PURPOSE: Several studies have proven that EGCG, the primary green tea catechin, and glucosamine-6-phosphate (PGlc) reduce triglyceride contents in 3T3-L1 adipocytes. The objective of this study is to evaluate the combination effect of EGCG and PGlc on decline of accumulated fat in differentiated 3T3-L1 adipocytes. METHODS: EGCG and PGlc were administered for 6 day for differentiation of 3T3-L1 adipocytes. Cell viability was measured using the CCK assay kit. In addition, TG accumulation in culture 3T3-L1 adipocytes was investigated by Oil Red O staining. We examined the expression level of several genes and proteins associated with adipogenesis and lipolysis using real-time RT-PCR and Western blot analysis. A flow cytometer Calibar was used to assess the effect of EGCG and PGluco on cell-cycle progression of differentiating 3T3-L1 cells. RESULTS: Intracelluar lipid accumulation was significantly decreased by combination treatment with EGCG 60 microM and PGlc 200 microg/m compared with control and EGCG treatment alone. In addition, use of combination treatment resulted in directly decreased expression of PPARgamma, C/EBPalpha, and SREBP1. In addition, it inhibited adipocyte differentiation and adipogenesis through downstream regulation of adipogenic target genes such as FAS, ACSL1, and LPL, and the inhibitory action of EGCG and PGlc was found to inhibit the mitotic clonal expansion (MCE) process as evidenced by impaired cell cycle entry into S phase and the S to G2/M phase transition of confluent cells and levels of cell cycle regulating proteins such as cyclin A and CDK2. CONCLUSION: Combination treatment of EGCG and PGlc inhibit-ed adipocyte differentiation through decreased expression of genes related to adipogenesis and adipogenic and cell cycle arrest in early stage of adipocyte differentiation.
3T3-L1 Cells ; Adipocytes* ; Adipogenesis* ; Blotting, Western ; Catechin ; Cell Cycle Checkpoints* ; Cell Cycle* ; Cell Survival ; Cyclin A ; Lipolysis ; Phase Transition ; PPAR gamma ; S Phase ; Tea ; Triglycerides

Apoptosis ; Biological Availability ; Blotting, Western ; Caspase 3 ; Caspase 9 ; Cell Death ; Cell Survival ; Cytochromes c ; Ellagic Acid ; Hydrolyzable Tannins ; Insurance Benefits ; Neurodegenerative Diseases ; Neurons ; Neuroprotective Agents ; Oxidative Stress ; p38 Mitogen-Activated Protein Kinases ; Phosphorylation ; Protein Kinases ; Reactive Oxygen Species ; Sincalide

PURPOSE: Many studies have suggested that neuronal cells protect against oxidative stress-induced apoptotic cell death by polyphenolic compounds. We investigated the neuroprotective effects and the mechanism of action of Momordica charantia ethanol extract (MCE) against H₂O₂-induced cell death of human neuroblastoma SK-N-MC cells. METHODS: The antioxidant activity of MCE was measured by the quantity of total phenolic acid compounds (TPC), quantity of total flavonoid compounds (TFC), and 2,2-Diphenyl-1-pycrylhydrazyl (DPPH) radical scavenging activity. Cytotoxicity and cell viability were determined by CCK-8 assay. The formation of reactive oxygen species (ROS) was measured using 2,7-dichlorofluorescein diacetate (DCF-DA) assay. Antioxidant enzyme (SOD-1,2 and GPx-1) expression was determined by real-time PCR. Mitogen-activated protein kinases (MAPK) pathway and apoptosis signal expression was measured by Western blotting. RESULTS: The TPC and TFC quantities of MCE were 28.51 mg gallic acid equivalents/extract g and 3.95 mg catechin equivalents/extract g, respectively. The IC₅₀ value for DPPH radical scavenging activity was 506.95 µg/ml for MCE. Pre-treatment with MCE showed protective effects against H₂O₂-induced cell death and inhibited ROS generation by oxidative stress. SOD-1,2 and GPx-1 mRNA expression was recovered by pre-treatment with MCE compared with the presence of H₂O₂. Pre-treatment with MCE inhibited phosphorylation of p38 and the JNK pathway and down-regulated cleaved caspase-3 and cleaved PARP by H₂O₂. CONCLUSION: The neuroprotective effects of MCE in terms of recovery of antioxidant enzyme gene expression, down-regulation of MAPK pathways, and inhibition apoptosis is associated with reduced oxidative stress in SK-N-MC cells.
Apoptosis ; Blotting, Western ; Caspase 3 ; Catechin ; Cell Death ; Cell Survival ; Down-Regulation ; Ethanol ; Gallic Acid ; Gene Expression ; Humans* ; Hydrogen* ; MAP Kinase Signaling System ; Mitogen-Activated Protein Kinases ; Momordica charantia* ; Momordica* ; Neuroblastoma* ; Neurons ; Neuroprotective Agents* ; Oxidative Stress ; Phenol ; Phosphorylation ; Reactive Oxygen Species ; Real-Time Polymerase Chain Reaction ; RNA, Messenger ; Sincalide

Dens in dente is a developmental anomaly resulting from infolding of the enamel organ into dental papilla prior to calcification of dental tissue. The pulpal tissue of the tooth can be vulnerable for bacterial invasion through direct exposure to the oral cavity or through defective enamel and dentin of the infolding part, thereby increasing the possibility of pulpal necrosis and subsequent apical periodontitis. Treatment planning of teeth with dens invaginatus may be difficult due to the complex root canal morphology. Therefore, thorough knowledge of anatomical variations of dens invaginatus is of great importance for proper treatment planning. The focus of this case report is on Oehler’s type II and III dens invaginatus. The infolding of type III dens invaginatus extends beyond the crown and CEJ. Bacterial invasion through the infolding can easily cause inflammation of the pulpal and periradicular tissue. This case report presents endodontic treatment of type II and III dens invaginatus with the aid of CBCT.