| The traditional Chinese medicine Sanghuang recorded in ancient literature was textually criticized by Herbal textual research in the paper first.On this basis,the traditional efficacy that ‘Sanghuang' could treat women's amenorrhea,blood clotting and abdominal mass were validated by using modern pharmacology method.Meanwhile,the chemical composition,anti-inflammatory and cytotoxicity of Inonotus hispidus were studied.Finally,the high performance liquid chromatography(HPLC)fingerprint and DNA barcodes were established to constitute identification system of Sanghuang.Herbal textual research: Sanghuang is an internationally recognized medicinal fungus with good anticancer and immune enhancement effects;however,different countries and different regions have their own understanding about Sanghuang.Now although there are many scholars devoted to clarify Sanghuang,there is still confusion on medicinal material varieties,homonym,synonyms phenomenon,which do not tally with the records of ancient literature.On the basis of the original research,the supplementary research of Sanghuang including accessing and consulting to a large number of literature and on-the-spot investigation were carried out by the author.The results showed that Inonotus hispidus was truly traditional Chinese medicine Sanghuang recorded in ancient materia medica.I.hispidus former name was Xanthochrous hispidus,belongs to Inonotus,Hymenoehaetaceae,Hymenoehaetales,Agaricomycetes,basidiomycetes Basidiomyeota phylum,is a kind of wood-decay fungi growing in temperate zone.Mainly grows in the mulberry,poplar,elm,ash-tree,walnut tree,maple etc living standing timber and occasionally grows on a fallen tree.During the field resources survey,it was found that I.hispidus was mainly grown on ancient mulberry trees in Aksu area of Xinjiang and Xiajin County of Shandong,and grown on Ulmus macrocarpa var.mongolica in the western area of Jilin province.The validation of traditional efficacy: In order to confirm Sanghuang's sweet and pungent nature,non-toxic,treatment of women in the collapse of the band,closed blood coagulation,postpartum hemagglutination recorded in Yao Xing Lun published in Tang Dynasty by Zhenquan,the basidiocarp of I.hispidus were taken from the mulberry,poplar,elm,ashtree,Ulmus macrocarpa var.mongolica and the effect of blood rheology in syndrome of congealing cold with blood stasis rats was studied.The experimental results showed that I.hispidus basidiocarp with yellow-brown had significant effect on blood rheology of rats with congealing cold and blood stasis,P<0.05.However,the I.hispidus basidiocarp parasitized in the same tree species falling off their corresponding characters in black has no significant influence for the effect of blood rheology in syndrome of congealing cold with blood stasis rats.The experimental results are consistent with the literature.The correlation degree of the Quantitative peak area of 32 characteristic peaks from 6 different sources and the effect of blood rheology in syndrome of congealing cold with blood stasis rats were analyzed by the grey system theory modeling software(GSTAV 7.0).The results indicated that the peaks with retention times of 73,75,79,81,90,94,98,100,146,161,167 under the chromatographic condition described in this paper were closely related to the changes of plasma viscosity.The chemical composition research: In the paper,the chemical composition of I.hispidus' mature yellow basidiocarp were researched.Systematic separation and purification were proceeded to study its chemical composition,and five polyphenolic compounds were separated from the basidiocarp of I.hispidus including phellibaumin A,phelligridin C and phelligridin D,3'4'-dihydroxy-5-[11-hydroxyphenyl]-6,7-vinyl]-3,5-dioxa-fluoren-5-one,respectively.Of which,3'4'-dihydroxy-5-[11-hydroxyphenyl]-6,7-vinyl]-3,5-dioxa-fluoren-5-one was a new compounds.It was found that phelligridin C was not a single compound,but cis-trans isomers composed of phelligridin C mixture.cis isomer of phelligridin C is also new compound.Phellibaumin A,phelligridin C and phelligridin D were firstly isolated from I.hispidus.At the same time,the compounds phellibaumin A,phelligridin D,3'4'-dihydroxy-5-[11-hydroxyphenyl]-6,7-vinyl]-3,5-dioxa-fluoren-5-one were used for anti-inflammatory activity test and cell toxicity test.The results of anti-inflammatory activity test showed that all the three compounds had anti-inflammatory activity.The compound3'4'-dihydroxy-5-[11-hydroxyphenyl]-6,7-vinyl]-3 had relatively stronger anti-inflammatory activity.Cytotoxicity results showed that the three compounds had little effect on cell proliferation and had high biosafety.The establishment of HPLC fingerprint and DNA barcodes: As the types and relative proportions of the chemical constituents contained in the I.hispidus were related to pharmacological activities,the fingerprint of I.hispidus was tried to established by using HPLC and the content of indexity was determined as a reference for its quality measurement.In the experiment,it was found that the distribution and proportion of the chromatogram peaks were different from the I.hispidus solution liquid chromatogram parasitized on different trees,and the unified fingerprint could not be established.Due to the limit of sample collection,the I.hispidus parasitized Ulmus macrocarpa var.mongolica was chosen as the object in this study.The fingerprint of I.hispidus parasitized Ulmus macrocarpa var.mongolica was established by HPLC.The determination method of phelligridin D content was established with the self-made phelligridin D as reference substance.The results showed that the fingerprint method was stable and feasible.It could effectively identified I.hispidus parasitized Ulmus macrocarpa var.mongolica.The result showed that the content of phelligridin D of I.hispidus parasitized Ulmus macrocarpa var.mongolica is 1.0-1.2mg/g.In order to identify different species and different sources of Sanghuang fungi,a relatively conservative ITS2 fragment was used as identification means.The experimental results showed that the system phylogenetic trees constructed by NJ and MP were all similar.Different Sanghuang fungi were all in the systematic phylogenetic tree and different branches,of which,NJ method used the shortest time,and MP method used longer time.However,all methods confirmed that ITS2 fragment could effectively distinguish different Sanghuang fungi,so ITS2 fragment can be used as the ideal DNA barcode for distinguishing Sanghuang fungi resources.This paper is of great significance to the deep research,development and utilization of traditional Chinese medicine---Sanghuang. |
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