The Study Of The Effect Of 420nm Intense Pulsed Light On Trichophyton Rubrum In Vitro

[Background]Light therapy has certain inhibitory effects on microorganisms directly and generates abundant intracellular ROS for oxidative damage.It is speculated that Intense pulsed light(IPL)has a similar effect.In eukaryotes,nicotinamide adenine dinucleotide phosphate oxidase(NADPH oxidase,Nox)can induce the generation of ROS.NADPH oxidase in fungus also induces ROS increase,leading to oxidative stress injury.The study found that the LED could inhibit T.rubrum,causing oxidative stress and nitrosative stress.Therefore,we will investigate whether IPL inhibites T.rubrum by nitrosative stress.In this study,the use of IPL420nm in vitro intervention T.rubrum and explore its effect.[Objective]We detect the expression of ROS and NO,NADPH oxidase and nitric oxide synthase(NOS)and keratinase activity in T.rubrum to investigate the inhibitory effect of intense pulsed light on T.rubrum for providing a theoretical basis of the IPL420nm treatment of T.rubrum and other superficial fungal infectious diseases.[Methods](1)Cultured Trichophyton rubrum(clinical strain and standard strain ATCC4438),divided into 2 groups:T.rubrum,T.rubrum+IPL,respectively,do energy-dependent and time-dependent testing,select the optimal energy.(2)According to the above,the experiment was divided into three groups:T.rubrum,T.rubrum + IPL,T.rubrum + DPI(NADPH oxidase inhibitor)+ IPL.Detection of fungal activity,detection of ROS and MDA,and the levels of SOD and GSH-PX were measured.The expression of NADPH oxidase was detected.Fungal morphology was observed by scanning electron microscopy.The changes of fungal keratin were detected.(3)According to the above,the experiment was divided into three groups:T.rubrum,T.rubrum + IPL,T.rubrum + L-NMMA(NOS inhibitor)+ IPL.MTT detection of fungal activity,and the ROS were measured.The expression of NOS was detected.Fungal morphology was observed by scanning electron microscopy.The changes of fungal keratin were detected.[Results](1)T.rubrum showed dose-dependent and time-dependent growth inhibition,after treatment with IPL.(2)After treatment with IPL,the levels of ROS in the fungus increased,the content of MDA increased,the activity of SOD,GSH-PX and keratinase decreased,the expression of Nox gene was up-regulated,and electron microscopy showed that the fungi of IPL group were seriously damaged.DPI can inhibit IPL processing.(3)After treatment with IPL,the level of NO in the fungus was increased and the expression of NOS was up-regulated.L-NMMA can inhibit IPL processing.[Conclusions](1)IPL420nm can induce ROS and NO excessive production,resulting in oxidation and nitriding damage,and inhibit T.rubrum growth.(2)IPL420nm can inhibit T.rubrum growth by NADPH oxidase and nitric oxide synthase;(3)The study can provide a theoretical basis that intense pulsed light may be a safe and new method to treat superficial fungal infectious diseases.