Action Mechanism Of Shenghua Decoction In Treating Postpartum Lochia Based On Network Pharmacology

ObjectiveTo confirm the main active components and targets of Shenghua Decoction(SHD)by the method of network pharmacology,to explore the mechanism of its multi-component,multi-target and multi-pathway action in the treatment of postpartum lochia(PL),and to select the PI3K/AKT/NF-κB signaling pathway for animal experimental validation.MethodsSHD monomer components were searched by TCMSP,TCMID and TCM@taiwan databases,and SHD active compounds and targets were screened with "OB≥30%,DL≥0.18" to construct SDH active compound-target network maps for topological parameter analysis.The targets of SHD in the treatment of PL were also screened according to the targets associated with PL in the TTD and OMIM databases,and the mechanism of SHD in the treatment of PL was explored by gene ontology(GO)functional enrichment analysis and KEGG pathway analysis of the targets.Animal experiments were used for verification: 60 SD rats on the first day after delivery were divided into 5 groups according to the random number table: blank group(Control),model group(Model),Shenghua Decoction group(Model+S),inhibitor group(Model+LY),and Shenghua Decoction+inhibitor group(Model+LY+S),with 12 rats in each group.Among them,except that the blank group was not inoculated with pathogens in utero,the other 4 groups were inoculated with pathogens in utero,and then the PL model was established.On the 4th day after delivery,the control group was given 5 ml/kg triple distilled water by gavage,along with 10 mg/kg normal saline by intraperitoneal injection.The Model group was the same as the control group.The Model+S group was given Shenghua Decoction 5 ml/kg by gavage,along with 10 mg/kg normal saline by intraperitoneal injection.The Model+LY group was given 5 ml/kg triple distilled water by gavage,along with PI3K/AKT inhibitor LY294002 10 mg/kg by intraperitoneal injection.The Model+LY+S group was given 5 ml/kg Shenghua Decoction by gavage along with LY294002 10 mg/kg by intraperitoneal injection,once daily at 8 am for 7 consecutive days in all groups.ELISA was used to detect the serum tumor necrosis factor alpha(TNF-α)level in rats.HE staining was used to analyze uterine morphology.Immunofluorescence and Western blot were used to detect the expression of PI3K/AKT/NF-κB signaling pathway protein in rat uterine tissues.Results1.In this study,120 active components of SHD,236 predicted targets and 1053 PL related targets were obtained,and 84 common targets of SHD and PL were selected.The targets with larger degree were TP53(61),JUN(60),AKT1(60),FSR1(53),PTEN(49)and MTOR(49).2.The PI3K/AKT/NF-κB signaling pathway was verified by animal experiments.The results of gross uterine morphology and HE staining showed that compared with the control group,the uterus of rats in the model group was enlarged,the weight was increased,the uterine wall was thickened,and pus like fluid was observed in the uterine cavity.Endometrial continuity was interrupted,epithelial degeneration,necrosis,interstitial hemorrhage,edema,and massive inflammatory cell infiltration were observed.After SHD treatment,there was occasional interruption of continuity in the endometrium,less epithelial damage,and fewer inflammatory cells in the Model+S group.The degree of uterine injury in the Model+LY group and Model+LY+S group was milder than that in the Model+S group,and heavier than that in the Model+S group.ELISA results showed that the level of serum TNF-α in the Model group was significantly increased compared with that in the Control group(P<0.01).After SHD treatment,the level of serum TNF-α in the Model+S group was significantly decreased compared with that in the Model group(P<0.01).Immunofluorescence results showed that the expression of p-PI3 K and p-AKT in the Model+S group was decreased compared with that in the Control group,while after SHD treatment,the expression of p-PI3 K and p-AKT in the Model+S group was increased compared with that in the Model+S group,and there was no significant difference in the expression of p-AKT between the groups.The expression of p-NF-κB p65 protein in the Model+S group and Control group was significantly lower than that in the Model group(P<0.01),and the expression of p-NF-κB p65 protein in the Model+LY group and Model+LY+S group was lower than that in the Model group(0.05<P<0.01).SHD significantly increased PI3 K,AKT,and NF-κB p65 protein phosphorylation levels(P<0.01)and significantly decreased serum TNF-α expression levels(P<0.01)in the PL rat model.ConclusionsThe 84 targets obtained by network pharmacology may be potential targets for the treatment of PL by SHD,and their mechanisms of action may be related to signaling pathways such as PI3K/AKT,MAPK,HIF-1 and FXO.The selected PI3K/AKT/NF-κB signaling pathway was verified by animal experiments,indicating that this pathway is a potential molecular mechanism of SHD in the treatment of PL,which lays a research foundation for deeply exploring the mechanism of SHD in the treatment of PL through the way of multi-component,multi-target and multi-pathway.