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OBJECTIVE To explore whether diterpenoid 12-deoxyphorbol-13-palmitate （DP） from Euphorbia fischeriana can exert anti-leukemia effects through the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signal pathway， and to provide experimental evidence for developing it into a new anti-leukemia drug. METHODS Using LY294002 （PI3K specific inhibitor） as tool drug， the effects of 24 h DP treatment on the proliferation and apoptosis of human myeloid leukemia HL60 cells were detected by MTT method， Annexin Ⅴ-FITC/PI staining and AO-EB staining. ELISA method was used to detect lactic dehydrogenase （LDH） release and the activities of cysteinyl aspartate specific proteinase 3 （caspase-3） and caspase-9. The transcriptional level of caspase-3， caspase-9， forkhead box O3a （FoxO3a） and B cell lymphoma 2 interacting mediator of cell death （Bim） mRNA were detected by real-time quantitative polymerase chain reaction （qRT-PCR）. The protein expression of phosphorylated FoxO3a （p- FoxO3a） and phosphorylated Akt （p-Akt） were detected by Western blot method. The nuclear translocation of FoxO3a protein was detected by immunostaining combined with laser confocal microscopy. RESULTS 10 μmol/L DP and 10 μmol/L DP+LY294002 could inhibit the proliferation and induce the apoptosis of HL60 cells （P＜0.01）. After treatment of 5， 10， 20 μmol/L DP， HL60 cells showed typical morphological characteristics of apoptosis； DP could significantly increase the levels of LDH release and the activities of caspase-3 and caspase-9 （P＜0.05 or P＜0.01）， in dose-dependent manner. After treatment of 10 μmol/L DP and 10 μmol/L DP+LY294002， the transcriptional levels of caspase-3， caspase-9 and Bim mRNA were increased significantly （P＜0.05 or P＜0.01）， and transcriptional level of FoxO3a mRNA and protein expressions of p-FoxO3a and p-Akt were decreased significantly （P＜0.05 or P＜0.01）. Nuclear translocation changes were observed in FoxO3a protein in 10 μmol/L DP+LY294002 group， and the change was more significant than that of LY294002 group. CONCLUSIONS DP can inhibit the proliferation and induce the apoptosis of HL60 cells via inhibiting PI3K/Akt signaling pathway.
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Objective:To virtual screen the active ingredient of Shenmai injection in treatment of Novel Coronavirus Pneumonia and discuss the potential mechanism based on network pharmacology and high throughput molecular docking. Methods:Based on network pharmacology and high-throughput molecular docking technology, the compounds and predicted targets of Shenmai injection were retrieved from TCMSP, BATMAN-TCM and Targetnet databases, and the composition target map was constructed. The genes related to coronavirus pneumonia were retrieved from OMIM and GeneCards databases, and the PPI network between target genes was constructed by searching the common parts of target genes; David 6.8 was used to analyze gene function and pathway enrichment, and PDB database was used to obtain protein crystal structure, and Autodock Vina and python scripts were used for high-throughput molecular docking. Results:A total of 27 compounds and 224 target genes were obtained. 15 core components and 15 core targets for the treatment of coronavirus pneumonia were identified: CASP3, NOS2, PARP1, CASP8, NOS3, BCL2, ADA, OPRM1, TGFB1, TLR9, ACHE, SLC29A1, BAX, ADK, and PNP. The enrichment analysis showed that the core targets acted on the signaling pathways such as Tuberculosis, Pathways in cancer, Hepatitis B and Apoptosis. The better components of Novel Coronavirus Pneumonia related targets were diosgenin, stigmasterol, beta-sitosterol and ginsenoside Rh1_qt obtained by virtual screening.Conclusion:This study screened out the active ingredient and tarket of Shenmai Injection in treatment of Novel Coronavirus Pneumonia. It laid a foundation for the further clinical application of Shenmai injection and development of novel coronavirus pneumonia drugs.
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Objective:With network pharmacology, this paper aims to explore the potential active constituents, related targets and signal pathways to elucidate the mechanism of Calyx seu Fructus Physalis in the treatment of lung cancer. Methods:By searching for Pharmacology Database of Traditional Chinese Medicine Systems and Analysis Platform (TCMSP) to obtain the active constituents of Calyx seu Fructus Physalis, and predict the target genes of active constituents by using SWISS and SEA. By collecting the corresponding targets approved by FDA and search for different database, including Therapeutic Target Database (TTD), Database of Gene-disease Associations (DisGeNET) and Online Mendelian Inheritance in OMIM to build a database of lung cancer related target genes; To get the target genes of herb-disease proteins by the intersection of the two databases and display the results through the network software of Cytoscape 3.7.2. Then to use the Database of Annotation, Visualization and Integrated Discovery (DAVID) to conduct key enrichment analysis of Kyoto Gene and Genome Encyclopedia (KEGG) pathway enrichment analysis. Combining related literature to analize the active ingredients and mechanism of Calyx seu Fructus Physalis in the prevention and treatment of lung cancer. Results:It was found that 11 active constituents of Calyx seu Fructus Physalis could play the role of anti-lung cancer by regulating 19 lung cancer related targets such as EGFR, ESR1, MDM2, MMP2 and MET. There were 15 signal pathways involved Proteoglycans in cancer, MicroRNAs in cancer, Pathways in cancer, Transcriptional misregulation in cancer, PI3K-Akt signaling pathway and other key signal pathways. Conclusion:The results show that the active constituents of anti-lung cancer effect of Calyx seu Fructus Physalis may be Oleic acid, Cycloartenol, Obturator, Graminesterol, β-sitosterol, etc. The mechanism of action may be related to multiple cancer-related signaling pathways such as Proteoglycans pathway, Transcriptional misregulation pathway, and PI3K-Akt signaling pathway.
