| Pterocephalus hookeri (BangZiDuWu in Tibetan), which was included in Chinese Pharmacopoeia (2010 version) and Ministry of Public Health’s Quality Standard (1995 version, Tibetan medicine I) was a commonly used Tibetan medicine.There are "identify", " inspect", "content determination", Ursolic acid and Oleanolic Acid used as reference substance is lack of specificity, and the traditional identification wasy lead the results inaccurate apparently. Pterocephalus hookeri treat Rheumatoid Arthritis (RA) effectively according to Tibetan medicine hospital and research of Pharmacodynamics.,the active ingredients is unknown making its clinical medication effectiveness cannot be widely recognised, limits its further application. Pterocephalus hookeri treat Rheumatoid Arthritis (RA) effectively according to Tibetan medicine hospital and research of Pharmacodynamics.,the active ingredients is unknown making its clinical medication effectiveness cannot be widely recognised, limits its further application.Pterocephalus hookeri is perennial herb and was included in Level 1 species endangered of Tibetan medicine in 2000. The each parts of Pterocephalus hookeri were used as medicine according to ancient book literatures of Tibetan medicine and Tibetan medicine hospital.However, the whole herb only in Chinese pharmacopoeia (2010 edition),and will Resources destruction caused.At present the research can’t incarnate chemical composition of different parts.there is no gist to confirm officinal parts.It is necessary to establish identification system by DNA and researching constituents migrating to blood.researching different part Fingerprint by UFLC and pertinence between gene and chemical component to establish new model for quality evaluation and resources protection.Objective:1. Identify authenticity o f pterocephalus hookeri.2. Establish underground and overground fingerprint of pterocephalus hookeri and compare their differences.3.Preliminary research constituents migrating to blood of Pterocephali Herba, lay the foundation for tracking active ingredients.Research pertinence between gene and chemical component.Methods:1. Extracting DNA by Reagent Kit method,PCR amplification the ITS2 sequence,MEGA5.1 software for data analysis.2.UFLC-PDA was adopted to analyse Fingerprint of different parts of Pterocephali Herba.SPSS and Grey relation software for data analysis.3.Researching constituents migrating to blood of Pterocephali Herba Preliminary by HPLC.Results:1.The similarity between the download GenBanksequences and the experimental ITS2 sequences of 57 batch samples of Pterocephali Herba amount to 99%;ITS2sequence contain 232 bp and 6 haplotypes,genetic distance is 0-0.04003,the average of genetic distance is0.0038;57 batch samples of Pterocephali Herba hold together.and approval rating Of Bootstrap over86% according to NJ tree.2.Established underground and overground fingerprint of pterocephalus hookeri.15 (13)common peaks were extracted from overground (underground) Fingerprint,peak 6 and 7 are not exist in overground Fingerprint, Seven peaks which exist in all the parts were identified in the Fingerprint combined with existing reference named chlorogenic acid, loganin.Alpine radix dipsaci glycosides i sweroside, evodia rutaecarpa glycosides, Dipsanoside Band triplostoside A.3. There are low dependency between gene and chemical component except for peaks 4,8,10 and 13(>0.8),Three peaks were identified named loganin, evodia rutaecarpa glycosides, Alpine radix dipsaci glycosides i and triplostoside A.4.There are 9 constituents in blood.5 Prototype components named chlorogenic acid, sweroside.Alpine radix dipsaci glycosides i,evodia rutaecarpa glycosides and Dipsanoside B through comparing chromatogram of Pterocephalus hookeri,drug serum and blank serum.Conclusion:1. ITS2 sequence distinguish Pterocephali Herba steadily and accurately is a new technological means.2.The compounds in overground is More than underground,the overground parts can replace the whole grass in clinical practice theoretically.3.The genetic gene might has great impact on the secondary metabolites of iridoid glycosides.4.Preliminary reveals constituents migrating to blood,lay the foundation for the research of efficacy material. |
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