[Show abstract][Hide abstract] ABSTRACT: The ketogenic diet has long been used to treat epilepsy, but its mechanism is not yet clearly understood. To explore the potential mechanism, we analyzed the changes in gene expression induced by the ketogenic diet in the rat kainic acid (KA) epilepsy model.
KA-administered rats were fed the ketogenic diet or a normal diet for 4 weeks, and microarray analysis was performed with their brain tissues. The effects of the ketogenic diet on cathepsin E messenger ribonucleic acid (mRNA) expression were analyzed in KA-administered and normal saline-administered groups with semi-quantitative and real-time reverse transcription polymerase chain reaction (RT-PCR). Brain tissues were dissected into 8 regions to compare differential effects of the ketogenic diet on cathepsin E mRNA expression. Immunohistochemistry with an anti-cathepsin E antibody was performed on slides of hippocampus obtained from whole brain paraffin blocks.
The microarray data and subsequent RT-PCR experiments showed that KA increased the mRNA expression of cathepsin E, known to be related to neuronal cell death, in most brain areas except the brain stem, and these increases of cathepsin E mRNA expression were suppressed by the ketogenic diet. The expression of cathepsin E mRNA in the control group, however, was not significantly affected by the ketogenic diet. The change in cathepsin E mRNA expression was greatest in the hippocampus. The protein level of cathepsin E in the hippocampus of KA-administered rat was elevated in immunohistochemistry and the ketogenic diet suppressed this increase.
Our results showed that KA administration increased cathepsin E expression in the rat brain and its increase was suppressed by the ketogenic diet.
Yonsei medical journal 09/2010; 51(5):653-60. DOI:10.3349/ymj.2010.51.5.653 · 1.29 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In this study, we investigate the ability of reversine to stimulate adipocyte differentiation and its effect on cellular signaling pathways associated with adipocyte differentiation. Our data show that reversine treatment of 3T3-L1 cells under differentiation conditions synergistically enhances adipocyte differentiation and the expression of adipogenic marker genes such as aP2, PPAR-gamma, resistin, C/EBPalpha, and adiponectin. In parallel, reversine treatment leads to a selective downregulation of Akt and p70(s6k) signaling pathways, but not the ERK pathway. Furthermore, reversine stimulation of adipocyte differentiation seems to be quite different from troglitazone's action, because reversine treatment does not induce the transcriptional activation of PPAR-gamma and troglitazone does not affect the Akt and p70(s6k) signaling pathways. Taken together, our data clearly demonstrate the ability of reversine to stimulate adipocyte differentiation, which is independent of the Akt and p70(s6k) signaling pathways.
Biochemical and Biophysical Research Communications 07/2007; 358(2):553-8. DOI:10.1016/j.bbrc.2007.04.165 · 2.30 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Overexpression of P-glycoprotein (P-gp) is one of the major obstacles to successful cancer chemotherapy. In this study, we examined the ability of 4-chloro-N-(3-((E)-3-(4-hydroxy-3-methoxyphenyl)acryloyl)phenyl)benzamide (C-4) to reverse multidrug resistance (MDR) in P-gp expressing KBV20C cells. Treatment of KBV20C cells with C-4 led to a dramatic increase in paclitaxel- or vincristine-induced cytotoxicity without any cytotoxicity by itself. In parallel, C-4 treatment caused an increase in apoptotic cell death by paclitaxel or vincristine. Furthermore, C-4 treatment significantly increases in intracellular accumulation of fluorescent P-gp substrate rhodamine 123, indicating that C-4 treatment leads to reversal of the MDR phenotype resulting from an increased accumulation of anticancer drugs by inhibiting drug efflux function of P-gp. This notion is further supported by the observation that C-4 treatment potentiates paclitaxel-induced G(2)/M arrest of the cell cycle. In addition, the drug efflux function of P-gp was reversibly inhibited by C-4 treatment, while the expression level of P-gp was not affected. Collectively, our results describe the potential of C-4 to reverse the P-gp-mediated MDR phenotype through reversible inhibition of P-gp function, which may make it an attractive new agent for the chemosensitization of cancer cells.
Biochemical and Biophysical Research Communications 04/2007; 355(1):136-42. DOI:10.1016/j.bbrc.2007.01.117 · 2.30 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Histone deacetylase (HDAC) inhibitors are appreciated as one of promising anticancer drugs, but they exert differential responses depending on the cell type. We recently reported the critical role of NF-kappaB as a modulator in determining cell fate for apoptosis in response to an HDAC inhibitor. In this study, we investigate a possible signaling pathway required for NF-kappaB activation in response to the HDAC inhibitor apicidin. Treatment of HeLa cells with apicidin leads to an increase in transcriptional activity of NF-kappaB and the expression of its target genes, IL-8 and TNF-alpha. TNF-alpha expression by apicidin is induced at earlier time points than NF-kappaB activation or IL-8 expression. In addition, our data show that the early expression of TNF-alpha does not lead to activation of NF-kappaB, because disruption of TNF-alpha activity by a neutralizing antibody does not affect nuclear translocation of NF-kappaB, IkappaBalpha degradation or reporter gene activation by apicidin. However, this activation of NF-kappaB requires the PI3K and PKC signaling pathways, but not ERK or JNK. Furthermore, apicidin activation of NF-kappaB seems to result from HDAC1 inhibition, as evidenced by the observation that overexpression of HDAC1, but not HDAC2, 3 or 4, dramatically inhibits NF-kappaB reporter gene activity. Collectively, our results suggest that activation of NF-kappaB signaling by apicidin requires both the PI3K/PKC signaling pathways and HDAC1, and functions as a critical modulator in determining the cellular effect of apicidin.
Biochemical and Biophysical Research Communications 10/2006; 347(4):1088-93. DOI:10.1016/j.bbrc.2006.06.196 · 2.30 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In order to develop new anti-photoaging agents, we examined the inhibitory effects of 29 seaweed extracts on transcriptional activities of NF-kappaB and AP-1, and MMP-1 expression. The extracts from 3 species of Alariaceae, Eisenia bicyclis, Ecklonia cava and Ecklonia stolonifera, have showed strong inhibition of both NF-kappaB and AP-1 reporter activity, which were well correlated with their abilities to inhibit MMP-1 expression. In addition, MMP-1 expression was dramatically attenuated by treatment with eckol or dieckol which were purely isolated from E. stolonifera, indicating that these compounds are active principles to inhibit MMP-1 expression in human dermal fibroblasts. Taken together, our data demonstrate the inhibitory effect of eckol and dieckol from Ecklonia species on MMP-1 expression in human dermal fibroblasts and provide a possibility to develop as an agent for the prevention and treatment of skin aging.
[Show abstract][Hide abstract] ABSTRACT: Treatment with ergolide, a sesquiterpene lactone from Inula britannica var chinensis, caused the induction of apoptosis in Jurkat T cells, which was confirmed by DNA fragmentation, caspase-3 activation and cleavage of poly(ADP-ribose) polymerase in response to ergolide. Furthermore, mitochondrial dysfunction appeared to be associated with ergolide-induced apoptosis, because Bax translocation and cytochrome c release were stimulated by ergolide. In parallel, the nuclear factor-kappaB (NF-kappaB) signaling pathway was significantly inhibited by ergolide, which was accompanied by down-regulation of cell survival molecules, such as X-chromosome-linked inhibitor of apoptosis and Bcl-2. In addition, the JNK signaling pathway was involved in ergolide-induced apoptosis. Collectively, our results identified a new mechanism for the anti-cancer property of ergolide, attributable to the induction of apoptosis through down-regulation of cell survival signal molecules resulting from inhibition of the NF-kappaB signaling pathway.
Journal of Pharmacy and Pharmacology 01/2006; 57(12):1591-7. DOI:10.1211/jpp.57.12.0009 · 2.26 Impact Factor