Cordeiro Rossana de Aguiar, Serpa Rosana, Mendes Patrícia Bruna Leite, Evangelista Antonio José de Jesus, Andrade Ana Raquel Colares, Franco Jônatas da Silva, Pereira Vandbergue Dos Santos, Alencar Lucas Pereira de, Oliveira Jonathas Sales de, Camargo Zoilo Pires de, Lima Neto Reginaldo Gonçalves de, Castelo-Branco Débora de Souza Collares Maia, Brilhante Raimunda Samia Nogueira, Rocha Marcos Fabio Gadelha, Sidrim José Júlio Costa
a Medical Mycology Specialized Center , Federal University of Ceará , Fortaleza , Brazil.
b Department of Microbiology, Immunology and Parasitology , Federal University of São Paulo , São Paulo , Brazil.
Biofouling. 2017 Sep;33(8):640-650. doi: 10.1080/08927014.2017.1350947.
This study evaluated the effect of the protease inhibitor ritonavir (RIT) on Trichosporon asahii and Trichosporon inkin. Susceptibility to RIT was assessed by the broth microdilution assay and the effect of RIT on protease activity was evaluated using azoalbumin as substrate. RIT was tested for its anti-biofilm properties and RIT-treated biofilms were assessed regarding protease activity, ultrastructure and matrix composition. In addition, antifungal susceptibility, surface hydrophobicity and biofilm formation were evaluated after pre-incubation of planktonic cells with RIT for 15 days. RIT (200 μg ml) inhibited Trichosporon growth. RIT (100 μg ml) also reduced protease activity of planktonic and biofilm cells, decreased cell adhesion and biofilm formation, and altered the structure of the biofilm and the protein composition of the biofilm matrix. Pre-incubation with RIT (100 μg ml) increased the susceptibility to amphotericin B, and reduced surface hydrophobicity and cell adhesion. These results highlight the importance of proteases as promising therapeutic targets and reinforce the antifungal potential of protease inhibitors.
本研究评估了蛋白酶抑制剂利托那韦（RIT）对阿萨希毛孢子菌和因克毛孢子菌的作用。通过肉汤微量稀释法评估对RIT的敏感性，并以偶氮白蛋白为底物评估RIT对蛋白酶活性的影响。测试了RIT的抗生物膜特性，并评估了经RIT处理的生物膜的蛋白酶活性、超微结构和基质组成。此外，在浮游细胞与RIT预孵育15天后，评估了抗真菌敏感性、表面疏水性和生物膜形成情况。RIT（200μg/ml）抑制毛孢子菌生长。RIT（100μg/ml）还降低了浮游细胞和生物膜细胞的蛋白酶活性，减少了细胞黏附和生物膜形成，并改变了生物膜的结构和生物膜基质的蛋白质组成。用RIT（100μg/ml）预孵育可增加对两性霉素B的敏感性，并降低表面疏水性和细胞黏附。这些结果突出了蛋白酶作为有前景的治疗靶点的重要性，并加强了蛋白酶抑制剂的抗真菌潜力。
