Cordeiro Rossana de Aguiar, Evangelista Antonio José de Jesus, Serpa Rosana, Marques Francisca Jakelyne de Farias, Melo Charlline Vládia Silva de, Oliveira Jonathas Sales de, Franco Jônatas da Silva, Alencar Lucas Pereira de, Bandeira Tereza de Jesus Pinheiro Gomes, Brilhante Raimunda Sâmia Nogueira, Sidrim José Júlio Costa, Rocha Marcos Fébio Gadelha
Specialized Medical Mycology Center,Federal University of Ceará,Fortaleza, CE, Brazil.
Postgraduate Program in Medical Sciences,Federal University of Ceará, Fortaleza, CE,Brazil.
Microbiology (Reading). 2016 Feb;162(2):309-317. doi: 10.1099/mic.0.000222. Epub 2015 Dec 8.
Heat-shock proteins (Hsps) are chaperones required for the maintenance of cellular homeostasis in different fungal pathogens, playing an important role in the infectious process. This study investigated the effect of pharmacological inhibition of Hsp90 by radicicol on the Cryptococcus neoformans/Cryptococcus gattii species complex--agents of the most common life-threatening fungal infection amongst immunocompromised patients. The influence of Hsp90 inhibition was investigated regarding in vitro susceptibility to antifungal agents of planktonic and sessile cells, ergosterol concentration, cell membrane integrity, growth at 37 °C, production of virulence factors in vitro, and experimental infection in Caenorhabditis elegans. Hsp90 inhibition inhibited the in vitro growth of planktonic cells of Cryptococcus spp. at concentrations ranging from 0.5 to 2 μg ml(-1) and increased the in vitro inhibitory effect of azoles, especially fluconazole (FLC) (P < 0.05). Inhibition of Hsp90 also increased the antifungal activity of azoles against biofilm formation and mature biofilms of Cryptococcus spp., notably for Cryptococcus gattii. Furthermore, Hsp90 inhibition compromised the permeability of the cell membrane, and reduced planktonic growth at 37 °C and the capsular size of Cryptococcus spp. In addition, Hsp90 inhibition enhanced the antifungal activity of FLC during experimental infection using Caenorhabditis elegans. Therefore, our results indicate that Hsp90 inhibition can be an important strategy in the development of new antifungal drugs.
热休克蛋白(Hsps)是不同真菌病原体维持细胞稳态所需的伴侣蛋白,在感染过程中发挥重要作用。本研究调查了萝卜硫素对Hsp90的药理抑制作用对新生隐球菌/格特隐球菌复合种的影响,该复合种是免疫功能低下患者中最常见的危及生命的真菌感染的病原体。研究了Hsp90抑制对浮游细胞和固着细胞对抗真菌药物的体外敏感性、麦角固醇浓度、细胞膜完整性、37℃下的生长、体外毒力因子产生以及秀丽隐杆线虫实验感染的影响。Hsp90抑制在0.5至2μg ml(-1)的浓度范围内抑制了隐球菌属浮游细胞的体外生长,并增强了唑类药物,尤其是氟康唑(FLC)的体外抑制作用(P < 0.05)。Hsp90的抑制还增强了唑类药物对隐球菌属生物膜形成和成熟生物膜的抗真菌活性,对格特隐球菌尤为明显。此外,Hsp90抑制损害了细胞膜的通透性,降低了37℃下的浮游生长以及隐球菌属的荚膜大小。此外,在使用秀丽隐杆线虫的实验感染过程中,Hsp90抑制增强了FLC的抗真菌活性。因此,我们的结果表明,Hsp90抑制可能是开发新型抗真菌药物的重要策略。