Systematic elucidation of the traditional Chinese medicine prescription Danxiong particles via network pharmacology and molecular docking

Purpose: To investigate the pharmacological effect of the traditional Chinese medicine (TCM) prescription Danxiong particles (TDX105) and its mechanism of action.Methods: The active compound and targets of TDX105 were investigated via network pharmacology. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were enriched, and protein-protein interaction network (PPI) was constructed. A network of ‘components-targets-pathways’ was developed with Cytoscape 3.8.0 software, while the formation of molecular docking analysis was conducted using Autodock vina software.Results: There were 304 compounds and 482 targets identified in total. Genes with degree ≥ mean node values were selected as the crucial targets, and string database was to be combined to 64 targets identified with cytoscape so as to draw a protein interaction map. A total of 137 pathways were enriched from 64 targets involving mainly 10 pathways, for example, PI3K-Akt signaling pathway, pathways in cancer, human cytomegalovirus infection and focal adhesion. Then, compound-target and compoundtarget- pathways were constructed using cytoscape (3.8.0). Finally, the five most active compounds, viz, quercetin, myricetin, luteolin, ellagic acid and kaempferol, and the top ten targets AKT1, GAPDH, TP53, ALB, EGFR, MAPK3, JUN, MAPK1, SRC and ESR1 were selected for molecular docking. These targets and compounds had strong interactions through a combination of hydrogen bonds and hydrophobic forces.Conclusion: The mechanism of action of TDX105 has been successfully explained using the combination of network pharmacology and molecular docking. This may offer a solid foundation to the clinical use of TDX105, and further strengthen the prospects of its development for clinical use

Molecular Cloning and functional characterization of a putative Elovl4 gene and its expression in response to dietary fatty acid profiles in orange-spotted grouper Epinephelus coioides

Elongase of very long-chain fatty acids (Elovl) 4 probably plays a crucial role in marine fish species, where lack of Elovl2 has been considered as one possible reason for their low long-chain polyunsaturated fatty acids' (LC-PUFAs) biosynthetic capability. Elongase of very long-chain fatty acids 4 is the most recent member of the Elovl family that has been investigated in fish. Here, we report the molecular cloning and functional characterization of putativeelovl4cDNA isolated from marine teleost,Epinephelus coioides, and its expression in response to dietary n-3 LC-PUFA and docosahexaenoic acid (DHA) to eicosapentaenoic acid (EPA) ratio. Theelovl4cDNA of grouper was 2341bp including 301bp of 5′-untranslated region (UTR), 918bp of the coding region that encodes 305 amino acids (AA) and 1122bp of 3′UTR. Heterologous expression in yeast demonstrated that grouper Elovl4 could elongate saturated fatty acids (FA), especially 24:0 and 26:0, up to 36:0. Also, grouper Elovl4 effectively converted C20 and C22 polyunsaturated FAs to elongated polyenoic products up to C36. Tissue distribution analysis revealed that Elovl4 were widely transcribed in various tissues with the highest level in eye, brain and testis as described in other teleosts. The transcript level ofelovl4was significantly affected by dietary n-3 LC-PUFA and high LC-PUFA level repressess its expression. However, the ratio of DHA to EPA had no significant influence on its expression. These results may contribute to better understanding the LC-PUFA biosynthetic pathway in this fish species