Study On The Mechanism And Application Of Active Ingredients From Platycladus Orientalis Leaves Against Trichophyton Rubrum Membran

[Objective] To isolate and identify the main active ingredients of Cacumen Platycladi against Trichophyton rubrum(T.rubrum)and to clarify its antifungal membrane mechanism of action,to prepare its nanoemulsion gel to verify the clinical efficacy,and to provide theoretical support for the development of new antifungal drugs and clinical use.[Methods] The main active ingredients and membrane mechanism of action of T.rubrum were screened and verified by activity tracking method,micro-broth dilution method,GC-MS,spore germination test,electron microscopy,flow cytometry,qualitative analysis of ergosterol and q RT-PCR;α-pinene nano-emulsion gel was prepared and characterized and tested for skin irritation;T.rubrum-guinea pig model to test the effect of α-pinene nanoemulsion gel in the treatment of T.rubrum infection.[Results] α-pinene from the petroleum ether site of Cacumen Platycladi was the main anti-T.rubrum active ingredient with MIC of 32 μg/m L,which could inhibit spore germination significantly(p< 0.01).α-pinene could cause a significant decrease in ergosterol content of T.rubrum cell membrane(p< 0.05),resulting in significant damage and a highly significant increase in permeability(p< 0.01),in sterol The mass spectra represented the presence of marker sterols with inhibited ERG3 content or activity,and ERG3 expression was highly significantly reduced(p< 0.01);the p H of each α-pinene nanogel prepared was neutral(6.8±0.4),the particle size distribution was between 17.81 and 22.56 nm,and the zeta potential was between-8.26 and-6.98 mv,which met the nanoformulation The above indexes did not change significantly under extreme conditions(p> 0.05),and there was no skin irritation.The clearance rate of α-pinene nanoemulsion gel preparation for T.rubrum was 74.39%,and the clinical efficiency was 67.34%,which can effectively remove T.rubrum spores harbored in hair follicles and hair shaft medulla,and has good therapeutic effect on them.[Conclusion]α-pinene is the main active ingredient of T.rubrum in Cacumen Platycladi,which interferes with the synthesis of ergosterol of T.rubrum with ERG3 as the action target and thus disrupts the cell membrane to exert antifungal effect,and its nano-emulsion gel is a highly stable,safe and obvious nano-formulation for curing T.rubrum infection.