Study On The Function And Active Constituents Of Zhikechuanbeipipa Dropping Pills And Weichangan Pills

Zhikechuanbeipipa Dropping Pills（ZKC）,a traditional Chinese medicine prescription,has been widely used to treat chronic obstructive lung disease and cough caused by bronchial catarrh.Weichangan Pills（WCA）has been used to treat irritable bowel syndrome and functional diarrhea effectively.The study was aimed to evaluate the bronchodilatory activity of ZKC and the potential mechanisms involved.To further elucidate the potential mechanisms of the action involved,we investigated the bronchodilatory effects of Folium Eriobotryae（FE）,the main herb in ZKC,and its major active components,ursolic acid（UA）.Guinea pig models of isolated trachea in vitro and acetylcholine（Ach）and histamine（His）-induced asthma in vivo were used to investigate the anti-asthmatic effect of ZKC.The results showed that ZKC（2.5,5,10 mg/mL）significantly inhibited Ach and His-induced trachea contractions and ZKC（1.78,3.55,7.1 mg/kg）prolonged the latent period of asthma by Ach and His in guinea pigs.In in vitro and in vivo screening experiments,ethyl acetate fraction of FE（EFE）showed the most active relaxing effect.To determine the chemical composition of EFE,UPLC-Q-TOF/MS was used to separate and identify the constituents.Based on the retention time,accurate mass and characteristic MS² fragments,a total of 35compounds,including 18 triterpene acids,10 flavonoids,5 phenolic acids and 2 fatty acids,were identified or assigned tentatively by comparing with known compounds or consulting the respective literatures.UA accounted for 10%of EFE.The effects of EFE（0.05-0.3 mg/mL）on the isolated tracheal strips were tested in vitro.Meanwhile,Ach and His-induced bronchospasm were conducted in vivo.Furthermore,mechanisms of relaxant effects of EFE were evaluated in the absence and presence of specific inhibitors.With in vitro studies,the contractile response evoked by Ach or His(EC₅₀=0.21 and 0.16mg/mL)was decreased by EFE.With in vivo studies,EFE strongly prolonged preconvulsive time similar to isoprenalin.The bronchodilator effects of EFE could be blocked by propranolol（1μM）,NG-nitro-L-argininemethylester（L-NAME）（100μM）and1H-[1,2,4]oxadiazolo[4,3-a]quinoxaline-1-one（ODQ）（1μM）.EFE also inhibited the contraction in Ca²⁺-free medium and produced rightward parallel displacement of CaCl₂ curves.These results suggest that bronchodilator activities of EFE were related to activation ofβ-adrenoceptor and NO/cGMP pathway.Blockage of Ca²⁺channels and inhibition of IP₃R-mediated internal Ca²⁺release were also involved.The effects of UA（5-30μg/mL）on the isolated tracheal strips were tested in vitro.Meanwhile,ovalbumin（OVA）-induced bronchospasm were conducted in vivo.Furthermore,mechanisms of relaxant effects of UA were evaluated in the absence and presence of specific inhibitors.In in vivo study,UA could decrease the contents of ET-l or NO and adjust the level of IL-5 or INF-γin the BALF.The results showed that UA produced relaxant effects by the NO/cGMP pathway and blockage of Ca²⁺channels.Moreover,we also investigated the protective effects of WCA on non-steroidal anti-inflammatory drugs（NSAIDs）in this study.The results showed that WCA（0.1,02,0.4 g/kg）could significantly reduce gastric ulcer index,and effectively retard gastric mucosal damage caused by aspirin and diclofenac,characterized by improving the mucosal edema,hemorrhage and erosion.In conclusion,all these researches provide a theoretical support for the scientificity of the prescription,and supply the basis for the clinical application and further development of Zhikechuanbeipipa Dropping Pills and Weichangan Pills.