ABSTRACT
Objective:To explore the active components， targets， and signaling pathways responsible for Bushen Zhuyun prescription in treating the recurrent spontaneous abortion （RSA） based on network pharmacology and uncover its potential mechanism by molecular docking and in vitro cell experiments. Method:The active components of Bushen Zhuyun prescription were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， Traditional Chinese Medicine Integrated Database （TCMID） and the published articles， followed by the prediction of drug action targets based on such platforms as DrugBank and SwissTargetPrediction. GeneCards and Online Mendelian Inheritance in Man （OMIM） were searched to obtain the RSA targets， which were then intersected with the targets of Bushen Zhuyun Decoction. Following the plotting of Bushen Zhuyun prescription-compound-target-RSA network by Cytoscape 3.7.1， the protein-protein interaction （PPI） network was then constructed with STRING for screening the core network. The resulting common targets were then subjected to Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis using R software. Autodock Vina 1.1.2 was used for molecular docking. The activation of phosphatidylinositol 3-kinase/protein kinase B （PI3K/AKT） signaling pathway by Bushen Zhuyun prescription was verified in human umbilical vein endothelial cells （HUVEC） <italic>in vitro</italic>. Result:It was found that 49 potential active components of Bushen Zhuyun prescription might act on 133 RSA targets. GO enrichment analysis yielded 470 biological processes， with angiogenesis， vascular development， cellular proliferation， and oxidative activity mainly involved. KEGG enrichment analysis revealed 103 signaling pathways （<italic>P</italic><0.05）， and the PI3K/AKT signaling pathway， advanced glycation end product （AGE）/receptor for advanced glycation end product （RAGE） signaling pathway， and tumor necrosis factor （TNF） signaling pathway were the main ones. As indicated by molecular docking， the Vina scores of the main active component kaempferol with AKT1 and vascular endothelial growth factor A （VEGFA） were the lowest and similar. It was confirmed <italic>in vitro</italic> cell experiments that Bushen Zhuyun prescription activated the PI3K/AKT signaling pathway and up-regulated the expression of VEGFA and downstream AKT protein to promote angiogenesis. Conclusion:Bushen Zhuyun prescription promotes angiogenesis at the maternal-fetal interface by regulating angiogenesis and cellular proliferation， activating the PI3K/AKT pathway， and up-regulated the VEGFA expression， which is beneficial to the formation of placenta in early pregnancy and the maintenance of early pregnancy. This study has provided ideas for new drug development.
ABSTRACT
<p><b>OBJECTIVE</b>To explore the possibility of cell-medicated immune response induced with heat shock protein 70 (HSP70)-HBsAg protein complex in vitro.</p><p><b>METHODS</b>HSP70-HBsAg complex was reconstituted in vitro which was injected into mice in order to observe that whether HSP70-HBsAg would stimulate humoral and cellular immune responses. HSP70, HSP70-HBsAg complex and HBsAg were used to activate the dentritic cell (DC) individually, which would initiate homogeneic T lymphocyte to transform to cytotoxic T lymphocyte (CTL). The cytotoxicity of CTL was detected with MTT assay.</p><p><b>RESULTS</b>HSP70-HBsAg complex elicited both humoral and cellular immune responses against HBsAg in mice. Specific CD8+ CTL response was readily induced by HSP70-HBsAg complex and HBsAg, especially the former.</p><p><b>CONCLUSIONS</b>HSP70-HBsAg complex is immunogenic and HSP70 can lead to great efficient CTL response. And HSP70-HBsAg complex may be used as a protein vaccine for immunotherapy for chronic hepatitis B.</p>