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番石榴籽成分中生物活性化合物在口腔-胃部消化过程中的生物利用度及其对幽门螺杆菌的抗菌能力。

Bioaccessibility of bioactive compounds present in Persea americana Mill. seed ingredient during oral-gastric digestion with antibacterial capacity against Helicobacter pylori.

机构信息

Programa de Posgrado en Alimentos del Centro de la República (PROPAC), Research and Graduate Studies in Food Science, School of Chemistry, Universidad Autónoma de Querétaro, Querétaro, Mexico.

Unidad de Investigación en Enfermedades Infecciosas, Área de Genética Bacteriana, Hospital Infantil de México Federico Gómez, Mexico.

出版信息

J Ethnopharmacol. 2024 Sep 15;331:118259. doi: 10.1016/j.jep.2024.118259. Epub 2024 Apr 27.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

In ancient Mexican cultures, the Persea americana Mill seed has been used against gastrointestinal diseases, due to high concentrations of bioactive compounds. According to Traditional Mexican Medicine, P. americana seed aqueous infusion is used against roundworms, intestinal worms, parasites, and gastrointestinal problems, in a dose taken over three or four days. In addition, Mexican Society of Natural History indicates the traditional use of P. americana seed powder as an antiparasitic, and antibacterial. On the other hand, Helicobacter pylori infection is a factor associated with the development of gastric disease, peptic ulcers as well as some types of gastric lymphomas and gastric cancer in humans; in this way is necessary scientific evidence about P. americana seed effect in gastrointestinal disease.

AIM OF THE STUDY

The work aimed to evaluate bioactive compounds bioaccessibility and antimicrobial potential against Helicobacter pylori during oral-gastric digestion in vitro of food ingredient from Persea americana Mill. seed and elucidate the possible action mechanism using in silico tools.

MATERIALS AND METHODS

Initially, P. americana seed oil and aqueous extract of P. americana seed were obtained using ultrasound and maceration respectively, and the food ingredient from P. americana seed was obtained. The samples underwent oral-gastric digestions by the INFOGEST method, to continue identifying and quantifying the bioactive compounds by HPLC-DAD and GC-MS. The anti-Helicobacter pylori activity determination were used fourteen Helicobacter pylori clinical strains and reference strains by Susceptibility testing by Minimal Inhibition Concentration, Kinetics of Growth Inhibition of H. pylori, Urease Inhibitory Kinetic. Finally, to elucidate a possible action mechanism used in silico tools (Software AutoDock 4.2.6 and BioVia Discovery v.19.1.0.1.18287).

RESULTS

The lipophilic fraction of P. americana seed detected oleic acid, linoleic acid, and avocadenofuran compounds, and the phenolic fraction showed the presence of catechin, rutin, ellagic, and chlorogenic acid, among others. Phenolic compounds conformational changes during oral-gastric digestion due to mechanical and acid hydrolysis, while lipophilic compounds showed a 20% increase in the gastric phase. Persea americana Mill. seed ingredient (3.08 μg/mL) showed total in vitro inhibition of clinical and reference strains of H. pylori, likewise, the lipophilic fraction had a lower inhibition concentration (2.59 μg/mL) regardless of the strains. Among the mechanisms found in silico, inhibition of target proteins such as CagA, BabA, and MUC5 were observed, as virulence factors involving adherence and bacterial pathogenicity.

CONCLUSIONS

This research provides evidence that food ingredient from P. americana seed has antimicrobial in vitro potential against H. pylori clinical strains, through phenolic and mainly lipophilic compounds, opening new scientific evidence that supports the P. americana seed's traditional use.

摘要

民族药理学相关性

在古墨西哥文化中,美洲蒲桃的种子被用于治疗胃肠道疾病,因为它含有高浓度的生物活性化合物。根据传统墨西哥医学,美洲蒲桃种子的水浸液被用于治疗蛔虫、肠道蠕虫、寄生虫和胃肠道问题,剂量为连续服用三到四天。此外,墨西哥自然历史学会指出,美洲蒲桃种子粉被用作一种驱虫剂和抗菌剂。另一方面,幽门螺杆菌感染是导致人类胃部疾病、消化性溃疡以及某些类型的胃淋巴瘤和胃癌的一个因素;因此,需要有关于美洲蒲桃种子对胃肠道疾病影响的科学证据。

研究目的

本研究旨在评估美洲蒲桃种子在口服-胃消化过程中的生物可利用性和抗幽门螺杆菌的生物活性,使用体外 INFOGEST 方法对美洲蒲桃种子的食物成分进行研究,并使用计算工具阐明可能的作用机制。

材料和方法

最初,使用超声和浸提分别从美洲蒲桃种子中提取美洲蒲桃籽油和美洲蒲桃种子水提物,并获得美洲蒲桃种子的食物成分。样品通过 INFOGEST 方法进行口服-胃消化,然后通过 HPLC-DAD 和 GC-MS 继续鉴定和定量生物活性化合物。通过最小抑菌浓度测定法、幽门螺杆菌生长抑制动力学测定法和尿素酶抑制动力学测定法,使用 14 株临床分离株和参考株的幽门螺杆菌对其抗幽门螺杆菌活性进行了测定。最后,使用计算工具(AutoDock 4.2.6 软件和 BioVia Discovery v.19.1.0.1.18287)阐明可能的作用机制。

结果

美洲蒲桃种子的亲脂部分检测到油酸、亚油酸和鳄梨呋喃化合物,而酚类部分则显示出儿茶素、芦丁、鞣花酸和绿原酸等物质的存在。在口服-胃消化过程中,由于机械和酸水解作用,酚类化合物发生构象变化,而亲脂性化合物在胃相中增加了 20%。美洲蒲桃种子成分(3.08μg/mL)对临床和参考株幽门螺杆菌均显示出总体外抑制作用,此外,亲脂部分的抑制浓度(2.59μg/mL)较低,与菌株无关。在计算机模拟中发现的机制包括抑制 CagA、BabA 和 MUC5 等靶蛋白,这些靶蛋白是涉及粘附和细菌致病性的毒力因子。

结论

本研究提供了证据表明,美洲蒲桃种子的食物成分具有抗幽门螺杆菌的体外抗菌潜力,这是通过酚类化合物和主要是亲脂性化合物实现的,为支持美洲蒲桃种子的传统用途提供了新的科学证据。

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