• Journal of Periodontal and Implant Science
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• 제34권4호
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• pp.807-818
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• 2004

산화질소는 생리적 농도에서 세포내 신호전달자로 작용하지만 높은 농도에서는 세포독성을 일으킨다. 최근 치은 섬유아세포와 치주인대 섬유아세포는 산화질소 합성효소를 가지고 있고 세균의 lipopolysaccharide나 cytokine에 의해 대량의 높은 농도의 산화질소가 합성된다는 보고가 있음에도 지금까지 치은 조직에서 산화질소의 세포독성에 대한 연구는 아직 이루어 지지않고 있다. 본 연구는 사람의 치은 섬유아세포에서, 산화질소유도세포 고사기전을 밝히는데 목적이 있다. 세포 생장력은 MTT 방법으로 측정하였고, 세포의 형태적 변화는 Diff-Quick 염색법으로 조사하였다. Bcl-2 famly와 Fas 발현 정도는 RT-PCR 방법에 의해 확인하였으며, caspase-3, -8 와 -9의 활성은 spectrophotometer로 reactive oxygen species (ROS)는 형광분광계에 의해 측정되었다. 미토콘드리아에서 세포질로 분비된 cytochrome c는 western blot으로 조사하였다. 산화질소 유리제인 sodium nitroprusside (SNP) 처리는 사람 섬유아세포의 생존률을 시간과 농도 의존적으로 감소시켰고, 세포용적축소, 염색사 용축, DNA 절편화를 일으켰다. 또한, SNP 처리로 미토콘드리아에서 세포질로 유리되는 cytochrome c 양이 증가되었고, caspase-9 과 caspase-3 의 활성이 증가되었다. 한편, SNP 처리에 의해 death receptor 구성요소인 Fas 발현이 증가되었고, caspase-8의 활성이 증가되었다. Bcl-2 family 에 대한 RT-PCR 분석결과, 세포고사를 억제하는 Bcl-2 발현은 감소되었으나 세포고사를 자극하는 Bax와 Bid의 발현은 증가되었다. Soluble guanylate cyclase 억제제인 ODQ는 SNP에 의한 세포 생존율 감소를 차단하지 못했다. 따라서, 본 실험의 결과들은 사람 섬유아세포에서 산화질소유도 세포고사에 Bcl-2 family나 ROS가 매개하는 미토콘드리아 의존 및 death receptor 의존 세포고사기전이 관여함을 시사하였다.

• 동의생리병리학회지
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• 제23권4호
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• pp.799-804
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• 2009

Kamikaekyuk-tang(KMKKT), a formula of ten Oriental herbs, was orientally designed to promote vital energy, to remove blood stasis, and to decrease inflammation for treating cancers. KMKKT and its component had potent antiandrogen and androgen receptor activities in prostate cancer and also inhibited angiogenesis induced by basic fibroblast growth factor (bFGF) in human umbilical vein endothelial cells and suppressed the tumor growth in LLC-bearing mice, and liver metastasis of colon 26-L5 cancer cells, suggesting a potent cancer preventive agent. Nevertheless, there is no safety study of KMKKT before clinical trial so far. Thus, in the current study, we investigated the toxicity about ethanol-extracted KMKKT. Male and female Spraque Dawley (SD) rats were given orally by KMKKT at 250, 500, and 1000 mg/kg for 4 weeks. Mortality, clinical signs and measured change of body weight, food consumption and water consumption were observed. In addition, we performed ophthalmologic, urinary, hematological, blood serum biochemical and histopathological examination. Any general toxicity was not found in KMKKT treated group. Also, there were no significant differences in the parameters such as body weight, food consumption and water consumption, a lot of urine and blood factor levels except WBC, MCHC and Ca level compared with control group. Although WBC and MCHC were elevated in female rats and Ca level was decreased in male rats, these were within normal ranges. Finally, we determined that maximum tolerated dose (MTD) was 1000 mg/kg and no observed adverse effect level (NOAEL) was 500 mg/kg. Taken together, these results demonstrated that KMKKT is very safe to SD rats.

• 동의생리병리학회지
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• 제23권2호
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• pp.343-350
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• 2009

This study aimed to evaluate the anti-asthmatic effects of Samjajihwang-tang (SJT) using OVA-induced asthmatic mice model. Asthmatic mice model was conducted by repeated challenge of OVA using C57BL/6 mice. Each group was treated with distilled water, SJT (400 mg/kg and 200 mg/kg) extract or cyclosporin A (10 mg/kg) for the later 8 weeks, Penh (plethysmography and enhanced pause), immune cells subpopulation, eotaxin, IL-5, TNF-${\alpha}$, Anti-OVA-lgE in BALF (bronchoalveolar lavage), and lung tissue was analyzed, No cytotoxicity of SJT was shown on hFCs (human fibroblast cells). Administration of SJT significantly decreased Penh levels comparing to control group. SJT treatment significantly ameliorated the increase of total cells number and eosinophil including of immune cell subpopulation of $CD3^+/CD69^+$, $CCR3^+$, $B220^+/CD22^+$, $B220^+/CD45^+$, and $B220^+/lgE^+$ cells in BALF comparing to control group. Eotaxin, IL-5, TNF-${\alpha}$, and Anti-OVA-lgE level in BALF were significantly decreased by SJT treatment too. Histopathological finding verified the improvement of infiltration of inflammatory cells and collagen tissue in the SJT groups comparing to control group. These results strongly suggest that SJT would be a effective candidate for herbal-originated anti-asthmatic drug. However, this drug should be further studied for characterization of the accurate action and underlying mechanisms using variant disease model in the future.

• 대한화장품학회지
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• 제20권1호
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• pp.1-13
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• 1994

솔잎으로부터 프리 라디칼 소거 작용이 있는 물질을 분리하고, 여러 기기 분석 결과에 근거하여 4-hydroxy-5-methyl-3[2H]-furanone (HMF) 으로 동정하였다. 이 물질이 1, 1-diphenyl-2-picrylhydrazyl 프리 라디칼에 대한 소거 작용이 공지의 항산화 물질인 a-tocopherol, ascorbic acid와 유사하였다. HMF는 흰쥐 간 microsome분획에서 Fe(II)/ascorbate에 의해 유도된 지질 과산화를 억제하였으며, 배양 fibroblast 세포에서 자외선에 대한 보호효과를 나타내었다. 이 물질은 또한 tyrosine의 효소적 산화와 Dopa의 자동 산화를 억제하였을 뿐만 아니라 배양 murine melanoma 세포에서도 강력한 멜라닌 생성 억제 작용을 보였다. 피부 세포에서의 멜라닌 생성이 산화적 스트레스에 의해 유발되고 또 효소, 비효소적인 산화 반응을 통해 진행된다고 볼 때, HMF는 이러한 각 단계에서 항산화제로 작용하여 궁극적으로 세포에서의 멜라닌 생성을 막는 것으로 추론되었다.

• 대한화장품학회지
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• 제25권4호
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• pp.145-154
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• 1999

A novel retinol derivative, polyethoxylated retinamide(Medimin A) was synthesized, as an anti-aging agent. Collagen synthesis, skin permeation, stability, and toxicity of Medimin A were evaluated and compared with those of retinol and retinyl palmitate. In vitro collagen synthesis was evaluated by quantitative assay of $[^3H]-proline$ incorporation into collagenase sensitive protein in fibroblast cultures. For in vitro skin permeation experiments, Franz diffusion cells(effective diffusion area: 1,766 $cm^2$) and the excised skin of female hairless mouse aged 8 weeks were used, The stabilities of retinoids were evaluated at two different temperature($25^{\circ}C\;and\;40^{\circ}C$) and under UV in solubilized state and in O/W emulsion. To estimate the safety, acute oral toxicity, acute dermal toxicity, primary skin irritation, acute eye irritation and human patch test were performed. The effect of Medimin A on collagen synthesis was similar to that of retinol. The skin permeability of Medimin A was higher than those of retinol and retinyl palmitate. The Medimin A was more stable than retinol and retinyl palmitate. Medimin A was nontoxic in various toxicological tests. These results suggest that Medimin A would be a good anti-aging agent for enhancing bioavailability and stability.

• 대한화장품학회지
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• 제35권3호
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• pp.193-202
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• 2009

본 연구에서는 in vivo 광독성 시험을 대체할 수 있는 방법에 관한 연구를 수행하였다. 인체 유래의 섬유아세포를 활용하여 광독성 물질(promethazine, chlorpromazine chlortetracycline, 8-methoxypsoralen, neutral red, bithionol)과 비광독성 물질(cinnamic aldehyde, p-aminobenzoic acid, sodium lauryl sulfate, L-cysteine)을 평가하였다. 세포 생존율은 neutral red uptake (NRU)로 평가하였다. NRU 광독성 시험 결과 bithionol를 제외한 화합물에서 모두 in vivo 실험결과와 유사한 결과를 보였다. 같은 방법으로 화장품 성분인 $Medimin^{(R)}$ A, $Medimin^{(R)}$ D, $LG^{(R)}$ 106W, $Phytoclear^{(R)}$ EL-1, 단삼동 추출물, 미인초 추출물, 산거울 추출물, $Parsol^{(R)}$ MCX와 $Parsol^{(R)}$ 1789를 평가 하였다. 평가 결과 단삼동 추출물을 제외한 원료에서 광독성이 없는 것으로 평가되었다.

• 대한한방내과학회지
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• 제29권3호
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• pp.703-715
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• 2008

This experiments was performed to determine the effects of Gangzitongmaekeum(降脂通脈飮 : GTE) on antioxidation activity and hyperlipidemia induced by hypercholesterolemic diet in mice. The results obtained were as follows : 1. GTE showed a safety in cytotoxicity and toxicity of human fibroblast cells and liver. 2. GTE showed DPPH scavenging activity, superoxide dismutase (SOD)-like activity and inhibitory effect on reactive oxygen species (ROS). 3. GTE treated mice showed body and liver weight decrease, compared to the hyperlipidemia-induced control group. 4. GTE decreased total cholesterol and LDL cholesterol levels significantly, but HDL cholesterol levels not significantly. 5. GTE decreased triglyceride levels significantly. 6. Glucose levels in GTE treated mice significantly decreased compared to the hyperlipidemia-induced control group. 7. Albumin levels in GTE treated mice were similar with the hyperlipidemia-induced control group. 8. The lipophagy in liver compared with the control group tended to be decreased in GTE treated mice. In the change of aorta, the cell was regular and boundary of vessel wall was clear, compared to the hyperlipidemia-induced control group. 9. TBARS levels in GTE treated mice significantly decreased compared to the hyperlipidemia-induced control group. 10. The change of SOD and catalase activity significantly increased compared to the hyperlipidemia-induced control group. 11. ACAT mRNA level and HMG-CoA reductase mRNA levels in GTE treated mice significantly decreased compared to the hyperlipidemia-induced control group. Conclusion : These results suggest that Gangzitongmaekeum is effective in antioxidation activity and dietary hyperlipidemia-induced mice.

• 대한수의학회지
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• 제34권4호
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• pp.787-799
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• 1994

To elucidate morphologic lesion of porcine exudative epidermitis which is occurred sporadically in Korea, Staphylococcus hyicus subsp. hyicus isolated from the naturally affected pigs was inoculated to suckling pigs. The infected piglets were observed grossly and histopathologically. Although affected piglets were taking acute, subacute, or chronic course, some piglets suffered from chronic disease showed poor prognosis and marked growth depression. Affected peglets had erythematous skin on the face, ear, and abdomen and these localized lesions appear as brownish spots of exudative epidermitis and fromed crust in the early stage. But, after this stage, the skin were covered by viscous greasy exudate and formed blackish brown crust and appeared fissures and hypertrophy. Grossly, there has been hemorrhage with the removal of crust-like materials of epidermis and edematous subcutis. The superficial lymph nodes were edematous and swollen or congested and hemorrhagic. Some piglets had swollen ureters, cysts in the renal cortex, or polyarthritis. A few cases had mild edematous swelling of kidney, intestinal catarrh and congestion of brain. Microscopically, skin lesions had detachment of keralinized layer and parakeratosis of epidermis, hydropic degeneration of epidermal cell, and retrogressive degeneration of hair root sheath. Dermis had edema, and infiltration of neutrophils and mononuclear cells. As the disease was proceeded, there was marked perivasculitis with lots of mononuclear inflammatory cells. More chronic lesions formed granuloma-like bodies(nodules) due to more mononuclear, perivascular inflammatory cell infiltration and proliferation of fibroblast. Lots of plasma cells and eosinophils were also present in dermis. Epidermis was hyperplastic by proliferation of basal cells stratum germinativum and epidermal pegs often extended into the dermis. In secondary infection, lots of neutrophils could be seen in epidermis and derms. Kidney had neutrophilic infiltration, necrotic and cystic glomeruli, and dilation of renal tubules and ureters. Purulent arthritis was sometimes observed in joints. Three days old mice administrated Staphylococcus hyicus subsp hyicus subcutaneously before had focal congestion and hemorrhage, necrosis, and subcutaneous edema of the skin. This observation was also seen in the study of mice administrated exfoliatin toxin of Staphylococcus which evoked human staphylococcal scalded skin syndrome.

• Asian-Australasian Journal of Animal Sciences
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• 제27권12호
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• pp.1671-1677
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• 2014

Ras homolog enriched in brain (Rheb) and FK506 binding protein 38 (FKBP38) are two important regulatory proteins in the mammalian target of rapamycin (mTOR) pathway. There are contradictory data on the interaction between Rheb and FKBP38 in human cells, but this association has not been examined in cashmere goat cells. To investigate the interaction between Rheb and FKBP38, we overexpressed goat Rheb and FKBP38 in goat fetal fibroblasts, extracted whole proteins, and performed coimmunoprecipitation to detect them by western blot. We found Rheb binds directly to FKBP38. Then, we constructed bait vectors (pGBKT7-Rheb/FKBP38) and prey vectors (pGADT7-Rheb/FKBP38), and examined their interaction by yeast two-hybrid assay. Their direct interaction was observed, regardless of which plasmid served as the prey or bait vector. These results indicate that the 2 proteins interact directly in vivo. Novel evidence is presented on the mTOR signal pathway in Cashmere goat cells.

• 대한암한의학회지
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• 제13권1호
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• pp.13-24
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• 2008

Objective: To evaluate the effects of Bikihaun (BKH) on angiogenesis. Method: We examined the anti-angiogenic effect of BKH in invasion assay model. We performed proliferation assay, migration assay, tube formation assay and Chicken Chorioallantoic Membrane (CAM) assay. Results: In proliferation assay, at lower dose under 125 ${\mu}g/m{\ell}$ anti-angiogenesis effect of the group treated BKH made no difference with the control group. But, at the dose of 250 ${\mu}g/m{\ell}$ or more anti-angiogenesis effect of the group treated BKH showed more effective as compared to the control group. In migration assay, BKH did not affect migration of vascular endothelial cell. In tube formation assay, at lower dose under 100 ${\mu}g/m{\ell}$ showed mild effect of anti-tube formation. But, at the dose of 1000 ${\mu}g/m{\ell}$ showed more effective anti-tube formation. In CAM assay, BKH showed anti-angiogenesis effect at the dose of 10 ${\mu}g/m{\ell}$. Conclusion: BKH has antiangiogenetic properties in vitro.