Study On Mechanism Of Magnolol Against Cryptococcus Neoformans

Objective: In order to sum up experience and improve the level of diagnosis and treatment of pulmonary cryptococcosis,The clinical data of pulmonary cryptococcosis patients was analyzed.Method: Patients definitely diagnosed with pulmonary cryptococcosis in the Frist Affiliated Hospital of Dalian Medical University between October 2003 and July 2021 were selected.The clinical data of all patients with pulmonary cryptococcosis was reviewed.The paper studies the general information、predisposing factors、clinical manifestations、radiological findings、diagnostic and treatments methods.SPSS 22 statistical software was used for analysis.Result: 50 patient of Pulmonary Cryptococcosis were enrolled in the research,males : females is 1:1.The age ranged from 27 to 82 years old.The mean(±SD)age of them was 53.56±11.99 years,Age group of patients,ten years into an age group,the high incidence of ages were 40 to 60 years old.2 cases(4%)had history of exposure to birds.18 cases(36%)accompanied by one or more basic disease,among the basic disease,hypertension were seen more.among the clinical symptoms cough were seen more.Pulmonary opacity and pulmonary mass were most common in the initial diagnosis.among the Imaging performance,nodular and nodular shadows were seen more.Distribution characteristics of lesions: The lower lobe is greater than the upper lobe 、the single lobe is greater than double leaf、the right lung is greater than the left lung.among the,the most common concomitant signs of the lesions is spicule sign.7 patients had pulmonary ventilation dysfunction。In the laboratory examination,there are no specific indicators.Pathological examination mainly showed the appearance of granuloma.47 cases(94%)were diagnosed by biopsy.2 cases(4%)were diagnosed by serological examination.Cryptococcal capsular antigen tests were positive and accompanied by pulmonary lesions.1 cases(2%)was diagnosed by the discovery on a Cryptococcus smear.43 cases(86%)were treated with surgical excision.6 cases(12%)were treated with antifungal agents.1 case(2%)was transferred to another hospital for suspected pulmonary tuberculosis.Conclusion: Pulmonary cryptococcosis is more likely to occur in middle-aged and elderly people.Immunocompetent or immunocompromised persons can both have pulmonary cryptococcosis.Among the clinical symptoms cough was seen more.among the Imaging performance Pulmonary nodule was seen more.The treatment is more depends surgical excision.Cryptococcus neoformans is a conditionally pathogenic fungus containing capsule that can cause severe fungal infectious disease.Due to the overuse of antifungal drugs,the problem of drug resistance is becoming more and more serious,and the search for new antifungal drugs is imminent.Chinese herbal medicine has become a hot spot in recent years due to its wide range of sources and low side effects.Objective: In this project,Cryptococcus neoformans BNCC225501 and clinically isolated strains were used as research objects to explore the antifungal activity of magnolol on the test strains,and the possible molecular mechanism of magnolol against Cryptococcus neoformans BNCC225501 was explored by transcriptomics and metabolomics.The results provide ideas and directions for researchers to develop new antifungal drugs,and provide a theoretical basis for the development of new antifungal drugs based on magnolol.Method: 1.The inhibitory effect of common antifungal drugs on Cryptococcus neoformans was determined by ATB FUNGUS 3 method.2.The inhibitory effect of magnolol on Cryptococcus neoformans was determined by broth dilution method.3.The checkerboard method was used to detect whether the combination of magnolol and the existing drug fluconazole for the treatment of Cryptococcus neoformans had a synergistic effect.4.Cryptococcus neoformans were cultured in RPMI 1640 medium containing different concentrations of magnolol and fluconazole drugs,and the concentration was adjusted to 2×103CFU/m L,and the control group was the DMSO group with drug solvent.Every 12 h,the quantitative bacterial solution was coated with plates and the number of surviving colonies was observed.5.Cryptococcus neoformans were cultured on DMEM,Niger Seed and Urea base culture-medium with magnolol added in these media to detect the effect of magnolol on Cryptococcus neoformans’ s capsule,melanin and urease.6.To study the mechanism of magnolol exerting antifungal activity against Cryptococcus neoformans,the gene transcriptome and metabonomics of Cryptococcus neoformans in magnolol administration group、fluconazole administration group、magnolol fluconazole combined administration group and blank group were detected.Bioinformatics analysis was used to research the change of transcription and metabolism levels.Result: 1.In this study,Cryptococcus neoformans BNCC225501 and clinical isolates were used as the research object to explore the antifungal activity of magnolol on the tested strains.It was preliminatively determined that magnolol had antibacterial activity on Cryptococcus neoformans in culture,affecting the growth of cryptococcus neoformans,and it was clear that magnolol could inhibit the capsule size of cryptococcus neoformans and the ability to synthesize urease.2.Taking Cryptococcus neoformans BNCC225501 as the research object,transcriptomic exploration found that compared with the control group,the magnolol treated Cryptococcus neoformans mainly affected biological functions or pathways such as transmembrane transport,redox process,inherent components of membranes,membrane overall components,drug binding,hydrolase activity,oxidoreductase activity,pyrophosphatase activity,DNA replication,meiosis-yeast,fatty acid biosynthesis.Compared with the blank group,Cryptococcus neoformans treated with the combination of magnolol fluconazole mainly affected biological functions or pathways such as transmembrane transport,redox process,intrinsic components of membrane,membrane wholeness,transport activity,transmembrane transport activity,oxidoreductase activity,nucleoside triphosphate activity,hydrolase activity,DNA replication,glycine,serine and threonine metabolism,NOD-like receptor signaling pathway.3.Taking Cryptococcus neoformans BNCC225501 as the research object,metabolomics exploration was used to find that the metabolic pathways of Cryptococcus neoformans treated with magnolol mainly affected histidine metabolism and arginine biosynthesis.In the comparison between the combination group of magnolol fluconazole and the blank group,the main metabolic pathways affected were histidine metabolism,sphingomyelin metabolism,arginine biosynthesis.Conclusions: 1.Magnolol has bacteriostatic and bactericidal ability for Cryptococcus neoformans,and can inhibit the capsule size and urease production capacity of Cryptococcus neoformans.2.The mechanism of action of magnolol antibacterial may be by inhibiting the bacterial detoxification response by affecting the content of glutathione and glutathione transferase.