| Objective:as a common disease,acute lung injury（ALI）causes serious harm in various diseases.In terms of clinical treatment and cure,the existing methods are mostly high-dose oxygen or treatment with dexamethasone and other hormones.Moreover,due to its acute condition,rapid onset process and a series of complications such as pulmonary edema,Therefore,the need for a new way and method becomes particularly critical.As a traditional medicine,traditional Chinese medicine prescription and traditional Chinese medicine mixture have certain outstanding effects in the treatment of diseases,In this experiment,the traditional Chinese medicine prescription has a certain effect of clearing lung and resolving phlegm in the clinical experiment.Therefore,this experiment aims to carry out quality control,extraction and preparation of Qinbai Qingfei mixture on the premise of having a certain clinical basis.At the same time,it uses the means of data analysis to predict the key pathways and later pharmacodynamic research.method;Firstly,according to the provisions in the edition of Chinese pharmacopoeia in 2020,the water extraction of Qinbai Qingfei mixture was carried out,and the orthogonal optimization design was carried out.The investigation factors were water addition,decoction time and extraction times.The best process was selected according to the score according to the extract rate and the content of baicalin,and then filtered through 200 mesh screen,The preservation time is increased by adding preservatives,and then the gradient concentration is carried out,and the molding process is obtained according to the data analysis.Finally,the filling is carried out,and then the subsequent experiments are carried out.Then,in the experiment of quality control,the TLC identification of five traditional Chinese medicines of Scutellaria baicalensis,winter flower,Trichosanthes,Forsythia suspensa and Chelidonium Chelidonium was established,and then the stability,repeatability,sample recovery and precision were tested.The liquid phase determination method of baicalin content in Qinbai Qingfei mixture was determined,and then the standards of microorganism and pH were investigated.Then,according to the data analysis of network pharmacology,through the analysis and screening of the therapeutic targets and action targets of each traditional Chinese medicine in the traditional Chinese medicine formula,through the composition and data mining to find the potential therapeutic targets,and screen the key therapeutic pathways for subsequent experiments.Finally,the acute lung injury model induced by intratracheal instillation was used for subsequent pharmacodynamic analysis.The rats were divided into 6 groups:blank group（KB）,model group（m）,dexamethasone group（DM）,high dose group（H）,medium dose group（MID）and low dose group（L）.After two weeks of preventive administration,the film was made for 24 hours,According to the general activity state of rats,lung permeability,changes of total number of cells and inflammatory factors in BALF（alveolar lavage fluid）,pathological changes and scores of lung tissue,indicators in abdominal aortic blood,blood gas analysis,indicators of oxidative stress in lung tissue,etc.,at the same time,the results of data mining are verified by Western blot,and finally reasonable results are obtained.result;The preparation process of Qinbai Qingfei mixture is to add 14 times the amount of water,the extraction time is 1H,the extraction times is 3 times,combine the Decoction Extracted by water,pass the 200 mesh sieve at room temperature,filter the residue,let it stand and cool,take the supernatant for concentration,concentrate to 100ml,add the preservative sorbic acid 0.2%,mix well with the flavoring agent sucrose,adjust the pH to 4.5～5.5,and repack it by machine.Scutellaria baicalensis and winter flower are determined in the quality control,TLC identification of five traditional Chinese medicines:Trichosanthes,Forsythia suspensa and Chelidonium Chelidonium.Baicalin was determined by HPLC.The regression equation of baicalin was a=4E+07C-76287,R2=0.9996,the linear range was 0.017～0.272mg/ml,the recovery was 99.212%,and the RSD was 0.41%.The established measurement method is feasible,and then the prediction of network pharmacological targets is carried out.According to the data analysis,it is considered that TNF@and NF KB are the key pathways of Qinbai Qingfei mixture in the treatment of acute lung injury（ALI）,while MPO,SOD,GSH-Px,IL-1B,IL-6 and other targets are also the key targets.It is also verified in the subsequent Western blot experiment that the target protein increases with the increase of the concentration of Qinbai Qingfei mixtureThinning has a certain therapeutic effect,which verifies the verification of network pharmacology.In the pharmacodynamic study（1）through the detection of the concentration of permeability BCA protein on the lung surface and the dry and wet weight of the lung,it is found that compared with the model group（m）,the high-dose group（H）of Qinbai Qingfei mixture can significantly reduce the degree of pulmonary edema,Reduce lung permeability（P<0.05）（2）at the same time,by detecting the total number of cells in BALF,it is found that compared with the model group（m）,the high-dose group（H）of Qinbai Qingfei mixture can slow down the progress of the disease and strive for more treatment time if there is subsequent treatment（3）changes in blood gas analysis of rats,Compared with the model group（m）,it was found that the high dose group（H）of Qinbai Qingfei mixture could reduce the decrease of oxygenation index（P<0.05）（4）the inflammatory factors of TNF@,IL-1B and IL-6 in BALF（alveolar lavage fluid）were detected by ELISA,It was found that compared with the model group（m）,the high-dose group（H）of Qinbai Qingfei mixture had a significant effect on reducing the production of these three inflammatory factors（P<0.05）（5）through the detection of SOD and GSH-Px,it was found that the high-dose Qinbai Qingfei mixture（H）could significantly improve the antioxidant enzymes in the lung,Reduce lung tissue injury（P<0.05）（6）according to microscopic HE staining and lung tissue injury score,it can also be found that the high-dose group of Qinbai Qingfei mixture has a good effect on inhibiting the occurrence and progress of the disease.Conclusion;The preparation process and method of Qinbai Qingfei mixture,which is simple and easy to defecate,are determined,and the process is reasonable,low cost and easy to operate.The quality evaluation standard and the determination method of high-phase liquid phase are formulated.A new data mining method of network pharmacology is used to predict the therapeutic targets,which is verified by experiments in the later stage.It is found that Qinbai Qingfei mixture can affect the occurrence and development of Ali（acute lung injury）disease by affecting NF-κB and TNF@signaling pathways.Finally,the effect of traditional Chinese medicine mixture was evaluated by means of pharmacodynamics.It was found that Qinbai Qingfei mixture can reduce LPS（lipopolysaccharide）-induced Ali（acute lung injury）by affecting oxidative stress and inflammatory response. |
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