Effects of Aspirin as an Anti-inflammatory Drug on Azole-resistant Candida glabrata In Vitro

Candida glabrata previously considred as a nonpa-thogenic commensal microorganism of human mucosal tissues, is now mentioned as the second or third cause of nosocomial candidiasis and have significantly increased recently due to immuno-suppressive therapies (1). Among the Candida species, C. glabrata is inherently resistant to azole anti-fungal agents. The mutations occurred in several ergosterol biosynthesis genes including ERG1, ERG3, ERG6, ERG7, ERG9 and ERG11 confer the resistance to azoles (2). The nonessen-tial gene ERG6, undergoes mutations can develop multiple phenotypes, including, decreased er-gosterol content, increased resistance to polyenes and increased cycloheximide sensitivity (3). The non-steroidal anti-inflammatory drugs (NSAID) especially ibuprofen diclofenac and aspirin have been indicated to have inhibitory effect on bio-film formation and decrease of drug resistance in C. albicans (4). The aim of this investigation was to assess the effects of aspirin as an anti-inflammatory drug on the C. glabrata species in vitro. A resistance isolate of C. glabrata obtained from patients with vulvo vaginal candidiasis was selected for further analysis in this study. The antibiotic susceptibility test of C. glabrata isolates for fluconazole (Pfizer Central Research, Sandwich, United Kingdom) according to the CLSI M27 A3 standard protocol. The MICs endpoints of fluconazole as the level, induced a prominent reduction of growth (50% inhibition), compared to drug-free growth control. Based on this method, strains considered as susceptible when the MIC was ≤ 8 μg/ml, considered as susceptible dose dependent when MIC was =16–32 μg/ml, and considered as resistant when MIC was ≥64 μg/ml. One fluconazole-resistant isolate has been chosen for further investigation. The effect of aspirin (Bayern, Germany) on the growth as well as gene regulation of the mentioned fluconazole-resistant isolate was performed. Briefly, according to a serial dilution process, an initial dilution of aspirin (250 mg/ml) was used for serial dilutions (125, 62.5, 31.25, 15.62, 7.81 mg/ml). Inoculum suspensions were prepared from 24 h of Candida cultures ranged from 2.5×10 3 0.5×10 3 to CFU/ml by spectrophotometry. The plates were then incubated at 35 °C for 48 h. The plates were incubated in 35 °C for 48 h then visual readings were performed. For the confirmation of growth of isolates, 10 µl of each plate were inoculated to the sabouraud's dextrose agar plates.