Study On The Difference Evaluation And Spectrum-effect Relationships Of Gaoben Medicinal Metarials In The Mainstream Markets

The dried rhizomes and roots of Ligusticum sinense Oliv.and Ligusticum jeholense Nakai et Kitag.are traditional Chinese medicine named Gaoben.They have the effect of dispelling wind,dispersing cold,removing dampness and relieving pain.The dried rhizomes and roots of Conioselinum vaginatum(Spreng.)Thell,named Xinjiang Gaoben,entered the local medicine market in 1960s and 1970s.It is similar to Gaoben in appearance and morphology.Because people think that it has the similar effect with Gaoben,it is often sold as Gaoben in the market.Gaoben medicinal metarials mainly contains phthalein,terpenoids,organic acids and other chemical components.Modern research shows that it has cardio-cerebrovascular protection,neuralprotection,anti-inflammatory,analgesic activities and other effects.At present,the quality control of Gaoben in Chinese Pharmacopeia(2015)is mainly focused on the content determination of ferulic acid.And it is difficult to really reflect the overall quality of Gaoben.The unclear potential active constituents also make the quality evaluation system defective.It is also difficult to ensure the safety and effectiveness of alternative use of Xinjiang Gaoben.Therefore,it is necessary to improve the quality evaluation system of Gaoben medicinal metarials,to evaluate the difference and study the spectrum effect relationship of Gaoben medicinal metarials.This dissertation consists of the following four parts:1.Study on the chloroplast genome of Gaoben medicinal metarials.The results show that the chloroplast genome of L.sinense,L.jeholense and C.vaginatum are all typical tetrad structure.In terms of length,GC content,IR boundary,codon preference and repeat sequence,L.sinense,L.jeholense are similar to each other to a certain extent,while C.vaginatum is different from them.The phylogenetic analysis showed that L.sinense and L.jeholense are closely related and clustered into one branch.C.vaginatum is slightly farther from L.sinense and L.jeholense.The results of slide window analysis showed that trnH gene is inserted between ycf2 gene and trnL gene in C.vaginatum,,while trnH gene in L.sinense and L.jeholense is not inserted in ycf2-trnL,but between psbA gene and trnL gene.There are differences in accD-ycf4 fragment between L.sinense and L.jeholense.The above fragments provide new means for the rapid and effective differentiation of the three species.2.Chemcial analytical methods of Gaoben medicinal metarials by UPC2-PDA-QDA and UPLC-PDA-QDA are established.And the contents of butylidenephthalide,Z-ligustilide,senkyunolide A,senkyunolide C,ferulic acid,coniferyl ferulate,D-tryptophan,chlorogenic acid,cryptochlorogenic acid,isochlorogenic acid B,senkyunolide I,butylphthalide are determined.The results show that there are some differences in the types and contents of chemical constituent amomg Gaoben medicinal metarials,for example,the content of senkyunolide A is higher in L.sinense,but lower in L.jeholense and C.vaginatum,the content of Z-ligustilide is higher in L.jeholense,but lower in Z.sinens and C.vaginatum,the content of most of the components are generally lower in C.vaginatum,the components such as butylidenephthalide,butylphthalide and cryptochlorogenic acid even cannot be detected in C.vaginatum.The results of comparative analysis between UPC2-PDA-QDA and UPLC-PDA-QDA show that UPC2 and UPLC are suitable for the analysis of different compounds respctively.Butylidenephthalide,Z-ligustilid,senkyunolide A and senkyunolide C analyzed by UPC2 are all phthalides,which exhibit potential biological activities,such as anti-inflammatory,analgesic activities,cardio cerebral vessels protection and neuralprotection.Chlorogenic acid,cryptochlorogenic acid and isochlorogenic acid B analyzed by UPLC are organic acids,which are widespread in many medicinal materials.The results of chemometric analysis show that UPC2 method has a good identification ability for different Gaoben medicinal metarials.The compounds analyzed by UPC2 method have important chemical taxonomic significance and are more suitable for the chemical quality evaluation of Gaoben medicinal metarials.Moreover,UPC2 is an efficient,economical and eco-friendly method.All the above results show that UPC2 and UPLC methods display good complementarity.UPC2 method exhibits a more effective analysis method for volatile oil rich medicinal materials,and provide a strong supplement for the current quality evaluation system.3.Study on the anti-inflammatory and analgesic activity of Gaoben medicinal metarials.The results show that L.sinense,L.jeholense and C.vaginatum methanol extracts have significant anti-inflammatory activity,which can significantly reduce the contents of NO,IL-1β,IL-6 and PGE-2 in the inflammatory model.The anti-inflammatory activity of C.vaginatum is weaker than that of L.sinense and L.jeholense in the same dose.L.sinense,L.jeholense and C.vaginatum methanol extracts can significantly reduce the times of writhing response induced by acetic acid,improve the pain threshold of thermal stimulation in mice,and have obvious analgesic effect.The analgesic effect of C.vaginatum in low dose(0.05 g/kg)is better than that of L.sinense and L.jeholens.The analgesic effect of C.vaginatum in high dose(0.1,1 g/kg)is weaker than that of L.sinense and L.jeholens.4.Study on the spectrum-effect relationships of Gaoben medicinal metarial s.The results show that there is a significant correlation between chemical co mponents and pharmacodynamic indexes.Compounds 4(senkyunolide A),5,6,7,12(ferulic acid),14(coniferyl ferulate)analyzed by UPC2 are positively c orrelated with the anti-inflammatory activities;compounds 1(butylidenephthalid e),2(ligustilide),3,9,10(senkyunolide C),11,13,14(cryptochlorogenic acid)analyzed by UPC2 are positively correlated with the analgesic activity.Compo unds 1,3(chlorogenic acid),4(cryptochlorogenic acid),6(ferulic acid),7(iso chlorogenic acid B),9,16,18(coniferyl ferulate),19,20(butylphthalide),22 a nalyzed by UPLC are positively correlated with the anti-inflammatory activities,compounds 1,3(chlorogenic acid),4(cryptochlorogenic acid),7(isochlorogen ic acid B),8,10(senkyunolide I),14,18(coniferyl ferulate),22,24 analyzed by UPLC are positively correlated with anti-inflammatory activity.This dissertation focuses on the evaluation of the differences among Gaoben medicinal metarials by chloroplast genome,chemical analysis and pharmacodynamic activity study.Potential biaoactive constituents with anti-inflammatory and analgesic active of Gaoben medicinal metarials were found according to the results of the spectrum-effect relationships.It is necessary to further study on active verification of potential biaoactive constituents and discover their pharmacodynamic mechanism.