Therapeutic Effect Of Saposhnikovia Divaricata On Type Ⅰ Allergy

Saposhnikovia divaricata,as a widely used Chinese medicine,is often used to treat headache,dizziness,fever,rheumatoid arthritis and other diseases.Recent studies have shown that Saposhnikovia divaricata has various pharmacological effects such as anti-inflammatory,immune regulation,anti-tumor,and potential anti-allergic effect.Howener,the detailed analysis of Saposhnikovia divaricata in type I allergy has not been reported.In this study,the molecular mechanism of Saposhnikovia divaricata on type I allergy was exlpored preliminarily using network pharmacology.And moreover,the therapeutic effect and molecular mechanism of Saposhnikovia divaricata on type I allergy were evaluated by RBL-2H3 cell and passive skin allergy mouse in vitro and vivo,respectively.The main research contents are as follows:1.Network pharmacological analysis: The potential effective components and action targets of Saposhnikovia divaricata were found through TCMSP database,and then the target genes related to "allergy" were found through Gene Cards and OMIM database.The obtained target intersections were imported into the David database for GO enrichment analysis.The intersection of the targets obtained in the previous two steps were processed by Bioconductor package called by the R software to get KEGG enrichment analysis.The results showed that Saposhnikovia divaricata may play its role in treating allergic diseases through multi-component and multi-target regulation of calcium signal,PI3K/Akt and MAPK.2.In vitro experiment: CCK-8 method was used to detect the adverse effect of Saposhnikovia divaricata and Prim-O-glucosylcimifugin(an index component of Saposhnikovia divaricata)on RBL-2H3 cells to screen the optimum concentration of Saposhnikovia divaricata and Prim-O-glucosylcimifugin on RBL-2H3 cells.The effects of both on the release of β-Hex and histamine in RBL-2H3 cells were detected by chromogenic method and ELISA.The apoptosis of cells and its effect on the concentration of calcium ions in cells were observed by AO/EB method,The expression of key genes in signal pathway related to type I allergy during degranulation was detected by fluorescence real-time quantitative PCR and Western blot.The results showed that Saposhnikovia divaricata had no obvious toxic effect on RBL-2H3 cells when the crude drug concentration of Saposhnikovia divaricata was 0～4 mg/m L or Prim-O-glucosylcimifugin was at the concentration of 0～100μg/m L.Within the safe concentration range of Saposhnikovia divaricata,both Saposhnikovia divaricata and Prim-O-glucosylcimifugin can inhibit the release of β-Hex and histamine induced by DNP-Ig E/DNP-BSA in a concentration-dependent manner,but have no inhibitory effect on the release of β-Hex and histamine induced by C48/80 cells.In addition,Saposhnikovia divaricata can obviously inhibit the release of intracellular calcium and apoptosis induced by DNP-Ig E/DNP-BSA,but has no inhibitory effect on the release of calcium induced by C48/80.The results of real-time quantitative PCR and Western Blot showed that Saposhnikovia divaricata may play a therapeutic role on type I allergy by inhibiting Lyn/Syk,PI3K/AKT/NF-κB and MAPK signal transduction pathways.3.In vivo experiment: Saposhnikovia divaricata and Prim-O-glucosylcimifugin were continuously administered to mice by gavage for one week,and the PCA model of mice was established by DNP-Ig E/DNP-BSA and C48/80 induction.The therapeutic effect of Saposhnikovia divaricata on type I allergy was evaluated by detecting Evans blue exudation and swelling degree in ears and toes of mice in PCA model.The result showed that both Saposhnikovia divaricata and Prim-O-glucosylcimifugin could inhibit Evans Blue exudation and tissue swelling in mouse ear and toe PCA model induced by DNP-Ig E/DNP-BSA,which confirmed that Saposhnikovia divaricata had significant therapeutic effect on type I allergy at the animal level as a whole,but they had no significant therapeutic effect on mouse PCA model induced by C48/80.To sum up,through network pharmacological analysis,RBL-2H3 cell test and passive skin allergy test in mice,this paper proves that Saposhnikovia divaricata has a significant therapeutic effect on type I allergy,which may regulate the pathogenesis of allergy through Lyn/Syk/PLCγ,PI3K/AKT/NF-κB,MAPK signal transduction pathways,and plays a therapeutic role on type I allergy.