| Objective:With the rapid development of emerging information technologies such as big data,cloud computing,Internet of things and artificial intelligence,starting with clinical efficacy evaluation,action mechanism research and standardized application,promoting the modernization and international development of traditional Chinese medicine will become an important way to promote the inheritance of ethnic medicine and the development and utilization of ethnic medicine,and show the advantages of rich medical resources of ethnic minorities.In this study,Kangfuxin Liquid,a traditional Chinese medicine with Yunnan characteristics,was selected as the research object,uses the research method of pharmacoepidemiology to mine and analyze the current situation of clinical medication in the real world,Evaluate the safety and effectiveness of the drug after it is put on the market,predict the active components and possible mechanism of Kangfuxin Liquid in the treatment of ulcer and wound with the help of network pharmacology and molecular docking technology,verify the mechanism of core compounds and targets promoting wound repair through in vitro cell experiment.Methods:1.Using retrospective study method,based on the hospital information system of the inpatient department of the First Affiliated Hospital of Dali University,the use of Kangfuxin solution in the inpatient department was investigated.The two departments with the largest sales volume were selected to extract the historical electronic medical records of drug patients for data sorting and analysis.Set inclusion and exclusion criteria,collect patient admission information,disease diagnosis,course records,historical medical orders,discharge records,etc.,and build a database.Ecological investigation was used to analyze the trend changes of sales volume and number of patients of kangfuxin solution in the past six years,evaluate the safety and effectiveness of drugs after marketing from various aspects.2.By collecting documents and LC-MS analysis to obtain Kangfuxin liquid active ingredient,predict its potential targets,using Gene Cards database to predict disease related targets.Topological analysis and artificial intelligence algorithm are used to predict core targets and core signal pathways,construct active ingredient target pathway network diagram,and explore its possible mechanism.Explore the binding conformation and possible binding sites between small molecule drugs and macromolecular receptors with the help of molecular docking technology.3.In vitro cell experiments were used to verify the possible mechanism suggested by virtual screening.The effect of predicted core compounds on proliferation of CCD841 cell line was detected by MTT assay.The effect of core compounds on migration of CCD841 was investigated by scratch experiment.The m RNA relative expression levels of ADORA2 a and EGFR in CCD841 cell line were detected by RT-QPCR.Results:1.Clinical medication records show that Kangfuxin liquid has been widely used in obstetrics and hematology department of our hospital from January 2015 to December2020,accounting for 41.05% and 14.84% of the total prescribed dosage of Kangfuxin liquid,respectively.It is mainly used for the treatment of oral ulcers caused by chemotherapy in patients with hematologic diseases,and the incision wound caused by perineal lateral resection during childbirth.A total of 2580 patients were included in this study,including 671 patients with chemotherapy-related oral ulcer and 1909 patients with lateral perineal resection.Clinical data show that: in recent six years,the effective rate of Kangfuxin solution in treating chemotherapy-related oral ulcer and promoting wound repair is above 80%,which can effectively shorten the healing time of ulcer and wound,relieve pain,limit the occurrence and development of oral disease and wound wound,and improve the quality of life of patients.In addition,no adverse reactions related to Kangfuxin solution were found during the investigation.2.The results of network pharmacology and molecular docking showed that Cyclo（L-Val-L-Tyr）,Cyclo（L-Pro-L-Tyr）and Cyclo（L-Tyr-L-Tyr）in Kangfuxin solution might be the core active ingredients in the treatment of oral ulcer.Through key targets such as STAT3,MAPK1,AKT1 and MMP9,it plays an important role in the PI3K-Akt signaling pathway,RAP1 signaling pathway and TNF signaling pathway.The results of molecular docking between the core compounds and the targets showed that there was a good affinity between the ligand and the receptor,and the binding of Cyclo（L-Tyr-L-Tyr）to AKT1 and MMP9 was the most stable.In addition,the active flavonoids in Kangfuxin Liquid: 7-hydroxyflavone,flavanone and 7,4’-dihydroxyflavone may play a key role in promoting wound repair through acting on EGFR,ADORA2 a,DRD2,HTR2 A,CHRNA7 and other proteins in the signal pathway of neuroactive ligand receptor interaction,calcium signal pathway and c AMP signal pathway.3.The results of cell proliferation and scratch experiments showed that 7-hydroxyflavone,flavanone and 7,4’-dihydroxyflavone could significantly promote the proliferation of CCD841 when the concentration was 10 μM/ml and the time was 24 h.At 12 h and 24 h,7-hydroxyflavone and flavanone significantly promoted the migration of CCD841.In addition,7-hydroxyflavone and 7,4’-dihydroxyflavone significantly upregulated the m RNA expression of EGFR.Conclusions:1.As a traditional Chinese medicine preparation that has been on the market for more than 30 years,kangfuxin liquid has a relatively rich basis of clinical application,and has shown good safety and effectiveness in clinical use.At the same time,pharmacoepidemiological investigation provides scientific reference for re-evaluation of drugs after marketing,discovery of early pharmacovigilance signals and indications for new drugs,and rational clinical use of drugs,which has certain prospective significance.2.The application of network pharmacology and molecular docking technology fully reveals the theoretical system of "multi-component,multi-target and multi-channel synergy" of traditional Chinese medicine,which provides a new insight for evaluating the clinical efficacy and safety of kangfuxin liquid in the treatment of ulcer/wound,as well as the study of active components and molecular mechanism.At the same time,it is of great significance to the design of new drugs and the improvement of the API and preparation of the marketed drugs.3.Flavonoids,as active components widely existing in traditional Chinese medicine,have pharmacological effects such as anti-inflammatory,anti-oxidation and anti-gastric ulcer.This study explored its effects on the proliferation and migration of CCD841 cells,so as to provide reference for the follow-up study of flavonoids metabolism in vivo and in vitro. |
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