| Cordyceps is a precious traditional Chinese medicine with significant effect and unique sources.It has a long medicinal history in China with the effect of Tonifying kidney and lung, stoping bleeding phlegm,mainly used in the treatment of chronic cough asthma, impotence, spermatorrhea,Labor cough hemoptysis, lumbar debility, the lung or kidney disease. the18th century （1785）, when British mycologists Dickson brought Cordyceps back to western countries for the first time, this traditional medicine with magic began to be known by the whole world, which also opened the door of modern research about Cordyceps. The modern pharmacology studies have proved that Cordyceps sinensis is rich in a variety of chemical ingredients. Now, its tonic effect mainly clinically manifested of immune regulation with unique effect.With more and more attention to the sub-health, Cordyceps as the tonic has an increasing market deman for its mild nature.But, its annual production is limited,and alternatives such as its artificial cultivation research is still in its crucial stage. Its expensive price, increased year by year. A lot of adulteration products, adulterants, have appeared in the market,and its distribution channels is more chaos, not-standard. The definitions of Cordyceps sinensis varieties in The Chinese Pharmacopoeia are open to question. The quality standards of the Chinese Pharmacopoeia. The index components of Cordyceps in The Chinese Pharmacopoeia is the adenosine class composition that is entomogenous fungi widespread, which is difficult to effectively targeted authenticity and quality evaluation.Protein ingredient is an important part of the chemical composition in Cordyceps.Protein itself is the variety expression of genetic information, and usually has a strong biological activity, proteomics research is the focus of world, especially in differences in protein expression and functional protein screening, which has unique advantages,while the study of Cordyceps protein components has been lagging behind, few research. In view of this, the subject of Cordyceps carried out the research work of the following aspects: 1. Species research of CordycepsResearch base of the original varieties of Cordyceps bacteria, parasites, describes the progress of the study of the taxonomy of Cordyceps sinensis. According to the latest progress of molecular studies in2007. the Teleomorph and Maxi has been classfied as Ascomycetes.Linear Cordyceps Branch,Linear Cordyceps Ophiocordyceps sinensis （Berk.） G. H. Sung, J.M.Sung, Hywel-Jones&Spatafora Abbreviated as Ophiocordyceps sinensis （Berk.） G. H. Sung et al. Propose Pharmacopoeia update the base of the original species of Cordyceps. Through the research of theVarieties of the Cordyceps parasites origin,we found that Cordyceps parasite species has59species, while most scholars have recognized that the host insects of Cordyceps is a Hepialus in Hepialidae Hepialus.More systematic study of Chinese medicine Cordyceps naming and analysis of Calendar Chinese Pharmacopoeia and related literature show that "dongchongxiacao" as a medicinal Chinese name is more objective, appropriate, but not specific.Chinese medicine Cordyceps should be different from Cordyceps fungus, Latin name should not be directly equivalent to the generic name of the Cordyceps fungus.Propose naming medicine material via Combination of the insect body parts and latest scientific name of Cordyceps fungus:Ophiocordyceps Hepialus.2.2-DE method optimization and the identification of the characteristic protein of Cordycepson the basis of Task Force preliminary study, This paper further applicable to Cordyceps protein2-DE method, obtained the best method:（1）.the sample volume of600mg;（2）.passive hydration;（3）.build salt bridge to further demineralization.Reproducibility study results showed that the method of the same batch of samples of the same origin match rate reached89.5%, indicating that the method is reliable。Studies on the determination of protein content in Cordyceps2-DE process showed the Bradford method is optimal as the detection method of Cordyceps soluble protein.Through the research of the difference of different origin cordyceps protein expression profiles and intercommunity,12proteins characteristic point was found, and five of them in the structure of the preliminary appraisal.The structure identified: Acetolactate synthase small subunit of class,Transcription repair coupling factor class,Hypothetical proteins Mnod7043class,FMN-dependent alpha-hydroxy acid dehydrogenases,Iron-sulfur oxidoreductase protein.Based on analyzing the diversity of different origin cordyceps （Differences in protein, total protein）., results showed that different regions authentic Cordyceps protein species had difference,combining with the variety in front of the study. Speculated that the diversity of Cordeceps protein and different locality advantages cordyceps fungus host insect species..This laid the foundation for the follow-through the Cordyceps characteristic protein on different Cordyceps Origin Traceability. At the same time put forward by finding common proteins of different sources of authentic traditional Chinese medicine, to evaluate the quality of traditional Chinese medicine, which can be a reference for quality evaluation of the traditional Chinese medicine with many origins as well as complex.3. Immunologically active protein of CordycepsBy enzyme-linked immunosorbent assay （ELISA） determined the influence of drugs on macrophage secretion of TNF-a, IL-1, IL-12factor. Research on Cordyceps protein component of the immune activity to optimize the in vitro immune activity detection method:Isolate mouse peritoneal macrophages, Incubate for3h cells adherent, Dosing of LPS （as a positive control for the post-experiment） dosing a final concentration of10μg/ml,Terminate reaction after dosing9h, discard nutrient medium, replace of Antibiotic-free medium.Test should be in mouse peritoneal macrophages,completed within72h after extraction and purificationConducted a study on the stability of the protein component, screened a suitable protein component storage conditions:Buffer PB （pH=7.2,0.45mmol/L DTT） as the Cordyceps protein preservation solution,-20℃for10dThrough a combination of immune activity tracking, ultrafiltration separation Cordyceps protein, got four molecular weight segment.ultrafiltration has better effect.Immune activity test results showed that the total protein extract of Cordyceps and the molecular weight of the protein components had a wide range of bidirectional regulation to immune system macrophages.（1）.Functional bidirectional adjustment,Which was to that protein of different molecular weight on normal macrophages had certain promotion, inhibition, or effect was not obvious;To the low immune status of the cells （3-mercaptoethanol modeling injury）, it can significantly promoted the secretion of immune factors to near normal levels or enhance immune function;To the over-exuberant immune status of the cells （LPS stimulation）, it can significantly inhibited the secretion of immune factors to near normal levels or had a immunosuppressive trend.（2）. Doses of bidirectional regulation. Samples of different molecular weight protein regulation of immune factors, there was not a significant linear relationship. But on the whole, within a certain concentration,there was a two-way regulation in which high concentration promoted immunization,low concentration had an immunosuppression,（3）.Different molecular weight type of bidirectional adjustment.Although the protein components of different molecular weight immunomodulatory effect of different immune factors appeared to be different, on the whole, analysis found, mainly for immunologic enhancement of molecular weight protein components, with lower molecular weight, immunosuppression more obvious.Preliminary separation of protein components of Cordyceps by gel chromatography. Got protein Ⅲ characterized by immune inhibition, activity strong, good reproducibility, types of relatively single, in14.4～20.1kDa protein molecular weight. Which provided the basis for later high-throughput screening of active protein.The research on Cordyceps characteristic protein and active protein, Namely, from the point of the identification of ingredients and active ingredients, explored research on of Cordyceps quality evaluation.which laid the foundation for the later seeking indicators protein or protein components and provided the scientific basis for Cordyceps quality evaluation from proteomics research... |
PDF Full Text Request