• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.2
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• pp.331-337
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• 2001

The effect of Phellinus linteus methanol extract on the lipid metabolism in the liver of rat was investigated in this study. Rats were randomly divided into 6 groups including the control, $CCl_4$and high fat group plus sub-groups with Phellinus linteus methanol extract administration. Methanol extracts of Phellinus linteus were fed 50mg/kg B.W daily via drinking water. A 1.2mL of $CCl_4/kg$ body weight was administered by oral intubation twice a week for total six times. The administration of $CCl_4$ increased total cholesterol, TG, LDL and LDL-phospholipid. However, the level of liver cholesterol and triglycerides were significantly decreased while HDL-cholesterol was increased by the extract feeding. The activities of GOT, GPT, AP and LDH were greatly enhanced by the extract feeding. Electronmicrograph showed that $CCl_4$ treatment deteriorated the structure of cytoplasmic matrix with its uneven distribution. However, the extract treatment reconstituted the damaged cytoplasm and stimulated mitochondriagenesis. From these results, it was suggested that Phellinus linteus can help to recover the damaged liver function and further may help to prevent senescence diseases such as fatty liver, hypertension and hyperlipidemia.

• The Korean Journal of Pain
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• v.22 no.1
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• pp.16-20
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• 2009

Background: Zaprinast is an inhibitor of phosphodiesterase 5, 6 and 9. Phosphodiesterase inhibitors could produce anti-nociceptive effects by promoting the accumulation of cGMP. We hypothesized that intrathecal zaprinast could attenuate the allodynia induced by chronic constriction injury of the sciatic nerve in rat. Methods: Sprague-Dawley rats were prepared with four loose ligations of the left sciatic nerve just proximal to the trifurcation into the sural, peroneal and tibial nerve branches. Tactile allodynia was measured by applying von Frey filaments to the lesioned hindpaw. The thresholds for the withdrawal responses were assessed. Zaprinast ($3-100{\mu}g$) was administered intrathecally by the direct lumbar puncture method to obtain the dose-response curve and the 50% effective dose ($ED_{50}$). Measurements were taken before and 15, 30, 45, 60, 90, 120, and 180 min after the intrathecal doses of zaprinast. The side effects were also observed. Results: Intrathecal zaprinast resulted in a dose-dependent antiallodynic effect. The maximal effects occurred within 15-30 min and then they gradually decreased down to the baseline level over time in all the groups. There was a dose dependent increase in the magnitude and duration of the effect. The $ED_{50}$ value was $17.4{\mu}g$ (95% confidence intervals; $14.7-20.5{\mu}g$). No severe motor weakness or sedation was observed in any of the rats. Conclusions: Intrathecally administered zaprinast produced a dose-dependent antiallodynic effect in the chronic constriction injury neuropathic pain model. These findings suggest that spinal phosphodiesterase 5, 6 and 9 may play an important role in the modulation of neuropathic pain.

• Journal of Pharmaceutical Investigation
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• v.38 no.1
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• pp.31-38
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• 2008

The objective of this work is to study transdermal delivery of levodopa using iontophoresis and evaluate various factors which affect the transdermal transport. Levodopa is unstable in aqueous solution, and, in order to establish a stable condition for levodopa for the duration of experiment, we investigated the stability of levodopa in aqueous solutions of different pHs with/without the addition of dextrose or the application of current. Using stable aqueous solution, we have studied the effect of pH, polarity and penetration enhancer (ethanol) on transdermal flux and compared the results. We also investigated the iontophoretic flux from hydroxypropyl cellulose (HPC) hydrogel. In vitro flux study was performed at $33^{\circ}C$, using side-by-side diffusion cell. Full thickness hairless mouse skin and rat skin were used for this work. Current densities applied were 0.4 or $0.6mA/cm^2$ and current was off after 6 hour application. Stability study showed that levodopa solution with a pH 2.5 or 4.5 maintained the initial concentration of levodopa for 24 hours with the addition of 5% dextrose. However, at pH 9.5, levodopa was unstable and 30 to 40% of levodopa degraded within 24 hours, even with the addition of 5% dextrose. Hydrogel swollen with dextrose added levodopa solution maintained about 97% of the initial concentration of levodopa for 13 days, when stored in $4^{\circ}C$. The application of current did not affect the stability of levodopa in hydrogel. Flux study from levodopa solution with pH 2.5 showed that cathodal delivery of levodopa was higher than passive or anodal delivery. When the pH of the donor solution was 4.5, anodal delivery of levodopa was higher than passive or cathodal delivery. These results seem to indicate that electroosmosis plays more dominant role than electrorepulsion in the flux of levodopa at pH 2.5, and the reverse situation applies for pH 4.5. The passive flux was unexpectedly high for the ionized levodopa. Similar to the results from aqueous solution, cumulative amount of levodopa transported trom HPC hydrogel by cathodal delivery was significantly higher than passive or anodal delivery. The treatment of 70% ethanol cotton ball by scrubbing increased passive, anodal and cathodal flux, with the largest increase for anodal flux. These results indicate that iontophoretic delivery of zwitterion such as levodopa is much complicated than that can be expected from small ionic molecules with single charge. The results also indicate that the balance between electroosmosis and electrorepulsion plays a very important role in the transport through skin.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.6
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• pp.713-717
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• 1999

In order to utilize protein hydrolysate produced from hoki (Johnius Belengeri) frame among many different fish processing wastes, hoki frame peptide (PHFP) and phosphorylated hoki frame peptide (PHFP) were prepared, and their calcium absorption accelerating effects were investigated in comparison to control and casein phosphopeptide (CPP). In in vitro experiment, HFP and PHFP inhibited calcium phosphate formation as high as 1.5-fold and 2.5-fold, respectively, comparing to control, In addition, the inhibition rate of calcium phosphate precipitation as increasing concentrations of HFP and PHFP was risen and was similar to that of CPP at 2.0 mg/ml of PHFP concentration, In in vivo experiment using the rats, the groups fed HFP and PHFP indicated significantly increased calcium content in the femur. In particular, the calcium content in the small intestine of the rat fed PHFP was higher than that of control group by approximately $60\%$.

• Journal of Periodontal and Implant Science
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• v.32 no.3
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• pp.501-521
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• 2002

The purpose of this study is to evaluate the influences of the bone morphogenetic protein (BMP) on the healing of periodontal ligament and alveolar bone after replantation of tooth, and to examine the possibility of its clinical application. 45 Sprague Dawley rats weighted about 100 gram were divided into 3 experimental groups by different dose of BMP. All the upper right and left 1st molar were extracted after 5 days feeding of 0.4% ${\beta}$-aminopropionitrile, and right molar were used as experimental group and left molar were used as control group. The root surface of experimental molar were treated with 25,50 and l00ng/ml of human recombinant Bone morphogenetic protein-4 (rh-BMP-4) with micropipet, and 1M Sodium hypochloride were used on control root surface. All the experimental animals were sacrificed as 1, 2, 4, 7 and 14 days after autoreplantation of upper 1st molar into their own position. The maxilla were disected included both side of 1st molar. The collected tissue were processed from demineralization to paraffin embeding as usual procedure, and the specimens were prepared with Hematoxylin-Eosin stain for the light microscopic evaluation. The results of this study were as follows : 1. There was no significant differences between control and experimental site on 1 and 2 days after replantation of tooth. In the case of 4th days, the evidence of tissue regeneration were observed on experimental site to compare the controls. New osteoid were revealed on high concentration of BMP at 7 days after replantation, and it became more obvious at 14 days, 2. The effect of the rh-BMP-4 coated on root surface was revealed slight influences for the prolifertion of cells of periodontium and tissue regeneration as dose-dependent pattern. 3. Bony ankylosis was observed between alveolar bone and root surface due to the remarkable amount of osteoid formation on the 14 days after replantation of root. In the conclusion, it was suggested that topical application of the rhBMP-4 on the root surface has influence on the periodontal ligament and alveolar bone. The application method of BMP on the root should be designed with calculation of proper concentration.

• Journal of Periodontal and Implant Science
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• v.30 no.2
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• pp.287-305
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• 2000

The purpose of this study is to evaluate the effects of bioresorbable membranes in guided bone regeneration of streptozotocin induced diabetic rats. 50 Sprague-Dawley rats were randomly categorized into 4 groups: Group 1 & 2 had 10 normal rats each and group 3 & 4 included 15 streptozotocin induced diabetic rats each. Defect measuring 7mm in diameter was formed on every rat calvarium. No membrane was used in groups 1 & 3 and membranes were used in groups 2 & 4. The rates were sacrificed at 2 and 4 weeks after defect formation. Routine histological specimens were prepared. Masson-trichrome and HE stain were done before light microscopy. Guided regenerative potential was evaluated by measuring the amount of new bone formation in the calvarial defect by histomorphometry. Following results were obtained. 1. New bone formation in the diabetic groups was significantly less that than in the normal groups regardless of membrane use(p<0.05). 2. In the comparison of new bone formation in the normal groups, membrane group showed significantly more bone formation(p<0.1). 3. When the amount of new bone formation was compared in the diabetic groups, more bone was formed in the membrane groups but the difference was not statistically significant.4. In the normal groups the amount of new bone formation was significantly greater at 4 weeks compared to that at2 weeks(p<0.05) but amount of bone regeneration at 4 weeks was not significantly greater than that at 2 weeks in both diabetic groups.

• Journal of Periodontal and Implant Science
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• v.29 no.4
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• pp.751-765
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• 1999

Several growth factors and polypeptidesare not commonly yet used for regenerators of bone tissue or alveolar bone because of the insufficiency of studies on their side effects, genetic engineering for mass production and stability for clinical application. Recently, many natural medicines, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, anti-inflammatory and regenerative potential of periodontal tissues. Olibanum, Myrrha, Phlomis Radix, and Cimicifugae Rhizoma have been traditionally used as a drug for treatment of bone disease in oriental medicine. The objective of this study was to examine the ability of alkaline phosphatase(ALP) synthesis of rat calvarial osteoblast(MC3T3-E1) when several natural medicines were supplemented. MC3T3-E1 cells were cultured with ${\alpha}$-MEM(negative control), dexamethasone(positive control), and each natural medicines for 3 and 5 days. And then ALP synthesis was measured by spectrophotometer for enzyme activity and by naphthol AS-BI staining for morphometry. All of the natural medicines induced higher activity of ALP synthesis than the negative controls. Especially Olibanumind uced the higher activity than the positive controls (p<0.05). In the aspects of culturing time, except Cimicifugae Rhizoma, the natural medicines induced higher activity of ALP synthesis at 5 days than at 3 days (p<0.05). In morphometry, all of the natural medicines showed statistical significance compared to the negative control (p<0.05). Myrrha a n d Phlomis Radix showed larger positively stained area at 5days than at 3 days, whereas the others did not showed the difference between at 5 and at 3 days(p<0.05). These results indicate that several natural medicines have an inducing ability of ALP synthesis in MC3T3-E1 cells.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2002.10a
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• pp.168-168
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• 2002

To establish a test protocol for the rodent 20-day thyroid/pubertal assay, flutamide and diethylstilbestrol (DES) were administered to intact male Sprague-Dawley rats from postnatal day 33 for 20 days. Flutamide (1, 5, and 25 mg/kg/day) or DES (10, 20, and 40 ug/kg/day) was given once daily by oral gavage to immature male rats. Prepuce separation was significantly delayed in flutamide group and in DES group. One day after the last dose, the rats were killed and pituitary, thyroid, and reproductive organs were removed and weighed. Flutamide treatment resulted in a significant reduction in the weights of epididymides, ventral prostate, seminal vesicles plus coagulating glands and fluid (SVCGF), levator ani. bulbocarvenus muscles (LABC), Cowper's glands, and glans penis. The weight of adrenal glands decreased at % mg/kg/day, while testes and any other organ weights were unaffected. No microscopic changes were observed in the thyroid glands. Serum levels of testosterone wert significantly increased in the flutamide-treated groups and serum levels of estradiol were also increased. A significant reduction in the weights of testes, epididymides, ventral prostate, SVCGF, LABC, Cowpers glands, and glans penis of DES treated group. Serum testosterone and LH decreased significantly in DES group. Decrease of estradiol was observed, but not significant. These results indicate that flutamide and DES delay puberty in the male rat and its mode of action appears to be via altered secretion of steroids, which subsequently affect the development of the reproductive tract. (Supported by the grant from NITR/Korea FDA for Endocrine Disrupter Research.)

• Development and Reproduction
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• v.20 no.4
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• pp.315-329
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• 2016

Fas ligand (FasL) and its receptor Fas have been implicated in granulosa cell apoptosis during follicular atresia. Although interferon-gamma (IFN-${\gamma}$) is believed to be involved in the regulation Fas expression in differentiated granulosa or granulosa-luteal cells, the expression of this cytokine and its role in the regulation of the granulosa cell Fas/FasL system and apoptosis during follicular maturation have not been thoroughly investigated. In the present study, we have examined the presence of IFN-${\gamma}$ in ovarian follicles at different stage of development by immunohistochemistry and related their relative intensities with follicular expression of Fas and FasL, and with differences in granulosa cell sensitivity to Fas activation by exogenous agonistic Anti-Fas monoclonal antibody (Fas mAb). Although IFN-${\gamma}$ immunostaining was detectable in oocyte and granulosa cells in antral follicles, most intense immunoreactivity for the cytokine was observed in these cells of preantral follicles. Intense immunoreactivity for IFN-${\gamma}$ was most evident in granulosa cells of atretic early antral follicles where increased Fas and FasL expression and apoptosis were also observed. Whereas low concentrations of IFN-${\gamma}$ (10-100 U/mL) significantly increased Fas expression in undifferentiated granulosa cells (from preantral or very early antral follicles) in vitro, very higher concentrations (${\geq}1,000U/mL$) were required to up-regulate of Fas in differentiated cells isolated from eCG-primed (antral) follicles. Addition of agonistic Fas mAb to cultures of granulosa cells at the two stages of differentiation and pretreated with IFN-${\gamma}$ (100 U/mL) elicited morphological and biochemical apoptotic features which were more prominent in cells not previously exposed to the gonadotropin in vivo. These findings suggested that IFN-${\gamma}$ is an important physiologic intra-ovarian regulator of follicular atresia and plays a pivotal role in regulation of expression of Fas receptor and subsequent apoptotic response in undifferentiated (or poorly differentiated) granulosa cells at an early (penultimate) stage of follicular development.

• Reproductive and Developmental Biology
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• v.30 no.4
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• pp.247-253
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• 2006

Our previous research has identified granulin (grn) and p130 genes as sex steroid-inducible genes in the rat hypothalamus, which might be involved in sexual differentiation of the brain. Phthalate esters that are used as plasticizers and also found at low levels in foods such as dairy products are often mentioned as suspected endocrine disrupters. The purpose of the present study is to elucidate whether perinatal exposure to di-n-butyl phthalate (DBP), diisononyl phthalate (DINP) and di-2-ethylhexyl adipate (DEHA) affects hypothalamic sex steroid-inducible genes. The present study assessed the effects of perinatal exposure to DBP, DINP and DEHA on sex steroid hormones levels and hypothalamic gm and p130 mRNA expressions at postnatal day (PND) 3 and 7. Pregnant rats were fed a soy-free diet containing 20, 200, 2,000 and 10,000 ppm of DBP, 40, 400, 4,000 and 20,000 ppm of DINP, or 480, 2,400 and 12,000 ppm of DEHA from gestational day (GD) 15 to GD 3 or 7. At PND 3 and 7, perinatal exposure to these chemicals did not substantially affect serum concentrations of testosterone and estradiol. At PND 3, the expression of grn mRNA levels in males was decreased by DEHA, and that of p130 was decreased by DBP, DINP and DEHA, though the effects were not dose-dependent. At PND 7, the expression of gm gene in female pups was increased by higher doses of DBP and all the doses, except for 4,000 ppm, of DINP, while that in male pups decreased by 480 and 12,000 ppm of DEHA. Hypothalamic expression of p130 mRNA in males was increased by lower doses of DBP and all the doses of DINP, whereas that of females was decreased by 480 and 2,400 ppm of DEHA. These results suggest that these chemicals may affect the expression of gm and p130 genes by directly acting on the hypothalamus, thus leading to inappropriate expression of these genes.