Screening Of Antifungal And Antitumor Active Components In Trichoderma Serrata

Dryopteris fragrans(L.)schott is a deciduous perennial herb of the Dryopteris genus,which grows mainly in the talc slope of the alpine region,the scree of the forest and lava cracks around volcanoes.According to the history records in Wu-da-lian-chi,Dryopteris fragrans has been used to treat many diseases by minority hundred years ago,especially for skin diseases.Trichophyton,Microsporon and Epidermophyton are three main pathogenic fungi giving rise to fungal skin diseases.Microsporum ferrugineum and Microsporum gypseum belong to Microsporon,which is a zoonotic pathogen of skin infection.Trichophyton rubrum and Trichophyton tonsurans are easy to cause kerion,tineacorporis,tineacapitis,tineabarbae and strong inflammatory reaction and multiple nodules.Epidermophyton floccosum can easily infect healthy immune system of host cuticle tissues,such as nails,hairs and skins etc.Malignancy is the main disease threatening human health and life nowadays.There are various of chemical synthetic drugs curing for cancer,howere with the drugs more available,drug resistance occurs clinically.Accordingly,the key point of this study is exploreing novel anticancer drugs with low toxicity and high efficiency from nature products.In this paper,the water extract was obtained from Dryopterisfragrans,and was subjected to DM130 macroporous resin column eluted by 20%ethanol,40%ethanol,65%ethanol and 95%ethanol,respectively.The corresponding eluted fractions of the extract were tested with antifungal and antitumour activity.The results showed that 40%ethanol fraction had the best antifungal and anti-tumor effect.The MIC of Trichophyton rubrum,Trichophyton tonsurans,Epidermophyton floccosum and Microsporum gypseum was 12.5-25 mg/m L,3.13-6.25 mg/m L,6.25-12.5 mg/m L,12.5-25 mg/m L,respectively.But these four eludted fractions had no obvious inhibition effect on Microsporum ferrugineum.The MFC of Trichophyton rubrum,Trichophyton tonsurans,Epidermophyton floccosum and Microsporum gypseum was 25-50 mg/m L,12.5-25mg/m L,12.5-25 mg/m L,25-50 mg/m L,respectively.Antitumor activity illustrated that four fractions possessed varying degrees of inhibition effects on human lung cancer cell A549,human gastric cancer cell SGC-7901 and human breast cancer cells MCF-7,especially 40%ethanol fraction,which had remarkable inhibitory activity.Cell inhibition rates were 66.5%,63.6%and 70.9%,respectively.Therefore,it was determined that 40%ethanol eluted fraction aimed at antifungal and antitumor.Normal phase silica gel column chromatography,reversed-phase(MCI,ODS)column chromatography,Sephadex LH-20 column chromatography,thin-layer chromatography and high-performance liquid chromatography technique were used to treat 40%ethanol fraction from which sixteen monomeric compounds were purified.and finally their structure were determined by multi-spectrum and NMR and combined with literatures reported before.Their structural types include five phenylpropanoid compounds,three flavonoids,two coumarins,two aromatic compounds,two lignanoids,one iridoid glycosides and one sterol.The result of antifungal activity test showed that dryopteris A,dryopteris K,caffeic acid,quercetin,p-hydroxybenzoic acid,aesculetin,coumaric acid,?-sitosterol had different degrees of inhibiton efffect on five test fungi.As was shown in the results,dryopteris K only had a certain inhibitory effect on Microsporum ferrugineum and dryopteris A only had a certain extent inhibitory effect on Trichophyton rubrum.The present study provides a new medical resource for further study on treatment of skin diseases caused by above pathogenic fungi.The results of antitumor activity displayed that Dryopteris A,coumaric acid,caffeic acid had the most significant inhibition compared with negative control in A549 cells with the inhibitory rate of 79.3%?76.9%?63.5%,respectively.IC₅₀ values were 37.98±1.58?mol/L,25.79±1.41?mol/L,19.86±1.30?mol/L;14.92±1.17?mol/L,10.41±1.02?mol/L,8.55±0.93?mol/L;13.65±1.14?mol/L,9.77±0.99?mol/L,8.18±0.91?mol/L at 24h,48h and 72h.Dryopteris K,quercetin and quercetin-3-O-?-D-glucoside and possessed the most significant activities compared with negative control in MCF-7 cells with inhibitory rate of 78.6%?80.1%?75.7%,respectively.IC₅₀ values were 29.32±1.47?mol/L,20.57±1.31?mol/L,13.81±1.14?mol/L;17.06±1.23?mol/L,11.42±1.06?mol/L,9.64±0.98?mol/L;27.71±1.44?mol/L,23.34±1.37?mol/L,16.80±1.23?mol/L at 24h,48h and 72h.Quercetin,?-sitosterol and caffeic acid had the most significant activities compared with negative control in SGC-cells,and the inhibitory rate was 69.8%?65.8%?72.3%,respectively.IC₅₀ values were 32.60±1.51?mol/L,19.37±1.29?mol/L,13.67±1.14?mol/L;30.04±1.48?mol/L,20.46±1.31?mol/L,15.14±1.18?mol/L;15.54±1.19?mol/L,8.96±0.99?mol/L,7.56±0.88?mol/L at 24h,48h and 72h.All of these activities were related to the concentration and time of the compounds.