| T.rubrum is a kind of Dermatophytes,which can infect human skin,nails and hair.In recent years,the incidence of clinical fungal infections have increased year by year due to the widespread use of broad-spectrum antibiotics,corticosteroids,and immunosuppressants.Allylamines and azole are commonly used to treat fungal infections,but there are different degrees of side effects,and due to the high recurrence of fungal infection,treatment effect has reduced.At present,the new therapeutic drugs are almost at a standstill,while fungal resistance is becoming more and more serious.Therefore,finding new antifungal agents is of great significance.Linalool derived from natural products,is widely used in cosmetics and food industry,with many biological activities,such as antianxiety,antiinflammation,anticonvulsion,antibiosis.We put linalool as a research object to study its antifungal activity and mechanism on T.rubrum,and providing the basis for searching a new antifungal drugs.Objective:The main research of this subject is the antifungal activity and mechanism of linalool on T.rubrum,on the basis of guinea-pig T.rubrum model to evaluate the therapeutic effect of linalool and contribute to intensive study the antifungal mechanism of linalool and the development of antifungal agents.Methods:(1)Antifungal activity of linalool in vitro.Filter paper dispersion method is used to test the antifungal effect of linalool in T.rubrum,C.albicans,M.gypseum.The modified CLSI M38-A2 method is used to analyze the minimum inhibitory concentration of linalool on T.rubrum.(2)Study on the antifungal mechanism of linalool on T.rubrum.After the action of linalool observing mycelial morphology changes and conidia germination of T.rubrum by microscope.Comparing the radial growth diameter of mycelium by cross method;According to the conductivity changes and PI staining results to analysis the changes of cell permeability and research the influence on synthesis of ergosterol,then analysising the micro structure changes of T.rubrum by TEM.(3)The orthogonal experimental design of linalool gel.Carbopol,propylene glycol and glycerol as the test factors of orthogonal design to optimization linalool gel.(4)The construction of the guinea pig model of T.rubrum and the curative effect of linalool.Firstly,T.rubrum infect the back skin of guinea pigs then evaluating the therapeutic effect of linalool gel.Results:(1)Filter paper dispersion method results showed that inhibitory effect of linalool in T.rubrum,C.albicans,M.gypseum,the inhibition zone diameter were 15.6 mm,12 mm,18.3 mm;T.rubrum was high sensitive to linalool,C.albicans was extremely sensitive and M.gypseum was Middle sensitive to it.According to modified CLSI M38-A2 that the MIC of linalool is 320 ?g/m L.(2)After the treatment of linalool,the mycelium of T.rubrum shrunk and rough,top and local necrosis,cytoplasm autolysis;conidia germination and mycelial radial growth were inhibited which increased with the concentration of liquid medicine.The permeability changed and synthesis of ergosterol reduced;microscopic structure has been changed which manifested in cell wall degradation,surface roughness anduneven thickness,cell intima dilapidation,unclear contour,the damage of some organelles expressed as membrane fragments and cytoplasm leakage.(3)Orthogonal experiment results for the optimal formula of linalool gel is carbomer 0.8 g,glycerol 5 g,propylene glycol 0.4 g,linalool 0.032 g,triethaolamine 1.08 g,replenishing water to 100 g.(4)The guinea pig model of T.rubrum was successfully constructed.After the treatment of linalool,dander significantly reduced,continuous medication for 10 d,dander disappeared completely and returned to normal.Compared with the model group found the structure of the epidermis and the dermis was evenly distributed,the degree of tissue destruction was reduced,the epidermis keratinization degree was normal,thickness of the spine layer was uniform,without dilatation and fusion in hair follicle and the skin was basically recovered. |
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