| Dryopteris fragrans (L.) schot is a deciduous perennial herb of the Dryopterisgenus, which grows mainly in the talc slope of the alpine region, the scree of the forestand lava cracks around volcanoes, mainly distributed in Northeast China, North China,Russia, Japan, Korea, Europe and America. According to the local history records inWu-Da-Lian-Chi, decoction of D. Fragrans has been used to treat many diseases byDaur people for hundreds of years, especially for all kinds of skin diseases. Previousresearch has reported its pharmacological actions, that is it can inhibit different kinds oftumors.Main pathogenic bacteria causing Fungal diseases are Trichophyton, Microsporon,Epidermophyton. Trichophyton tonsurans is one of Trichophyton genus, which cancause children tinea nigra, tinea unguium, tinea of feet and hands and tinea corporis, etc.It is one of velvet pathogen for human. Microsporum gypseum is one of Microsporumgenusas, which can cause animal tinea favosa and human tinea corporis, tinea kerion,and tinea alba, etc.Malignant tumor is one of the most hazardous and severest diseases. Since clinicaltreatment for malignant tumor at present largely depends on chemical drugs, which hasa strong side effect and drug resistance. The effect of clinical treatment was seriouslyaffected. So the exploration of new antitumor drug with high efficiency and low toxicityfrom Chinese herbal and natural medicine increasingly becomes important.In the present paper, water extract was subjected to macroporous resin columnchromatography eluted with water,30%ethanol,60%ethanol and95%ethanol. Thenthe antifungal and antitumor activities were screened and found that the sub-fraction of30%ethanol possessed the best antifungal activities, with the minmum inhibitoryconcentration (MIC) Trichophyton tonsurans3.125-6.25mg/mL, Microsporumgypseum1.563-3.125mg/mL, the minmum fungicidal concentration (MFC)Trichophyton tonsurans6.25-12.5mg/mL, Microsporum gypseum3.125-6.25mg/mLmg/mL. The result of antitumor activities indicated that each component showedactivities in different degrees inhibiting the growth of A549and MCF-7and fraction of 30%ethanol showed the most significant effect. So it was found that fraction of30%ethanol was the effective part for inhibiting fungi and antitumor.In this study,10compounds including four aromatics two flavones, twosesquiterpenoids, one lignans and one sterol were isolated from the fraction of30%ethanol using silica gel column chromategraphy, Sephadex LH-20columnchromatography, HPLC, MCI column chromatography, TLC etc and identified bymeans of UV, IR, MS, NMR and referring to some references. The one new compoundwas named as Dryofragranoside K. And nine known compounds were named as(Z)-3,5,5-trimethyl-4-(3-(3,4,5-trihydroxy-6-(hydroxymethyl)-tetrahydro-2H-pyran-2-yloxy)butylidene)cyclohex-2-enone,(E)-3-(4-hydroxyphenyl)acryic acid, β-sitosterol,3,4-dihydroxy benzaldehyde,(6S,9R)-3-oxo--ionol-9-O-β-D-glucopyranoside,fragranoside A,7,9′;7′,9-diepoxylignans pinoresinol-4,4′-di-O-β-D-galactoside,(7S,8R)-dihydrodehy drodiconiferyl alcohol-9-O-β-D-glucoside, quercetin-3-O-β-D-glucose.The antimicrobial activities of seven known compounds isolated from the part of30%ethanol were screened. It was showed that (7,9;7,9-diep-oxylignanspinoresinol-4,4-di-O-β-D-galcoside)，(E)-3-(4-hydroxyphenyl)acryic acid,(6S,9R)-3-oxo--ionol-9-O-β-D-glucopyranoside had presented the obvious activitiesinhibiting the growth of Trichophyton tonsurans and Microsporum gypseum.Nine compounds were used to screen antitumor activity by MTT method in vitro.The results showed that the high active for inhibiting A549cells were fragranoside Aand (E)-3-(4-hydroxyphenyl)acryic acid. IC50values were18.75±0.22μmol/L,11.03±0.15μmol/L,8.31±0.26μmol/L and32.78±0.13μmol/L,22.97±0.24μmol/L,22.74±0.18μmol/L in24,48and72hours’,respectively. The high active for inhibitingMCF-7was (7,9;7,9-diep-oxylignans pinoresinol-4,4-di-O-β-D-galcoside),Dryofragranoside K and quercetin-3-O-β-D-glucose. IC50values were17.58±0.13μmol/L,9.52±0.11μmol/L,6.27±0.21μmol/L and27.38±0.12μmol/L,21.50±0.18μmol/L,15.97±0.25μmol/L and30.50±0.15μmol/L,26.46±0.20μmol/L,20.78±0.28 μmol/L in24,48and72hours’, respectively. All showed a certain amount of time andconcentration dependence.In the paper, antimicrobial and anti-tumor activities of fraction of D.fragrans andmonomeric compounds were evaluated. In summary, it was exhibited that the leadingcompounds which provided new medicinal resources for the treatment of skin diseasesinduced by Trichophyton tonsurans and Microsporum gypseum and for the developmentof antitumor drugs. |
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