evaluated on trophozoites. Extract phytochemical identification was performed by HPLC/MS analysis. The effect of polyphenolic extract on growth and adhesion capacity was determined by parasite kinetics; morphological damage was evaluated by SEM, alteration on α-tubulin expression and distribution were analyzed by western blot and immunofluorescence, respectively.
The pomegranate peel extract showed the presence of ellagitannins (punicalin and punicalagin, galloyl-dihexahydroxydiphenoyl-hexoside), flavones (luteolin), and ellagic acid, that caused an inhibitory effect on growth and adhesion capacity, particularly on cells treated with 200 µg/mL, where growth inhibition of 74.36%, trophozoite adherence inhibition of 46.8% and IC of 179 µg/mL at 48 h were demonstrated. The most important findings were that the extract alters α-tubulin expression and distribution in trophozoites in a concentration-independent manner. Also, an increase in α-tubulin expression at 200 µg/mL was observed in western blot and diffuse or incomplete immunolabeling pattern, especially in ventral disk. In addition, the extract caused elongation, disturbance of normal shape, irregularities in the membrane, and flagella abnormalities.
The pomegranate peel extract affects trophozoites . The damage is related to the cytoskeleton, due to expression and distribution alterations in α-tubulin, particularly in the ventral disk, a primordial structure for adhesion and pathogenesis. Microtubule impairment could explain morphological changes, and inhibition of adhesion capacity and growth. Besides, this is the first report that suggests that ellagic acid, punicalin, punicalagin and luteolin could be interactioning with the rich-tubulin cytoskeleton of . Further investigations are needed in order to elucidate the mechanisms of action of the isolated compounds and propose a potential drug alternative for the giardiasis treatment.
由原生动物引起的腹泻病对全球人类健康有重大影响。贾第虫是肠道中最常见的鞭毛虫之一。由于对一线药物的不良反应或出现耐药株等因素，有必要寻找新的治疗替代方法。石榴皮等农业工业废物是酚类化合物的来源，具有抗寄生虫活性。研究表明，通过减少包囊脱落和保护肠细胞，具有抗贾第虫作用；然而，它们没有确定化合物或阐明寄生虫中任何作用机制。本研究的目的是鉴定潜在的分子靶标，并测试石榴皮多酚对 的作用。
采用微波超声法从石榴皮（L.）中提取多酚，评价其对滋养体的抗贾第虫活性。采用高效液相色谱/质谱分析进行提取物的植物化学鉴定。通过寄生虫动力学测定多酚提取物对生长和黏附能力的影响；通过扫描电子显微镜（SEM）评估形态损伤，通过 Western blot 和免疫荧光分别分析α-微管蛋白表达和分布的改变。
石榴皮提取物中存在鞣花单宁（鞣花酸、安石榴甙）、黄酮类（芦丁）和鞣花酸，对生长和黏附能力有抑制作用，特别是在 200µg/mL 时，生长抑制率为 74.36%，滋养体黏附抑制率为 46.8%，48 h 的 IC为 179µg/mL。最重要的发现是，提取物以浓度非依赖性方式改变滋养体中的α-微管蛋白表达和分布。此外，在 Western blot 中观察到 200µg/mL 时α-微管蛋白表达增加，免疫荧光标记模式呈弥散或不完整，尤其是在腹吸盘。此外，提取物还导致鞭毛伸长、形状异常、膜不规则和鞭毛异常。
石榴皮提取物影响滋养体。损伤与细胞骨架有关，因为α-微管蛋白的表达和分布发生改变，特别是在腹吸盘，这是黏附和发病的原始结构。微管蛋白损伤可以解释形态变化和黏附能力及生长抑制。此外，这是首次报道鞣花酸、安石榴甙、鞣花酸和芦丁可能与 丰富的微管蛋白细胞骨架相互作用。需要进一步研究以阐明分离化合物的作用机制，并提出一种潜在的替代药物治疗贾第虫病。
