Study On Antifungal Activity Screening And Mechanism Of Traditional Chinese Medicines

Objective To investigate the fungal effect of traditional Chinese medicine(TCM)against deep fungi to a large scale activity screening,looking for the Traditional Chinese Medicines which has the clinical application value against invasive fungal infections.Method The inhibitory effect of Candida albicans,Cryptococcus neoformans and Aspergillus fumigatus the three common invasive fungi was determined by the minimum antimicrobial concentration(MIC)measured by CLSI recommended micro-liquid dilution method with 75% ethanol ultrasonic extraction.The inhibitory effect of Candida albicans hyphae,the antimicrobial spectrum of the traditional Chinese medicine and the effect with fluconazole against FLC-resistant Candida albicans 103,the common or specific material basis of the anti-invasive fungus were studied.The anti invasive fungal infection effect in vivo.was investigated by mortality of immunosuppressed ICR mice model infected gastrointestinal tract with C.albicans SC5314.Results:The antifungal activity of a total of 1577 TCMs,including 210 kinds of antifungal TCMs from the "Zhong hua ben cao" and "Zhong yao da ci dian" recorded,618 herbs "Chinese Pharmacopoeia"(2015 Edition),122 kinds of Uygur Medicine recorded in "Zhong hua ben cao" Uyghur volume and other 779 kinds of traditional Chinese medicine.were screened systematicallyin vitro against Cryptococcus neoformans,Candida albicans,Aspergillus fumigatus three common invasive fungi.The amount of effective medicines against Candida albicans,Cryptococcus neoformans and Aspergillus fumigatus.are 149,453,25,respectively.There are 21 kinds which show high inhibition of thethree deep fungus.On inhibition of the formation of mycelial,we found 53 kinds which can completely inhibit the mycelial growth at the concentration of 4mg/ml,the Paris chinensis Franch.,Dioscorea tokoro Makino,Cynanchum atratum,Dioscorea nipponica Makino,Veratrum nigrum Linn.,Notopterygium incisumcan completely inhibit the hypha formation below 500 ?g/m L.Glochidion puberum,Distylium racemosum,Quercus infectoria Olivier,Chinese olive,Rosa multiflora Thunb.,Terminalia chebula Retz,Cynanchum atratum,Rhodiola rosea L.,Apatite,Acacia senegal Willd,Rhus coriaria L.Mangifera indica L.,Rehmannia piasezkii Maxim.were reported for the first time,and the traditional Chinese medicines have antifungal activity against Candida albicans,MIC ?250?g/m L and deserve to be studied.Through the chemical composition of cluster analysis,we found that theactive substance ofTCMswhich show high anti fungal effect tend to be concentrated in tannins,alkaloids,saponins,volatile oil The antifungal effect of TCMswhich have more tannins is strong.Isolated directly through the traditional active and active components,we found that the active site of the highest antifungal activity mostly in ethyl acetate and n-butanol fractions,and the antifungal activity of tannin part direct separated is also very strong,we could conclude that the common antifungal active sites is tannin.Saponins are represented by Cynanchum atratum,not isolated active sites of tannin,concentrated in the less polar parts.petroleum ether and dichloromethaneIn mechanism of action research,the ultrastructure,apoptosis,active oxygen content determination of the Candida albican was studied by active compounds Geranium 16 ?g/ml,gallic acid 64?g/ml,tannic,acid 4?g/ml and methylene chloride of traditional Chinese medicine.Cynanchum atratum 4?g/ml.The results showed that the ultrastructures of the groups were significantly different,the content of ROS and the apoptosis rate of Geranium was significantly higher than that in the control group.Which ROS was 1.79 times and the apoptosis rate was 27.4%,the other treated groups had no difference than the control group.In vivo animal research,we chosed immunosuppressed ICR mice model infected gastrointestinal tract with C.albicans SC5314.as the main model and dose reference pharmacopoeia,looking for the TCMs which can direct inhibit candida albican or enhance immunity of the body in vivo.Then the results shows some TCMs which show great antifungal effect in vitro,such as Cynanchum atratum,Quercus infectoria Olivier,Polygonum bistorta L.,Red Paeony Root,Rhodiola rosea L.,Distylium racemosum Sieb.et Zucc.,Veratrum nigrumL.,Gynura segetum(Lour.)Merr.show no antifungal activity in vivo,have no ability to improve the survival rate of mice and prolong the survival time.Rhus chinensis Mill(0.1g / kg,0.33 g / kg and 1.09 g / kg),Coptis chinensisFranch.0.58g/kg,Phellodendron chinenseSchneid1.25g/kg can significantly reduce the mortality of mice,prolong the survival time of mice.In tonic TCMs,Astragalus membranaceus(Fisch.)Bunge(2.25g/kg and 5g/kg)can significantly improve the survival time,the other herbs Common Yam Rhizome,Rehmannia glutinosa(Gaert.)Libosch.ex Fisch.et Mey.,Schisandra chinensis,Codonopsis pilosula(Franch.)Nannf.,Atractylodes macrocephala Koidzhad no differences with the control group in the corresponding dose of pharmacopoeia.Traditional Chinese medicine active natural monomer tannin(tannic acid 1mg/kg,gallic acid10mg/kg Geraniin 1mg/kg)could significantly prolong the survival time of mice,and have significant difference with the control group.Conclusion For the first time we have a large-scale antifungal activity screening for the 1577 traditional Chinese medicines with a unified standard in vitro,and firstly found the TCM,such as Turkish gall,Chinese olive,Terminalia chebula Retz,Maobian Zi,Acacia senegal Willd,Rhus coriaria l.fruit with antifungal activity,which have found that tannin is their common material basis.The in vivo antifungal activity of some Chinese medicine is verified,and the antifungal activity of the synergistic clinical drug is studied,which provides some reference for clinical medication guidance.Traditional Chinese medicine is worth exploring...