| Asthma, as defined in2008by the Global Initiative for Asthma (GINA), is an inflammatory disorder of the airways in which many cells and cellular elements play roles. Bronchial hyperactivity associates with inflammation that together with an external or environmental insult, on vulnerable bronchial epithelial structures, generates tissue remodelling and respiratory functional impairment. Asthma is not a curable disease at the present time. However, with proper treatments, the risk of mortality for asthmatic individuals could be comparable to that of the general population. Presently, the treatment of asthma includes a dual focus:the short-term treatment of acute symptoms with bronchodilators, and together with the prevention or eventual reversal of chronic inflammation using anti-inflammatory drugs.Qingfei Xiaoyan Wan (QFXY) is originated from a famous Traditional Chinese Medicine (TCM) formula "Maxing Shigan Decoction". It has been experimentally improved, consisting of eight materia medicas, Ephedra Herba, Saigae Tataricae Cornu, Pheretima, Arctii Fructus, Lepidii Semen, Bovis Calculus Artifactus, Armeniacae Semen Amarum and Gypsum Fibrosum. Since decades of extensive clinical practice, QFXY has shown significantly therapeutic effects on dissolving phlegm as well as relieving cough, asthma, upper respiratory tract infection, bronchitis, pneumonia, and etc. Comparing with other anti-asthma drugs, it is characterised with moderate and persistent efficacy as well as few side effects, however, the underlying action mechanism still remains elusive.We first established asthma model induced by histamine phosphate and acetylcholine chloride (His&Ach) in guinea pigs, which then were administered orally with QFXY (132.5mg/100g). Asthma prolonged time and Hematoxylin-Eosin staining sections were applied for evaluating QFXY effect. Comparing with the model group, in the QFXY group, the asthma onset time was significantly prolonged and HE sections presented anti-inflammation and anti-remodelling effects of QFXY. In both Model and QFXY groups, customized microarrays and2D electrophoresis were adopted to detect differentially expressed genes (diff genes) and proteins (diff proteins) respectively, and some diff proteins were identified with MALDI-TOF/MS. As a result,55diff genes (27human homologs after Blasting) and6diff proteins were identified in QFXY group. Validation by qPCR and Western blot showed the microarray and2D data reliable. The checked diff genes and proteins underwent Cluster, GO and KEGG analysis, which focused on signaling pathways in cytoskeleton and extracellular matrix regulation, smooth muscle contraction as well as inflammation response,etc. Based on GAD and HPRD databases, QFXY-asthma target regulation network was constructed, containing1214nodes and1886interactions.Based on the combination of Chemomics and Systems biology research, after the formula study, we next focused on Arctigenin, an active ingredient in QFXY, is reported with anti-inflammation, antioxidation and vasodilator effects. However, its effects on airway smooth muscle and intracellular calcium, as well as ion channels remain eclusive.To investigate how arctigenin regulates bronchus tone and calcium ion (Ca2+) flow. Trachea strips of guinea pigs were prepared for testing relaxation effect of arctigenin (0.1μM-1mM) to10μM acetylcholine,10μM histamine,10mM KCl and30mM CaCl2, respectively. Furthermore, L-type calcium channel currents were detected by patch-clamp and intracellular Ca2+concentration treated with arctigenin in human bronchial smooth muscle cells (HBSMC) was detected by confocal microscopy. The results showed that arctigenin exhibited relaxation effect on tracheae to different constrictors, and this was related to decreasing cytoplasmic Ca2+concentration by inhibiting Ca2+influx partly through L-type calcium channel as well as promoting Ca2+efflux.Hereby, in the first part of the study, a primarily combined genomic and proteomic screening of QFXY targets displayed a series of candidate genes and proteins, which indicated that the effect of QFXY relied on the combined mechanism, anti-inflammation and anti-remodelling, as well as influencing signal transduction in vivo, demonstrating QFXY multi-target network regulation as an asthma controller. ASM related study provides new insight into the mechanisms by which arctigenin exhibits relaxation effect on bronchus and suggests its potential use for airway disease therapy. All together, these results reveal multiple active components, multiple targets and synergistic effects in the mechanism of action of a TCM. |
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