Subject(s)
Gene Expression , Interleukin-8/metabolism , MAP Kinase Signaling System , Matrix Metalloproteinase 9/genetics , Pseudomonas Infections/genetics , RNA, Messenger/metabolism , Anthracenes/pharmacology , Bronchi , Cells, Cultured , Epithelial Cells , Flavonoids/pharmacology , Humans , Interleukin-8/genetics , MAP Kinase Signaling System/drug effects , Matrix Metalloproteinase 2/genetics , Matrix Metalloproteinase 2/metabolism , Matrix Metalloproteinase 9/metabolism , Mitogen-Activated Protein Kinase Kinases/metabolism , NF-kappa B/antagonists & inhibitors , NF-kappa B/genetics , Phosphorylation , Protein Kinase Inhibitors/pharmacology , Pseudomonas Infections/metabolism , Pseudomonas aeruginosa , Pyrrolidines/pharmacology , Thiocarbamates/pharmacologyABSTRACT
The pharmacology and clinical application of traditional Chinese medicine has been extensively documented. We have used an in vitro model system, PC12 cells, to demonstrate the presence of neuroactive compounds in Ganoderma lucidum (lingzhi). Ganoderma extract induced the neuronal differentiation of PC12 cells and prevented nerve growth factor-dependent PC12 neurons from apoptosis. Moreover, these effects of ganoderma might be mediated via the ras/extracellular signal-regulated kinase (Erk) and cAMP-response element binding protein (CREB) signaling pathways, as demonstrated by the phosphorylation of Erk1, Erk2 and CREB. Thus, our data not only present the first evidence of the presence of neuroactive compounds that mediate the neuronal differentiation and neuroprotection of the PC12 cells, but also reveal the potential signaling molecules involved in its action.
Subject(s)
Cell Differentiation/drug effects , Drugs, Chinese Herbal/pharmacology , Mitogen-Activated Protein Kinases/metabolism , Neurons/drug effects , Neurons/enzymology , Animals , Apoptosis/drug effects , Cell Division/drug effects , Cyclic AMP Response Element-Binding Protein/metabolism , Dose-Response Relationship, Drug , In Situ Nick-End Labeling , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3 , Nerve Growth Factor/pharmacology , Neurofilament Proteins/biosynthesis , Neurons/cytology , PC12 Cells , Pheochromocytoma/metabolism , Phosphorylation/drug effects , Rats , Receptor, trkA/metabolism , Reishi/chemistry , Signal TransductionABSTRACT
When treated with retinoic acid (RA), a human embryonal carcinoma (EC) cell line, NTera2 cl.D/1 (NT2), differentiates into several morphologically distinct cell types, which include terminally differentiated postmitotic central nervous system (CNS) neurons. Accumulating evidence has demonstrated the significant potential of NT2 cells in studies related to cancer therapy and neurodegenerative diseases. However, preparation of enriched NT2 neurons often requires a lengthy period (ca. five weeks) and depends largely on tedious techniques similar to those used for primary neuronal cultures. Here, we report a rapid protocol for the preparation of these human CNS neurons. Using the method of cell aggregation, enriched NT2 neurons can be obtained in approximately two weeks. We also demonstrated that cell aggregation reduced the time normally required for the induction of neuronal differentiation, as revealed by the early expression of neuronal markers. The period of RA treatment could also be reduced if NT2 cells were maintained as aggregates for a sufficient period of time. Taken together, our findings demonstrated that cell aggregation promoted RA-induced neuronal differentiation of NT2 cells and provided a rapid protocol for the efficient production of NT2 neurons. The ability to produce large quantities of human CNS neurons should facilitate future use of these neurons for basic research and applications in cell therapy.
