• 대한본초학회지
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• 제27권4호
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• pp.17-23
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• 2012

Objectives : Schisandrae Fructus (SF) has traditionally been used to balance level of body fluid and to strengthen kidney function. It has been reported that the SF extract has antioxidant, hepatoprotective, neuroprotective and anticancer effects. This study investigated an antiproliferative effect of SF extract on PC-3 human prostate cancer cells and analyzed active ingredients of SF extract qualitatively and quantitatively. Methods : We examined the antiproliferative effect of SF extract with MTT assay, DAPI staining and annexin-V/7-AAD double staining. The active ingredients of SF extract were identified by using HPTLC and HPLC/DAD system. Results : SF-chloroform fraction inhibited growth of PC-3 cells and changed the morphology of nucleus in a dose dependent manner. A dose-dependent apoptotic cell death was also measured by flow cytometry analysis. It was analyzed that SF-chloroform fraction contained more schizandrin than other fractions by using HPTLC and HPLC/DAD system. Conclusions : These results suggest that SF extract and schizandrin may be a potential chemotherapeutic agent for the control of PC-3 human prostate cancer cells.

• BMB Reports
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• 제35권6호
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• pp.562-567
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• 2002

In order to find the cellular interaction factors of the Heliothis armigera nuclear polyhedrosis virus capsid protein VP39, a Heliothis armigera cell cDNA library was constructed. Then VP39 was used as bait. The host actin gene was isolated from the cDNA library with the yeast two-hybrid system. This demonstrated that VP39 could interact with its host actin in yeast. In order to corroborate this interaction in vivo, the vp39 gene was fused with the green fluorescent protein gene in plasmid pEGFP39. The fusion protein was expressed in the Hz-AM1 cells under the control of the Autographa californica multiple nucleopolyhedrovirus immediate early gene promoter. The host actin was labeled specifically by the red fluorescence substance, tetramethy rhodamine isothicyanete-phalloidin. Observation under a fluorescence microscopy showed that VP39, which was indicated by green fluorescence, began to appear in the cells 6 h after being transfected with pEGFP39. Red actin cables were also formed in the cytoplasm at the same time. Actin was aggregated in the nucleus 9 h after the transfection. The green and red fluorescence always appeared in the same location of the cells, which demonstrated that VP39 could combine with the host actin. Such a combination would result in the actin skeleton rearrangement.

• BMB Reports
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• 제41권12호
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• pp.863-867
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• 2008

CD320 has been recently discovered and reported as a follicular dendritic cell (FDC) protein. Although CD320 is known to enhance proliferation of germinal center (GC) B cells, little other information is available. In this study, we investigated its cellular distribution in the GC. Confocal microscopy of human tonsil sections revealed co-localization of CD320 with CD19 and CD38 but not with CD3 indicating that GC B cells expressed CD320 in addition to FDC. In purified GC B cells, CD320 expression was inhibited in the nucleus, membrane and cytoplasm. Reverse transcriptase-polymerase chain reaction confirmed CD320 mRNA expression in B cells. These finding indicate that CD320 is expressed in B cells in addition to FDC, and that its GC activity may be more complicated than previously thought.

• Molecules and Cells
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• 제22권3호
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• pp.285-290
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• 2006

The light sensing system in the eye directly affects the circadian oscillator in the mammalian suprachiasmatic nucleus (SCN). To investigate this relationship in the rat, we examined the circadian expression of clock genes in the SCN and eye tissue during a 24 h day/night cycle. In the SCN, rPer1 and rPer2 mRNAs were expressed in a clear circadian rhythm like rCry1 and rCry2 mRNAs, whereas the level of BMAL1 and CLOCK mRNAs decreased during the day and increased during the night with a relatively low amplitude. It seems that the clock genes of the SCN may function in response to a master clock oscillation in the rat. In the eye, the rCry1 and rCry2 were expressed in a circadian rhythm with an increase during subjective day and a decrease during subjective night. However, the expression of Opn4 mRNA did not exhibit a clear circadian pattern, although its expression was higher in daytime than at night. This suggests that cryptochromes located in the eye, rather than melanopsin, are the major photoreceptive system for synchronizing the circadian rhythm of the SCN in the rat.

• 대한한의학방제학회지
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• 제19권1호
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• pp.161-174
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• 2011

Objectives : The effects of Extraction after Mixture of Bupleuri Radix and Lycii Radicis Cortex on the change of the HPA-Axis system and the Catecholamic system was investigated. Methods : After performing the Forced Swimming Test(FST), the expressions of corticotropin releasing factor(CRF), c-Fos in the paraventricular nucleus(PVN), and tyrosine hydroxylase(TH) in the ventral tegmental area(VTA) and locus coeruleus(LC) were measured by immunohistochemical method. Results : The duration of immobility in FST was significantly decreased in A 100(Extraction after Mixture of Bupleuri Radix and Lycii Radicis Cortex, 100mg/kg) and A 400(Extraction after Mixture of Bupleuri Radix and Lycii Radicis Cortex, 400mg/kg)(p<0.001). The expression of CRF in the PVN was shown the tendency to reduce in A100 and A400. The expression of c-Fos in the PVN was shown the tendency to reduce in A100 and A400. The expression of TH in the VTA was shown the tendency to reduce mildly in A100 and A400. The expression of TH in the LC was significantly reduced in A400(p<0.001). And the dose dependent reduction tendency was shown, respectively. Conclusions : According to the results above mentioned, the immobility, c-Fos and CRF expression was reduced at lower dose and was increased at higher dose. Therefore there is contradictory effects on the HPA Axis system in accordance with the dose. But in the effects on the catecholaminergic system, it significantly reduced the expression of TH in the LC. It was validated that the effect on the catecholaminergic system was ruled by Bupleuri Radix rather than Lycii Radicis Cortex via mainly the noradernergic system.

• 감성과학
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• 제20권4호
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• pp.113-126
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• 2017

The purpose of the study was to find grey matter (GM) and white matter (WM) volume reduction in the brain reward system among patients with alcohol dependency. This study investigated regional GM and WM in chronic alcoholic patients, focusing primarily on the reward system, including principal components of the mesocorticolimbic reward circuit as well as cortical areas with modulating and oversight functions. Sixteen abstinent long-term chronic alcoholic men and demographically matched 16 healthy control men participated in the study. Morphometric analysis was performed on magnetic resonance brain scans using voxel-based morphometry (VBM)-diffeomorphic Anatomical Registration through Exponentiated Liealgebra (DARTEL). We derived GM and WM volumes from total brain and cortical and subcortical reward-related structures. Morphometric analyses that revealed the total volume of GM and WM was reduced and cerebrospinal fluid (CSF) was increased in the alcohol group compared to control group. The pronounced volume reduction in the reward system was observed in the GM and WM of the nucleus accumbens (NAc), GM of the amygdala, GM and WM of the hippocampus, WM of the thalamus, GM and WM of the insula, GM of the dorsolateral prefrontal cortex (DLPFC), GM of the orbitofrontal cortex (OFC), GM of the cingulate cortex (CC), GM and WM of the parahippocampal gyrus in the alcohol group. We identified volume reductions in WM as well as GM of reward system in the patients with alcohol dependency. These structural deficits in the reward system elucidate underlying impairment in the emotional and cognitive processing in alcoholism.

• 동의생리병리학회지
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• 제21권6호
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• pp.1483-1490
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• 2007

This study was carried out to investigate the effects of GCP treatment on the expression of NOS, c-fos, serotonin and substance P in central nerve system of monosodium iodoacetate(MIA)-induced osteoarthritic pain model. Arthritis was induced by injection of MIA(0.5 mg) into knee joint cavities of rats. Arthritic rats were divided into control(n=8) and treated(n=8) group. Control group was taken distilled water for 20 days. Treated group was taken extracts of GCP by oraly for same duration. Normal group(n=8) was infected with normal saline and was taken distilled water for 20 days. The numbers of NADPH-d positive cells in superficial dorsal horn of spinal cord of treated group($21{\pm}5$) was significantly (p<0.01) decreased compared with control($33{\pm}5$). The numbers of NADPH-d positive cell in dorsolateral periaqueductal gray matter of treated group($111{\pm}16$) was significantly(p<0.01) decreased compared with control($143{\pm}14$). The numbers of c-fos positive cells in dorsal periaqueductal gray matter of treated group($57{\pm}16$) was significantly(p<0.01) decreased compared with control($78{\pm}13$). The numbers of c-fos positive cells in paraventricular thalamic nucleus of treated group($60{\pm}15$) was significantly decreased compared with control($88{\pm}27$). The numbers of serotonin positive cells in median raphe nucleus of treated group($171{\pm}31$) was significantly(p<0.05) decreased compared with control($217{\pm}48$). On the basis of these results, we concluded that GCP treatment has inhibiting effects on the pain transmission in monosodium iodoacetate-induced osteoarthritic pain model in rat.

• The Korean Journal of Physiology
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• 제24권2호
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• pp.331-344
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• 1990

What makes glucose transport function sensitive to insulin in one cell type such as adipocyte, and insensitive in another such as liver cells is unresolved question at this time. Recently it is known that insulin stimulates glucose transport in adipocytes largely by redistributing transporter from the storage pool that is included in a low density microsomal fraction to plasma membrane. Therefore, insulin sensitivity may depend upon the relative distribution of gluscose transporters between the plasma membrane and in an intracellular storage compartment. In hepatocytes, the subcellular distribution of glucose transporter is less well documented. It is thus possible that the apparent insensitivity of the hepatocyte system could be either due to lack of the constitutively maintained, intracellular storage pool of glucose transporter or lack of insulin-mediated transporter translocation mechanism in this cell. In this study, I examined if any intracellular glucose transporter pool exists in hepatocytes and this pool is affected by insulin. The results obtained summarized as followings: 1) Distribution of subcellular fractions of hepatocyte showed that there are $24.9{\pm}1.3%$ of plasma membrane, $36.9{\pm}1.7%$ of nucleus-mitochondria enriched fraction, $23.5{\pm}1.2%$ of lysosomal fraction, $9.6{\pm}1.0%$ of high density microsomal fraction and $4.9{\pm}0.5%$ of low density microsomal fraction. 2) In adipocyte, there were $29.9{\pm}2.6%$ of plasma membrane, $19.4{\pm}1.9%$ of nucleus-mitochondria enriched fraction, $26.7{\pm}1.8%$ of high density microsomal fraction and $23.9{\pm}2.1%$ of low density microsomal fraction. 3) Surface labelling of sodium borohydride revealed that plasma membrane contaminated to lysosomal fraction by $26.8{\pm}2.8%$, high density microsomal fraction by $8.3{\pm}1.3%$ and low density microsomal fraction by $1.7{\pm}0.4%$ respectively. 4) Cytochalasin B bound to all of subcellular fractions with a Kd of $1.0{\times}10^{-6}M$. 5) Photolabelling of cytochalasin B to subcellular fractions occurred on 45 K dalton protein band, a putative glucose transporter and D-glucose inhibited the photolabelling. 6) Insulin didn't affect on the distribution of subcellular fractions and translocation of intracellular glucose transporters of hepatocytes. 7) HEGT reconstituted into hepatocytes was largely associated with plasma membrane and very little was found in low density microsomal fraction which equals to the native glucose transporter distribution. Insulin didn't affect on the distribution of exogeneous glucose transporter in hepatocytes. From the above results it is concluded that insulin insensitivity of hepatocyte may due to lack of intracellular storage pool of glucose transporter and thus intracellular storage pool of glucose transporter is an essential feature of the insulin action.

• 대한약침학회지
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• 제7권1호
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• pp.53-61
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• 2004

Substantial evidence suggests that behavioral and reinforcing effects of cocaine can be mediated by the mesolimbic dopaminergic system. It has been shown that repeated injections of cocaine produce increase in locomotor activity, expression of the immediate-early gene, c-fos in the nucleus accumbens (NAc), which was one of the main dopaminergic terminal areas. Herbal-acupuncture as a therapeutic intervention has been widely used for the treatment of many functional disorders such as drug abuse. Coptidis Rhizoma (CR) and its main component, berberine (BER) were selected as herbal medicine of herbal-acupuncture. Both medicines have been known to have the therapeutic effect on the central nervous system. In order to investigate the effects of CR and BER herbalacupuncture at shenmen (HT7) point (CR/H and BER/H) on the cocaine-induced behavioral sensitization, the influence of CR/H and BER/H on repeated cocaine-induced locomotor activity, the change of c-Fos expression in the brain by immunohistochemistry were examined. Male SD rats were given CR/H (0.4mg/kg) and BER/H (0.1mg/kg) 30 min before daily injections of cocaine hydrochloride (15mg/kg. i.p.) 10 days. After 3 days withdrawal, rats received a challenge injection of cocaine (15mg/kg, i.p.). Systemic challenge with cocaine produced much larger increased locomotor activity, accumbal Fos-like immunoreactivity in the NAc. Pretreatment with CR/H and BER/H significantly inhibited cocaine-induced locomotor activity, the change of c-Fos expression in the rats. Our data demonstrated that the inhibitory effects of cocaine-induced behavioral sensitization by CR/H and BER/H were closely associated with the reduction of presynaptic dopamine release in the NAc. These results suggest that CR/H and BER/H can be effectively applied to cocaine addiction.

• 대한한의학방제학회지
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• 제18권1호
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• pp.133-144
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• 2010

Object : The effects of Rehmanniae Radix(RR) on HPA Axis system and Catecholaminergic system was investigated. Methods : we performed the Forced Swimming Test(FST). Also the expression of Corticotropin -Releasing Factor(CRF), c-Fos and Tyrosine Hydorxylase(TH) was measured by immunohistochemical method at Paraventricular Nucleus(PVN), Locus Coeruleus(LC) and Ventral Tegmental Area(VTA). Results : 1. The duration of immobility in the Forced Swimming Test was significantly decreased in the Rehmanniae Radix 400 mg/kg groups in comparison with the control group(p<0.05). 2. Comparing to the control group, CRF expression was significantly decreased in the Rehmanniae Radix 400 mg/kg treated group in comparison with the control group(p<0.01). 3. c-Fos expression was significantly decreased at PVN in the Rehmanniae Radix 100 mg/kg treated group in comparison with the control group(p<0.01). 4. TH expression was significantly increased at LC in the Rehmanniae Radix 100 mg/kg and 400 mg/kg treated group in comparison with the control group(p<0.001). 5. TH expression was significantly decreased at VTA in the Rehmanniae Radix 100 mg/kg and 400 mg/kg treated group in comparison with the control group(p<0.001). Conclusion : According to the results, it can be considered that Rehmanniae Radix has antidepressant effect by showing the reduction of immobility in FST through the decreased expression of CRF, c-Fos in PVN and TH in VTA.