Abstract
Trichophyton rubrum (T. rubrum) is one of the most important agents of dermatophyte infection in humans. The aim of this experiment was to evaluate the effect of HaCaT cells on T. rubrum, investigate the responsible mechanism of action, and explore the role of reactive oxygen species (ROS) and nitric oxide (NO) in the inhibition of T. rubrum growth by HaCaT cells. The viability of fungi treated with HaCaT cells alone and with HaCaT cells combined with pretreatment with the NADPH oxidase inhibitor (DPI) or the nitric oxide synthase (NOS) inhibitor L-NMMA was determined by enumerating the colony-forming units. NOS, ROS, and NO levels were quantified using fluorescent probes. The levels of the NOS inhibitor asymmetric dimethylarginine (ADMA) were determined by enzyme-linked immunosorbent assay (ELISA). Micromorphology was observed using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). In addition, fungal keratinase activity was assessed by measuring dye release from keratin azure. In vitro fungal viability, keratinase activity, and ADMA content decreased after HaCaT cell intervention, whereas the levels of ROS, NO, and NOS increased. The micromorphology was abnormal. Fungi pretreated with DPI and L-NMMA exhibited opposite effects. HaCaT cells inhibited the growth and pathogenicity of T. rubrum in vitro. A suggested mechanism is that ROS and NO play an important role in the inhibition of T. rubrum growth by HaCaT cells.
Highlights
Trichophyton rubrum (T. rubrum) is a dermatophyte responsible for the majority of fungal infections worldwide [1] and accounts for as many as 69.5% of dermatophytoses in humans [2]
We aimed to investigate the mechanism stimulated when T. rubrum attacks its host and sought to determine whether reactive oxygen species (ROS) and nitric oxide (NO) play a role in inhibiting the growth of T. rubrum by HaCaT cells, a human keratinocyte cell line
The mechanism that is active in fungi in response to keratinocytes is unclear
Summary
Trichophyton rubrum (T. rubrum) is a dermatophyte responsible for the majority of fungal infections worldwide [1] and accounts for as many as 69.5% of dermatophytoses in humans [2]. Langerhans cells, neutrophils, and macrophages in the stratum corneum appear to be important in the defense against fungi, such as Malassezia spp. and T. rubrum [4]. The exposure of keratinocytes to T. rubrum leads to induced expression of antimicrobial peptides (AMPs), and increased levels of AMPs may help the host control the growth and spread of T. rubrum and most likely other dermatophytes [5]. We hypothesized that keratinocytes may perform a defensive function against T. rubrum.
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