cimifugin and 7-3--dihydroxy-4--methoxyisoflavone

Long-term exposure to aeroallergens such as house dust mite (HDM) could result in airway inflammation and airway remodeling, characteristic features of allergic asthma. Huangqi-Fangfeng (HF), an important "couplet medicines" of Yu-Ping-Feng-San (YPFS), mediates allergen-induced airway inflammation in mice, but its role in the airway remodeling is not known.. To evaluate the effects of HF on airway remodeling of allergic asthma in a murine model and to investigate the underlying mechanisms in vivo and in vitro.. The main components of HF were analyzed by HPLC. The HDM-induced asthma mice model was established to study the effects of HF on airway inflammation and airway remodeling in vivo. Enhanced pause (Penh) index value was used as an indicator of airway hyper-reactivity. Bronchoalveolar lavage fluid (BALF) was processed for differential cell counting and determination of cytokines production. The lungs were fixed in 4% paraformaldehyde for histological examination after staining with H&E, trichrome and IHC. Production of interleukin (IL)-4, IL-5, IL-13, and transforming growth factor beta-1 (TGF-β1) in BALF and lung tissues, IgE in serum were measured by ELISAs. Expression of epithelial markers and mesenchymal markers were detected by immunohistochemistry and western blots. The effects of HF and its components on epithelial-mesenchymal transition (EMT) were detected in human bronchial epithelial cells (16HBE) treated with TGF-β1 and HDM.. The main components of Huangqi-Fangfeng detected by HPLC were Calycosin, Formononetin and Cimifugin. In HDM-induced allergic asthma mice model, respiratory exposure to HDM lead to airway hyperresponsiveness and thickening of the smooth muscle layer in the airway. TGF-β1 levels increased in mice airways while epithelial cells lost expression of E-cadherin and gained expression of the mesenchymal proteins N-cadherin, α-SMA and collagen І. These changes were relieved by treatment with HF. Furthermore, restored epithelial markers expression treated with individual components were also detectable in 16HBE cells.. These results demonstrated that Huangqi-Fangfeng protected against allergic airway remodeling through inhibiting epithelial-mesenchymal transition process in mice via regulating epithelial derived TGF-β1.

Topics: Airway Remodeling; Animals; Anti-Asthmatic Agents; Apiaceae; Asthma; Astragalus propinquus; Bronchi; Bronchoalveolar Lavage Fluid; Chromones; Disease Models, Animal; Drugs, Chinese Herbal; Epithelial Cells; Epithelial-Mesenchymal Transition; Humans; Isoflavones; Lung; Male; Mice, Inbred BALB C; Transforming Growth Factor beta1

Yu Ping Feng San (YPFS, in Chinese: Jade Windscreen Powder), a well-known traditional Chinese medicine, is commonly used to cure the diseases of respiratory systems and immune systems.. A selective and sensitive high-performance liquid chromatography coupled with mass spectrometry method (HPLC-MS) was developed and validated for simultaneous quantification of cycloastragenol, formononetin, calycosin, 4'-O-β-glucopyranosyl-5-O-methylvisamminol (GMV) and cimifugin in rat plasma after oral administration of Yu Ping Feng San decoction.. Plasma samples were extracted via solid-phase extraction (SPE), separated on a Zorbax SB-C18 column, detected by single quadruple mass spectrometry with an electrospray ionization interface, and quantified using selected ion monitoring mode. The current SPE-HPLC-MS assay was validated for linearity, intra-day and inter-day precisions, accuracy, extraction recovery and stability. The method was applied to a comparative pharmacokinetic study after administration of Yu Ping Feng San to rats at different doses (10, 20 and 40g/kg).. The calibration curves were linear over the ranges 0.50-50ng/mL and 17.36-1736ng/mL. Intra- and inter-day precisions (relative standard deviations) were from 0.45% to 10.95%, and accuracy (relative recovery) from 95% to 115%. The extraction recoveries were greater than 88.42% for all analytes. Dose-dependence was shown for some constituents in the drug concentration-time profiles. Among all the active ingredients detected, cimifugin had the highest blood concentration (881-1510ng/mL), and cycloastragenol had the longest retention time in the rat body (15.06-20.44h).. This analytical method is a selective, sensitive, precise, accurate, and reliable assay for simultaneous determination of cycloastragenol, calycosin, formononetin, GMV, and cimifugin in rat plasma.

Topics: Administration, Oral; Animals; Chromatography, High Pressure Liquid; Chromones; Drugs, Chinese Herbal; Isoflavones; Male; Mass Spectrometry; Rats; Rats, Wistar; Reproducibility of Results; Sapogenins; Solid Phase Extraction