| The root of Aster tataricus,which is spicy,bitter,warm,and belongs to lung meridian,is one of the most commonly used traditional Chinese medicine(TCM)with the function of moistening lung,eliminating phlegm,relieving cough and asthma for more than two thousand years in China.It was reported that there were kinds of ingredients contained in it.Up to now,almost 60 components were isolated from it.Meanwhile,the results of modern pharmacological researches also indicate that it also has many potential biological activities especially in some aspects of the treatment of respiratory diseases.In recent years,with the deepening of research on it,the advantages of it in treatment of lung injury and asthma have been noticed.For example,it was reported that the p-hydroxybenzoic acid isolated from it could effectively alleviated the lung injury induced by seawater;and what’s more the dual-luciferase reporting system suggested that the extract of it also showed significant activity in asthma.Therefore,on the basis of the previous researches and the traditional medicinal efficacy of it,systematic investigations were established in this study for uncovering the total chemical components composition of it,the correlation between the main active components and the efficacy mentioned above and the mechanisms how components work.And the main content of this study is as follows:(1)As we all know,the efficacy of one traditional Chinese medicine does not only rely on one single component in it,but usually depend on the combined effects of all the active components in it.Therefore,it is very necessary to clarify the chemical constituents of one traditional Chinese medicine for its pharmacological effects and clinical application,and also for the modernization of traditiaonal Chinese medicine.In this study,a systematic data acquisition and mining strategy aimed at the traditional Chinese medicine complex system based on ultra high performance liquid chromatography coupled with quadrupole time of flight mass spectrometry(UHPLC-Q-TOF-MS)was establised.And the workflow of this strategy is as follows:to be first,the high resolution mass data are acquired by both data dependent acquisition mode and data-independent acquisition mode.Then a global data mining strategy are employed,including targeted compound finding,non-targeted compound finding and non-targeted compound finding based on targeted compounds.Next,some assistant tools,such as key product ions(KPIs),are applied for compound hunting and identification.In general,a total of 132 compounds were identified or tentatively characterized in Aster tataricus based on the accurate mass measurements within±5 ppm error in this research.Therefore,this research provided not only reliable data support for the comprehensive analysis of the consitituents in Aster tataricus,but also provided an efficient data acquisition and mining strategy to profile the chemical constituents for other traditional Chinese medicine complex system.(2)TCM believes that the lung is the most delicate viscera and not resistant to cold and hot.Usually,exogenous pathogenic factors(such as trauma,pathogenic toxin,etc.)first attack the lung,leading to lung qi closure,stagnation of qi activity and finally to lung disease.And this phenomenon is consistent with the view of modern medicine on the pathogenesis of acute lung injury(ALI).ALI is a common clinical disease characterized by infection,trauma and other factors that promote to a variety of effector cells to release a mount of inflammatory mediators,resulting in uncontrolled progressive lung inflammation.Although advanced investigations have been made in promoting the understanding of it,much of its mechanism remains unknown.Due to its complex pathogenesis,effective durgs to cure this disease are limited and new therapeutic strategies are needed.Therefore,in the second chapter,the lipopolysaccharide(LPS)was used to establish the model of AIL to evaluate the efficacy of Aster tataricus on moistening lung and descending qi.And then the material basis and the possible mechanism of it were also explored.Firstly,the xylene-induced mouse ear edema model was applied to preliminary screen out the anti-inflammatory fraction of Aster tataricus extract.Then,the LPS-induced acute lung injury model was then carried out to further evaluate its protective effects.After that,the major constituents in it were subsequently identified and the candidate components were selected for further network pharmacology investigations.The experiment results indicated that Fraction-75,which eluted from Aster tataricus extract,could not only significantly inhibit the xylene-induced mouse ear edema,but could also protect the LPS-induced acute lung injury from multiple aspects,including alleviating the severity of lung pathology,attenuating the pulmonary edema,reducing the release of inflammatory cells.Further network pharmacology investigations showed that 10 candidate components inlucded in Fraction-75 could protect the LPS-induced acute lung injury by inhibiting the release of inflammatory cytokines and promoting the repair of vascular endothelial.This work provided a practical way to elucidate the relevance of the ingredients of Aster tataricus and their therapeutic effects,and also provided valuable methodological guidance for visualizing the interaction of drug and disease,which is beneficial for better understanding the complex mechanism of multi-target therapy.(3)Modern medical research revealed that the mechanism of ALI and asthma are not exactly the same.However,there were also some common characteristics between them,for example no matter in the process of ALI or asthma there was usually full of severe inflammation in respiratory tract and lung.Meanwhile,the TCM theory regarding both the ALI and asthma as lung diseases that caused by the descending lung lost and lung qi ascending counterflow.Hence,on the basis of the second chapter,investigations were also carried out to further explore the efficacy of Aster tataricus on preventing asthma.In general,this chapter was desined mainly to investigate the potential biological activity and the mechanism of Aster extract and its three different elution fractions in treatment of asthma in vitro with the help of isolated tracheal rings of juinea pigs and the tracheal smooth muscle cells of mouse.On this basis,the expectorant and antitussive activities of the most effective fraction,which eluted from Aster extract,were further evaluated so as to confirm whether the selected fraction in vitro is also the main active part in vivo.Finally,the chemical components of the selected fraction were analyzed and the correlation between the pharmacological efficacy and the identified components was explored as well.The experimental results showed that the Fraction-75 eluted from Aster tataricus produced an obviously relaxation effect on tracheal rings pre-contracted by KCl.Meanwhile,it also exhibited a concentration-dependent inhibition activity on CaCl2-,acetylcholine-or histamine-induced tracheal ring contaction and it could significantly reduce the level of Ca2+in tracheal smooth muscle cells as well,indicating that the mechanism might be to block calcium channel and antagonize muscarinic or histamine receptor.Further experimental results demonstrated that Fraction-75 also exhibited the best acitivies on expectorant and antitussive in vivo.And the LC-MS results suggested that the flavonoids,phenolic acids and terpenes in Fraction-75 may take the primary responsibility for the cativites above.In conclusion,this study could not only guide people toward a better understanding of Aster tataricus of its efficacy and possible mechanism on moistening lung and relieving asthma,but could also be useful for new drug development. |
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