基于网络药理学和动物实验探讨升陷汤促进膀胱逼尿肌细胞增殖的作用机制OA

[Objective]To investigate the therapeutic effect and mechanism of Shengxian Decoction on bladder detrusor injury.[Methods]Based on the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),the active components and drug targets of Shengshen Decoction were screened;the related targets of urinary retention were obtained from GeneCards,Therapeutic Target Database(TTD)and DrugBank database.The"traditional Chinese medicine-active component-target"network and protein-protein interaction(PPI)network were constructed to screen the core targets.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were performed to construct a"pathway-target-component"network.Molecular docking was employed to verify the binding ability of core components to their targets.Through bladder neck and urethra ligation,a rat model of partial bladder outlet obstruction(pBOO)was established.After modeling,the rats were randomly divided into sham operation group(Sham),model group(Mol),and Shengxian Decoction low(SXTL,3.9 g·kg-1),medium(SXTM,7.8 g·kg-1),high dose group(SXTH,15.6 g·kg-1),western medicine group[Tamsulosin-Pyridostigmine Bromide(TS-PB),Pyridosine Bromide(18 mg·kg-1)combined with Tamsulosin capsule contents(0.02 mg·kg-1)].After 4 weeks of intervention,the bladder mass index,histomorphology,cell proliferation,mRNA and protein expression of cyclic adenosine monophosphate(cAMP),protein kinase A(PKA),and cAMP-response element binding protein(CREB)were detected.[Results]A total of 1 117 active ingredients of Shengxian Decoction,233 targets of action and 210 intersection targets of urinary retention were selected.The core targets included tumor protein 53(TP53),protein kinase B(AKT1)and Jun kinase(JNK).KEGG enrichment analysis showed that target genes were mainly enriched in the following signaling pathways:phosphoinositide 3-kinase/protein kinase B signaling pathway(PI3K/AKT),mitogen-activated protein kinases(MAPK),hypoxia inducible factor-1(HIF-1)and cAMP signaling pathways.The core active ingredients were quercetin,kaempferol,luteolin,7-O-methylisomucinol,and trichomycin.Shengxian Decoction treatment significantly reduced the bladder weight/body weight ratio(P<0.001,P<0.0001),reduced collagen fiber deposition(P<0.0001),down-regulated the mRNA expression of cAMP,PKA and CREB(P<0.01,P<0.001,P<0.0001),and inhibited the protion expression of cAMP,p-PKA/PKA and p-CREB/CREB(P<0.05,P<0.01,P<0.0001),SXTH showed particularty good effect(P<0.0001).[Conclusion]Shengxian Decoction can inhibit the cAMP signaling pathway,alleviate bladder fibrosis and promote the proliferation of bladder detrusor muscle cells through the synergistic action of multi-components,multi-targets and multi-pathways,so as to improve bladder structure and function after pBOO.