In Vitro And In Vivo Material Foundation Of Qihuang Jianpi Zishen Granules And Network Pharmacology Study On Systemic Lupus Erythematosus

Objective:Qihuang Jianpi Zishen Granules(QHJPZSG)is an in-hospital preparation for the treatment of systemic lupus erythematosus(SLE)in the First Affiliated Hospital of Anhui University of Chinese Medicine.Currently,the related research mostly focuses on the clinical,and there is no basic research on the in vivo and in vitro substances of QHJPZSG,which hinders the clinical application.the changes of target organs in mice after taking medicine were observed,and ultra-high performance liquid coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS~E)was used to explore the material basis of QHJPZSG in vitro and in vivo,and combined with network pharmacology to predict the main components,targets and related pathways of QHJPZSG in the treatment of SLE,so as to provide a basis for the subsequent related research of QHJPZSG.Methods:The kidneys of mice in each group were observed by HE staining.UPLC-Q-TOF-MS~E was used to collect the test solution and mixed reference substance solution of QHJPZSG,and the results were imported into UNIFI software together with the self-built database for matching.Masslyxn4.1 software was used to check the results of UNIFI software with the reference substance fragment information and relevant literature,and the mass spectrometric cleavage rules of various chemical components were summarized to clarify their components and attribution.The rats were intragastrically administered with QHJPZSG and blood was taken from the canthus.After the serum samples were prepared,the serum containing the drugs was analyzed by UPLC-Q-TOF-MS~E to clarify the prototype components and metabolites of QHJPZSG in vivo.The Swiss Target Prediction database and the GEO database are used for respectively predicting prototype components in vivo and a target of SLE,after an intersection target is taken,the key components are screened out by using Cytoscape,a protein interaction network is constructed by using a STRING database,a result is visualized and the key target is screened out by using Cytoscape,and the intersection target is introduced into a DAVID database,The enrichment of GO function in the database and the annotation function of KEGG pathway were used to identify the gene functions and related pathways involved,and the molecular docking of key components and targets was performed using Auto Dock Vina software.Results:Compared with the normal group,the kidneys of mice in the model group were damaged seriously and the inflammatory cells were increased significantly.Compared with the model group,the kidneys of mice in the administration group significantly alleviated the above symptoms.A total of 114 compounds were identified by UPLC-Q-TOF-MS~E in combination with UNIFI software and manual collation by Masslyxn4.1 software.Among them,18 were derived from Radix Astragali seu Hedysari,27 from Radix Rehmanniae Preparata,19 from Semen Cuscutae,9 from Rhizoma Dioscoreae,5 from Rhizoma Atractylodis Macrocephalae,14 from Fructus Rosae Laevigatae,17 from Fructus Rubi,18 from Poria,and 11 were common compounds of multiple medicinal materials.The main components of QHJPZSG were flavonoids,terpenes,organic acids,phenylethanoid glycosides,saponins and lignans.Using 114 chemical components of QHJPZSG as the database of prototype components in vivo,a total of 27 prototype components and 51 metabolites were identified through UPLC-Q-TOF-MS~E combined with UNIFI software and Masslyxn4.1 software manual verification,with flavonoids and terpenes as the main prototype components,and hydrolysis,decarboxylation and methylation as the main metabolic pathways.By the network pharmacology method,521 component targets and 3337 disease targets were predicted in the Swiss Target Prediction database and GEO database,respectively,with 141 intersecting targets.There were 10 key components,19 key targets,and 46 pathways involved in the treatment of SLE with QHJPZSG.The results of molecular docking indicated that the key components and key targets had good binding capacity.Conclusion:QHJPZSG can alleviate the kidney damage caused by SLE.The in vitro and in vivo material bases of QHJPZSG were identified using UPLC-Q-TOF-MS~E and UNIFI software.Based on the research method of network pharmacology,it was shown that in the treatment of SLE,QHJPZSG might act on such targets as EGFR,ESR1,MMP9,CCND1,and MDM2 as well as the neuroactive ligand-receptor interaction,Calcium signaling pathway,the regulation of inflammatory mediators of cancer polysaccharide protein pathway and TRP channel,through such components as Diosmetin,Biatractylolide,Isorhamnetin,Atractylenolide III,and Formononetin.