Résumé
Platycodin D (PD), a triterpenoid saponin isolated from Platycodonis Radix, is a famous Chinese herbal medicine that has been shown to have anti-proliferative effects in several cancer cell lines. The aim of this study was to determine the changes in cellular proteins after the treatment of hepatocellular carcinoma HepG2 cells with PD using proteomics approaches. The cell viability was determined using the MTT assay. The proteome was analyzed by two-dimensional difference gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Western blot analysis was used to confirm the expression of changed proteins. Our results showed that PD inhibited the proliferation of HepG2 cells in concentration- and time-dependent manners. Sixteen proteins were identified to be up-regulated in PD-treated HepG2 cells, including ATP5H, OXCT1, KRT9, CCDC40, ERP29, RCN1, ZNF175, HNRNPH1, HSP27, PA2G4, PHB, BANF1, TPM3, ECH1, LGALS1, and MYL6. Three proteins (i.e., RPS12, EMG1, and KRT1) decreased in HepG2 cells after treatment with PD. The changes in HSP27 and PHB were further confirmed by Western blotting. In conclusion, our results shed new lights on the mechanisms of action for the anti-cancer activity of PD.
Sujets)
Humains , Antinéoplasiques d'origine végétale , Pharmacologie , Utilisations thérapeutiques , Apoptose , Technique de Western , Campanulaceae , Chimie , Carcinome hépatocellulaire , Traitement médicamenteux , Métabolisme , Prolifération cellulaire , Survie cellulaire , Cellules HepG2 , Tumeurs du foie , Traitement médicamenteux , Métabolisme , Phytothérapie , Extraits de plantes , Pharmacologie , Utilisations thérapeutiques , Protéome , Métabolisme , Protéomique , Saponines , Pharmacologie , Utilisations thérapeutiques , Triterpènes , Pharmacologie , Utilisations thérapeutiques , Régulation positiveRésumé
<p><b>OBJECTIVE</b>To discuss the intervention effect of ligustrazine on ox-LDL-induced foam cells from the perspective of reverse cholesterol transport.</p><p><b>METHOD</b>RAW264.7 cultured in vitro was induced with 20 mg x L(-1) ox-LDL to establish the foam cell model, and intervened with ligustrazine. The lipid accumulation in cells was observed by the oil red O dyeing. The changes in total cholesterol and cholesterol ester in the cells were detected with the colorimetric method. The fluorescent quantitative PCR and Western blot were used to detect the mRNA expressions of PPARgamma, LXRalpha and ABCA1.</p><p><b>RESULT</b>Ligustrazine could reduce total cholesterol and cholesterol ester in foam cells, inhibit the lipid accumulation, and increase the mRNA and protein expressions of PPARgamma, LXRalpha and ABCA1.</p><p><b>CONCLUSION</b>Ligustrazine can promote the reverse cholesterol transport by increasing the gene expressions of PPARgamma, LXRalpha and ABCA1.</p>
Sujets)
Animaux , Souris , Membre-1 de la sous-famille A des transporteurs à cassette liant l'ATP , Génétique , Métabolisme , Transport biologique , Lignée cellulaire , Cholestérol , Métabolisme , Médicaments issus de plantes chinoises , Pharmacologie , Cellules spumeuses , Métabolisme , Expression des gènes , Récepteur PPAR gamma , Génétique , MétabolismeRésumé
In the 1960s, modern science began involving the essence of heat syndrome, but there have still no in-depth systematic studies on pathological mechanisms of heat syndrome and action mechanisms of cold and cool herbs. In this study, the animal model with heat syndrome was set up by feeding herbs with hot property, and then cold and cool herbs was applied in the experimental therapy. The two-dimensional electrophoresis and mass spectrometry technologies were adopted to compare the liver mitochondria proteome of the rats of the heat syndrome model and the ones treated with cold and cool herbs, so as to discover specificity-related proteins after heat syndrome and treatment with cold and cool herbs.