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从辣木种子中提取的纯化几丁质结合蛋白 Mo-CBP 是一种有效的抗皮肤真菌蛋白:体外作用机制、体内抗感染效果以及作为皮肤感染水凝胶的临床应用。

Mo-CBP, a purified chitin-binding protein from Moringa oleifera seeds, is a potent antidermatophytic protein: In vitro mechanisms of action, in vivo effect against infection, and clinical application as a hydrogel for skin infection.

机构信息

Department of Biochemistry and Molecular Biology, Federal University of Ceará, Fortaleza, Ceará, Brazil.

Department of Biochemistry and Molecular Biology, Federal University of Ceará, Fortaleza, Ceará, Brazil.

出版信息

Int J Biol Macromol. 2020 Apr 15;149:432-442. doi: 10.1016/j.ijbiomac.2020.01.257. Epub 2020 Jan 28.

Abstract

Dermatophytes belonging to Trichophyton ssp. are important anthropophilic and zoophilic pathogens, which developed resistance to griseofulvin, the common antifungal drug used to treat dermatophytosis. In this context, Moringa oleifera seed proteins have been described as antifungal agents with potential applications. Thus, this work aimed to evaluate the antidermatophytic in vitro, focusing on mechanisms, and in vivo potential of Mo-CBP, purified from M. oleifera seeds. Mo-CBPwas purified after protein extraction with 50 mM Tris-HCl buffer, pH 8.0, and chromatography on chitin and CM Sepharose™ columns and antidermatophytic potential of Mo-CBP evaluated in vitro and in vivo. In vitro, Mo-CBP reduced in 50% the germination of microconidia of Trichophyton mentagrophytes at 45 μM; but did not show inhibition of mycelial growth. Mo-CBP (45 μM) presents the inhibitory activity even when incubated with N-acetyl-d-glucosamine (NAG). Analysis of the mechanisms of Mo-CBP revealed an increase in membrane permeability, ROS overproduction and damage to cell wall leading to microconidia death. Furthermore, using in vivo models, Mo-CBP (5, 10 and 20 mg g) reduced the severity and time of dermatophytosis. Altogether, these findings indicate that Mo-CBP has great potential for the development of novel antifungal drugs for the clinical treatment of dermatophytosis.

摘要

属于毛癣菌属的皮肤癣菌是重要的亲动物性和亲动物性病原体,它们对灰黄霉素产生了耐药性,灰黄霉素是一种常用的治疗皮肤癣菌病的抗真菌药物。在这种情况下,辣木籽蛋白已被描述为具有潜在应用的抗真菌剂。因此,本工作旨在评估辣木籽蛋白(Mo-CBP)的体外抗皮肤癣菌活性,重点关注其作用机制和体内潜力。Mo-CBP 是从辣木籽中提取蛋白质后,用 50mM Tris-HCl 缓冲液(pH8.0)进行蛋白提取,然后用几丁质和 CM Sepharose™柱进行层析纯化而得到的。体外,Mo-CBP 在 45μM 时将 Trichophyton mentagrophytes 的微孢子萌发率降低了 50%;但对菌丝生长没有抑制作用。Mo-CBP(45μM)即使在与 N-乙酰-d-葡萄糖胺(NAG)孵育时也具有抑制活性。Mo-CBP 作用机制分析表明,它会增加细胞膜通透性、产生过多的 ROS 并破坏细胞壁,导致微孢子死亡。此外,使用体内模型,Mo-CBP(5、10 和 20mg·g)降低了皮肤癣菌病的严重程度和发病时间。综上所述,这些发现表明 Mo-CBP 具有开发新型抗真菌药物用于临床治疗皮肤癣菌病的巨大潜力。

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