Preliminary Research On The Material Basis And Action Mechanism Of Chaihu Qingwen Granules In The Treatment Of Acute Lung Injury

Background:As prevalent acute respiratory condition in clinical practice,acute lung injury has a quick start and severe symptoms which can harm patients physically.Chaihu Qingwen granules,a combination preparation based on the decoctions of Xiaochaihu,Guizhi Houpu Xingzi,and Qiwei Anti-Virulent,can be used to treat respiratory conditions such as the common colds,the flu,fever,coughs,and other respiratory illnesses.However,the effect of Chaihu Qingwen granules on acute lung injury has not been reported in the literature.Objectives:The purpose of this study was to investigate the anti-inflammatory effect of Chaihu Qingwen granules on lipopolysaccharide-induced acute lung injury in rats and its potential mechanism,and to explore its pharmacodynamic material basis.Methods:1.The pathological changes of rat lung tissue were observed by hematoxylin-eosin staining.The expression levels of inflammatory factors(IL-17,IL-1β and TNF-α)in serum and bronchoalveolar lavage fluid of rats were determined by ELISA.The expression levels of inflammatory-related proteins in the TLR4/NF-κB/NLRP3 signaling pathway in rat lung tissue were determined by Western blotting and immunohistochemical staining.2.A chromatographic and mass spectrometric method was developed for the qualitative analysis of Chaihu Qingwen granules.Then,using UHPLC-Q-TOF-MS to detect the Chaihu Qingwen granules sample and the control product.The original LC-MS data were processed by the Peak View workstation.According to the primary and secondary fragmentation ion peaks,and consult the relevant literature to identify its chemical structure.Results:1.The pretreatment with Chaihu Qingwen granules can significantly ameliorate lung tissue pathological injury in lipopolysaccharide-induced acute lung injury rats,reduce the release of inflammatory cytokines in serum and bronchoalveolar lavage fluid,which indicated that Chaihu Qingwen granules can inhibit the inflammatory response in rats with acute lung injury.The results of Western blot showed that the expression levels of TLR4 and NLRP3 were down-regulated,while the phosphorylation levels of NF-κB p65 and IκBα were suppressed in lung tissues of rats pretreated with Chaihu Qingwen granules.The results of the immunohistochemical analysis showed that the expression levels of NF-κB p65 and NLRP3 were significantly down-regulated in each dose group of Chaihu Qingwen granules,and the expression level of TLR4 was significantly decreased in the high dose group.2.By referring to the retained time,molecular mass and fragment ion information of existing standards,combined with the database and published literature information,a total of 48 compounds were identified in this study in positive and negative ion mode,including 15 flavonoids,12 organic acids,5 lignans,4 iridoids,4 phenylethanoid glycosides and 8 other compounds.Conclusions:1.The pretreatment of Chaihu Qingwen granules has a strong protective effect on acute lung injury in rats.It is speculated that the protective mechanism of Chaihu Qingwen granules may be related to the inhibition of TLR4 / NF-κB / NLRP3 signaling pathway.2.In this study,a total of 48 chemical components in Chaihu Qingwen granules were identified,most of which have been proved to have anti-inflammatory effects,and a variety of active components can exert anti-inflammatory effects by inhibiting NF-κB signaling pathway,which provides a certain material basis for studying the mechanism of Chaihu Qingwen granules against acute lung injury.