Objective: To study the mechanism of Salviae Miltiorrhizae Radix (SMR) et Rhizoma and Carthamii Flos (CF) in the treatment of myocardial infarction (MI) by means of network pharmacology and experimental verification. Methods: The main components of SMR-CF were searched by Traditional Chinese Medicine System Pharmacological Analysis Platform (TCMSP), and the component targets were screened by TCMSP and Swiss Target prediction databases, and the MI related targets were queried through OMIM, TTD, Genecards and NCBI (Gene) databases. The common target proteins of disease and drug components were screened by the intersection of drug targets and disease targets. The network model of protein-protein interaction (PPI) was constructed by using STRING platform. The functional enrichment analysis of gene ontology (GO) and the KEGG pathway were carried out by using DAVID. Cytoscape was used to construct medicinal material-target and medicinal material-composition-target-pathway network map for further experimental verification, in order to reveal the therapeutic effect of SMR-CF on MI in mice. Results: A total of 84 active components were screened from SMR and CF, 485 target proteins were searched, and 28 targets related to MI were found. GO functional enrichment analysis showed that there were 18 GO entries, including 9 biological process (BP) entries, three molecular functional (MF) entries and six cell composition (CC) entries. KEGG pathway was enriched and screened to obtain 30 signal pathways, such as hypoxia inducible factor signaling pathway, tumor necrosis factor, vascular endothelial growth factor, sheath phospholipid, small G protein Rap1 and so on. The results of HE staining and Masson staining showed that SMR-CF could significantly improve myocardial injury. Western blotting results showed that SMR-CF could down-regulate the expression of TNF-α and MAPK to improve myocardial injury. Conclusion: The target and pathway of SMR-CF in the treatment of MI are explained by network pharmacology, which provides a scientific basis for the further clarification of SMR-CF in the treatment of MI.

In order to study the important factors involved in cationic liposome-mediated gene transfer, Lipofectamine 2000 or DOTAP was evaluated using three types of cells (Hep-2, MCF-7 and SW-480) in vitro transfection efficiencies. Different properties of the two reagents were analyzed and compared by DNA arrearage assay and MTT assay. Both Lipofectamine 2000 and DOTAP had strong capability to combine with DNA; Lipofectamine 2000 can get higher transfection efficiency of the three cells by using GFP as report gene, meanwhile, DOTAP can also get higher transfection efficiency against Hep-2 cell. However, DOTAP showed lower transfection efficiency against MCF-7 and SW-480 cell. On the other hand, the cytotoxicity assay showed that over 85% cell viability of MCF-7 cell could be achieved both by Lipofectamine 2000 and DOTAP under the optimal transfection condition. Relatively speaking, Lipofectamine 2000 has very high transfection efficiency in a broad range of cell lines, but because of the special selectivity of cell type on liposome, DOTAP also has a broad application prospect.
Cell Line, Tumor ; Cell Survival ; drug effects ; DNA ; genetics ; Fatty Acids, Monounsaturated ; chemistry ; toxicity ; Gene Transfer Techniques ; Genes, Reporter ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Lipids ; chemistry ; toxicity ; Quaternary Ammonium Compounds ; chemistry ; toxicity ; Transfection

Animals ; Bacterial Vaccines ; Brucella suis ; genetics ; Brucellosis ; epidemiology ; microbiology ; veterinary ; China ; epidemiology ; DNA Fingerprinting ; DNA, Bacterial ; genetics ; Electrophoresis, Gel, Pulsed-Field ; Humans ; Nucleic Acid Amplification Techniques ; methods ; Time Factors

Objective: To investigate the protective effect of compound Longmaining isoprenaline hydrochloride-induced myocardial infarction model and its effect on Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear transcription factor-kappa B (NF-κB) signaling pathway.Method: Forty-eight male SD rats were randomly divided into 6 groups:normal group,model group,compound salvia miltiorrhiza drop pill group (0.072 9 g·kg^-1),and low,medium and high-dose compound Longmaining decoction groups (0.36,0.71,1.43 g·kg^-1).The acute myocardial infarction model was induced through subcutaneous injection with isoproterenol.The pathological changes of myocardial tissue were examined by hematoxylin-eosin (HE) staining.The levels of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),monocyte chemotaxis protein-1(MCP-1) and nitrogen (NO) in serum were measured by enzyme linked immunosorbent assay (ELISA).The expression levels of inhibitors of NF-κB kinase subunit-β(IKKβ),NF-κB inhibitor α(IκBα),TLR4,MyD88 and NF-κB p65 were measured by immunohistochemical staining and Western blot.Result: Compared with normal group,the myocardial injury in model group was obvious.The levels of IL-1β,IL-6,TNF-α,MCP-1 and NO in serum increased significantly (P<0.05),the expression level of IκBα decreased significantly (P<0.05),and the expression levels of IKKβ,TLR4,MyD88 and NF-κB p65 increased significantly in myocardial tissue (P<0.05).Compared with model group,low,medium and high-dose compound Longmaining groups could significantly alleviate the degree of myocardial injury in rats,significantly decrease IL-1β,IL-6,TNF-α,MCP-1 and NO levels in the serum (P<0.05),significantly increase the expression level of IκBα(P<0.05),and decreased the expression levels of IKKβ,TLR4,MyD88 and NF-κB p65(P<0.05).Conclusion: Compound Longmaining plays a protective effect on acute myocardial infarction by regulatingthe expressions of TLR4/MyD88/NF-κB p65 signaling pathway and relevant inflammatory factors.