ABSTRACT
Aim To clarify the mechanism of Gonglaoye and Xianhecao herbal pair in the treatment of ischemic stroke so as to obtain the substantive evidence using network pharmacology data mining and molecular docking. Methods The main compounds of traditional Chinese medicine were obtained by TCMSP platform and consulting literature, the drug action targets were obtained by TCMSP, and the known genes about ischemic stroke were collected by searching Drugbank, Disgenet, TTD, Genecards, OMIM database, thus the drug-compound-target network map was constructed, and the common target proteins and main compounds were screened. The visual protein-protein interaction network map (PPI) was constructed by string. With the help of Cytoscape software, the original target network of active components was constructed and analyzed, and the gene ontology GO and Jingdu gene and genome encyclopedia KEGG analysis were carried out to analyze the GO function and KEGG pathway enrichment of the common targets of drugs and diseases. Finally, the molecular docking of the core protein and the core compound was carried out according to the relevant node parameters of the compound and protein. Results Seventeen active components and 296 potential targets of Gonglao leaf and crane herbs in the treatment of ischemic stroke were screened. GO enrichment was mainly concentrated in the response to oxides, cell response to chemical stimulation, positive regulation of cell metabolism, constant effect, active regulation of stimulus response, cell communication and so on. KEGG was mainly involved in signaling pathways such as PI3K-Akt, Ras, neuron ligand receptor interaction and so on. Molecular docking showed that quercetin and other active components had high affinity and tight connection with core targets such as AKT1. Conclusions The treatment of ischemic strokec is mainly through the mechanism of ursolic acid, hyperin and other active components, AKT1, cMAPK3 and other multi-targets, PI3K-AKT and other multi-pathway interaction mechanisms. Through this study the theoretical support can be provided for the further clinical application of Gonglaye and crane herbs, providing basic ideas for future experimental research and new drug research and development.
ABSTRACT
Objective · To investigate the effect of a new type of allene compound, 1-phenylpropadienyl phosphine oxide (PHPO), on proliferation and apoptosis of lung cancer cell line A549. Methods · A549 cells were treated with different concentrations of PHPO. The effects of PHPO on cell proliferation, apoptosis and cell cycle were detected by CCK-8 and flow cytometry assay. Wound healing test was used to measure the migration ability of A549 cells. Real-time PCR was used to detect the expression of apoptosis and cell cycle related gene. The expression of proteins in MAPK pathway was assayed by the Western blotting. The nude mice xenograft model of human lung cancer A549 cells was established. After tumor formation, PHPO was injected daily for treatment, and the tumor size was observed. Results · Compared to the control group, PHPO significantly inhibited the cell viability of A549 cells and induced apoptosis of them, and the IC50 value of 24 h is 44.23 μmol/L. PHPO blocked the cell cycle in the G1 phase significantly. The migration capacity of PHPO-treated cells was decreased. The mRNA levels of Bax and P21 were up-regulated in PHPO-treated group, and the mRNA lever of Bcl-2 was down-regulated (P<0.05). PHPO increased the phosphorylation levels of p38, ERK and JNK. Injection of PHPO could significantly inhibit the growth of tumor in the xenograft model compared to the control group (P<0.05). Conclusion · PHPO can induce the apoptosis and inhibit the proliferation of A549 cells, block the cell cycle in the G1 phase and decrease the migration ability of A549 cells significantly. The mechanism may be related to the activation of MAPK signaling pathway by PHPO and the increase of phosphorylation of p38, ERK and JNK.
ABSTRACT
Objective · To investigate the effect of a new type of allene compound, 1-phenylpropadienyl phosphine oxide (PHPO), on proliferation and apoptosis of lung cancer cell line A549. Methods · A549 cells were treated with different concentrations of PHPO. The effects of PHPO on cell proliferation, apoptosis and cell cycle were detected by CCK-8 and flow cytometry assay. Wound healing test was used to measure the migration ability of A549 cells. Real-time PCR was used to detect the expression of apoptosis and cell cycle related gene. The expression of proteins in MAPK pathway was assayed by the Western blotting. The nude mice xenograft model of human lung cancer A549 cells was established. After tumor formation, PHPO was injected daily for treatment, and the tumor size was observed. Results · Compared to the control group, PHPO significantly inhibited the cell viability of A549 cells and induced apoptosis of them, and the IC50 value of 24 h is 44.23 μmol/L. PHPO blocked the cell cycle in the G1 phase significantly. The migration capacity of PHPO-treated cells was decreased. The mRNA levels of Bax and P21 were up-regulated in PHPO-treated group, and the mRNA lever of Bcl-2 was down-regulated (P<0.05). PHPO increased the phosphorylation levels of p38, ERK and JNK. Injection of PHPO could significantly inhibit the growth of tumor in the xenograft model compared to the control group (P<0.05). Conclusion · PHPO can induce the apoptosis and inhibit the proliferation of A549 cells, block the cell cycle in the G1 phase and decrease the migration ability of A549 cells significantly. The mechanism may be related to the activation of MAPK signaling pathway by PHPO and the increase of phosphorylation of p38, ERK and JNK.