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德维尔德对临床分离株具有良好的抗真菌活性:对成年白化大鼠肾皮质的体外和体内影响

Promising Antifungal Activity of de Wild against Clinical Isolates: In Vitro and In Vivo Effects on Renal Cortex of Adult Albino Rats.

作者信息

Negm Walaa A, El-Aasr Mona, Attia Ghada, Alqahtani Moneerah J, Yassien Rania Ibrahim, Abo Kamer Amal, Elekhnawy Engy

机构信息

Department of Pharmacognosy, Faculty of Pharmacy, Tanta University, Tanta 31527, Egypt.

Department of Pharmacognosy, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia.

出版信息

J Fungi (Basel). 2022 Apr 21;8(5):426. doi: 10.3390/jof8050426.

Abstract

can cause various infections, especially in immunocompromised patients. Its ability to develop resistance to the current antifungal drugs as well as its multiple virulence factors have rendered the problem even more complicated. Thus, in the present investigation, we elucidated an in vitro and in vivo antifungal activity of methanol extract (ELME) against clinical isolates for the first time. A phytochemical identification of 64 compounds was conducted in ELME using LC-MS/MS. Interestingly, ELME exhibited antifungal activity with MIC values that ranged from 32-256 µg/mL. Furthermore, we investigated the antibiofilm activity of ELME against the biofilms formed by isolates. ELME displayed antibiofilm activity using a crystal violet assay as it decreased the percentages of cells, moderately and strongly forming biofilms from 62.5% to 25%. Moreover, the antibiofilm impact of ELME was elucidated using SEM and fluorescent microscope. A significant reduction in the biofilm formation by isolates was observed. In addition, we observed that ELME resulted in the downregulation of the biofilm-related tested genes (, , , and ) in 37.5% of the isolates using qRT-PCR. Besides, the in vivo antifungal activity of ELME on the kidney tissues of rats infected with was investigated using histological and immunohistochemical studies. ELME was found to protect against induced renal damage, decrease desmin and inducible nitric oxide synthase, increase alkaline phosphatase, and increase infected rats' survival rate. Additionally, the cytotoxicity of ELME was elucidated on Human Skin Fibroblast normal cells using MTT assay. ELME had an IC of 31.26 µg/mL. Thus, we can conclude that ELME might be a promising future source for antifungal compounds.

摘要

可引起各种感染,尤其是在免疫功能低下的患者中。其对当前抗真菌药物产生耐药性的能力以及多种毒力因子使问题更加复杂。因此,在本研究中,我们首次阐明了甲醇提取物(ELME)对临床分离株的体外和体内抗真菌活性。使用LC-MS/MS对ELME中的64种化合物进行了植物化学鉴定。有趣的是,ELME表现出抗真菌活性,MIC值范围为32 - 256 µg/mL。此外,我们研究了ELME对分离株形成的生物膜的抗生物膜活性。ELME使用结晶紫测定法显示出抗生物膜活性,因为它将中度和强烈形成生物膜的细胞百分比从62.5%降低到了25%。此外,使用扫描电子显微镜(SEM)和荧光显微镜阐明了ELME的抗生物膜作用。观察到分离株形成的生物膜有显著减少。此外,我们使用qRT-PCR观察到ELME在37.5%的分离株中导致生物膜相关测试基因(, , ,和 )的下调。此外,使用组织学和免疫组织化学研究调查了ELME对感染 的大鼠肾脏组织的体内抗真菌活性。发现ELME可预防 诱导的肾损伤,降低结蛋白和诱导型一氧化氮合酶水平,增加碱性磷酸酶水平,并提高感染大鼠的存活率。此外,使用MTT测定法阐明了ELME对人皮肤成纤维细胞正常细胞的细胞毒性。ELME的IC为31.26 µg/mL。因此,我们可以得出结论,ELME可能是未来抗真菌化合物的一个有前景的来源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b0c/9144060/467c4ad818f2/jof-08-00426-g002.jpg

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