• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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• pp.261-261
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• 1994

Bovine milk xanthine oxidase (E.C.1.1.3.22, XO) purchased from Sigma Chemical Co. had the three protein fragments below 150 kDa on 7.5％ SDS-PAGE, which did not show enzyme activity. To remove these fragments, the enzyme preparation was further purified through Sephadex G-200 column chromatography. Two peaks exhibiting enzymatic activity were separated very closely to the void volume, which were revealed as two different enzyme forms, dimeric and monomeric, confirmed by activity staining on native PAGE. Anti sera-against each of the two enzyme forms were raised by subcutaneous injection at multiple sites on the back of rabbits during 4 weeks. On the immunodiffusion test, it was found that both of the antisera of the two forms could react with each other, which implied that their epitopes were identical In the Western blot analysis of mouse liver cytosol fraction, it was found that rabbit anti-XO antibody bound well with the protein band of monomeric mouse liver XO of about 150kDa. Based on this result, mouse liver cDNA 1 ibrary was screened by in situ hybridizat ion wi th rabbi t anti -XO antibody as probe. Through the immunological screening, recombinant phages giving positive signal by the production of XO were selected and further purified. To validate these clones, purified phages were lysogenized in E. coli Y1089 and their lysates were analysed for enzyme activity and immunoreactivity, It was verified that lysates of the purified recombinant phage lysogens exhibited the enzymatic activity as well as bound wi th XO antibody, when induced by IPTG. The above results assert that selected recombinant phage carries mouse liver XO gene.

• Natural Product Sciences
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• 제19권2호
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• pp.173-178
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• 2013

The protective effect of SAPP, an extract from a novel herbal complex, on acute liver injury was investigated using mouse animal model in this study. The content of total phenol in SAPP was increased at dose dependent manner. Consistent with the content of total phenol, SAPP showed the significant anti-oxidative effects on 1, 1-diphenyl-2-picrylhydrazyl (DPPH) method. Acute liver injury was induced by D-galactosamine (D-GalN) in mouse. Treatment with SAPP significantly reduced the level of alanine transaminase (ALT) and aspartate transaminase (AST) in serum. Histological observation revealed that whereas D-GalN treated mouse showed vacuolization of hepatocytes, sinusoidal dilation and congestion, loss of cell boundaries and ballooning degeneration, loss of architecture and cell necrosis, treatment with SAPP improved D-GalN-induced liver injury. These results suggest that SAPP shows protective effects against D-GalN-induced hepatotoxicity in vivo acute mouse model.

• Reproductive and Developmental Biology
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• 제35권4호
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• pp.427-434
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• 2011

Mitochondria diseases have been reported to involve structural and functional defects of complex I-V. Especially, many of these diseases are known to be related to dysfunction of mitochondrial proton-translocating NADH-ubiquinone oxidoreductase (complex I). The dysfunction of mitochondria complex I is associated with neurodegenerative disorders, such as Parkinson's disease, Huntington's disease, and Leber's hereditary optic neuropathy (LHON). Mammalian mitochondrial proton-translocating NADH-quinone oxidoreductase (complex I) is largest and consists of at least 46 different subunits. In contrast, the NDI1 gene of Saccharomyces cerevisiae is a single subunit rotenone-insensitive NADH-quinone oxidoreductase that is located on the matrix side of the inner mitochondrial membrane. The Saccharomyces cerevisiae NDI1 gene using a recombinant adeno-associated virus vector (rAAV-NDI1) was successfully expressed in AML12 mouse liver hepatocytes and the NDI1-transduced cells were able to grow in media containing rotenone. In contrast, control cells that did not receive the NDI1 gene failed to survive. The expressed Ndi1 enzyme was recognized to be localized in mitochondria by confocal immunofluorescence microscopic analyses and immunoblotting. Using digitonin-permeabilized cells, it was shown that the NADH oxidase activity of the NDI1-transduced cells was not affected by rotenone which is inhibitor of complex I, but was inhibited by antimycin A. Furthermore, these results indicate that Ndi1 can be functionally expressed in the AML12 mouse liver hepatocytes. It is conceivable that the NDI1 gene is powerful tool for gene therapy of mitochondrial diseases caused by complex I deficiency. In the future, we will attempt to functionally express the NDI1 gene in mouse embryonic stem (mES) cell.

• Reproductive and Developmental Biology
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• 제31권3호
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• pp.145-152
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• 2007

본 연구에서는 SV40 Tag을 마우스 albumin 유전자의 promoter/enhancer 조절 하에 발현하도록 설계된 재조합 유전자를 마우스 1세포기 수정란에 미세 주입하여 형질 전환마우스를 제작하고 이들의 인체 간암 모델로써의 적합성을 조사하였다. 형질 전환이 확인된 총 11개체의 founder 생쥐들 중 4개체가 간암을 일으켰고, 두 개체는 신장암을, 한 개체는 피부 및 뇌에서 종양을 각각 일으켰다. 이들로부터 외래 유전자를 계대 유전하는 3가계를 얻었다(#1-2, #1-6, #1-11). 이들 가계의 자소들에서 8주령(#1-2, #1-6)혹은 10주령(#1-11)시부터 간암이 반복적으로 발생되었으며, #1-11 founder개체에서 폐로 암세포가 전이된 것 외에는 다른 조직에서의 형태학적 변이가 발견되지 않았다. 간암 발생은 조직학적 변화에 따라 3단계로 나눌 수 있었다. 즉, 출생에서 3주령까지는 간세포의 과량증식을 보이나 세포핵의 이상은 관찰되지 않았으며, 4주령부터 7주령(#1-2, #1-6) 혹은 9주령(#1-11)까지는 diffuse liver cell dysplasia를 나타내지만 tumor nodule은 발견되지 않았고, 그 이후에는 liver dysplasia를 배경으로 간암이 발생하였다. 본 연구에서 작출한 간암 모델 마우스는 인체 간암과 일부 유사한 소견을 보였는바 인체 간암 기전 연구를 위한 유용한 동물 모델로 이용할 수 있을 것으로 생각된다.

• 한국식품과학회지
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• 제30권3호
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• pp.693-696
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• 1998

Chitin의 탈아세틸화로 얻어지는 화합물인 chitosan과 chitooligosaccharide는 다양한 생리활성 기능을 나타내므로 생물의학적인 응용에 많은 관심이 집중되고 있다. 본 연구는 수용성 chitooligosaccharide가 생쥐의 간 기능에 미치는 영향을 조사하기 위하여 수행되었다. 6주 동안 3% cholesterol를 식이한 생쥐에서는 혈장 cholesterol 함량이 증가하였으나, 1% chitooligosaccharide와 3% cholesterol을 함께 식이한 생쥐에서는 혈장 cholesterol 함량이 23% 정도 감소하였다. 그러나 혈장내 HDL-cholesterol과 간내 cholesterol 함량에는 유의한 차이가 없었다. 한편, 15% ethanol을 8주간 투여한 생쥐에서는 inflammation, necrosis, 지방구의 축적, GPT의 활성 증가 등 간 상해가 관찰되었다. 그와 반면에 0.5% chitooligosaccharide와 15% ethanol을 함께 투여한 생쥐에서는 ethanol에 의해 유발된 간 상해가 현저히 억제되어 지방구의 축적이 관찰되지 않았고, 혈장의 GPT 활성이 25%정도 감소하였다. 따라서, chitooligosaccharide가 생쥐에서 혈장 cholesterol 함량을 감소시키고, 알코올성 지방간을 예방하는 등 부분적인 기능이 있음을 보여주었다.

• 생명과학회지
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• 제11권2호
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• pp.111-120
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• 2001

The pathological aspects of purified ricin from the seeds of the castor oil plant, Ricinus communis, were examined, using light and transmission electron microscopy. ICR mice were exposed to ricin by peritoneal injection with 100 ng/1 $m\ell$ PBS(pH 7.0) mouse and histological observations on the liver, spleen, thymus, lung and heart were carried out at intervals up to 48 h after exposures. All the organs examined were damaged by ricin. Among the organs, the spleen and thymus; immune organs were the most sensitive to ricin, whereas the effect delayed in the liver, lung and heart. Furthermore, the immune cells in each organ were the most sensitive to ricin. Accordingly, the effect of ricin on the organs seems to be affected by the immune cells existed in each organ, In each organ, the immune cells showed apoptotic changes, while the capillary endothelial and parenchymal cells showed necrotic changes.

• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5405-5408
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• 2012

Curcumin (CM), a biphenyl compound, possesses anti-inflammatory, antioxidant and antimicrobial activity. MicroRNAs (miRNAs) are small noncoding RNAs which regulate gene expression and the molecular mechanisms of several biological processes. Liver fibrosis is a major cause of hepatic dysfunction and cancer and there are few effective therapies emphasizing the need for new approaches to control. The present study was conducted to investigate the effect of curcumin (CM) on liver fibrosis through modulating the expression level of miRNAs (199 and 200), the main miRNAs associated with liver fibrosis. Induction of liver fibrosis by carbon tetrachloride ($CCL_4$) was confirmed by histopathological examination. Mice were divided into 3 groups: group 1 were i.p injected with 10% $CCL_4$ twice weekly for 4 weeks and then once a week for the next 4 weeks followed by 4 weeks with olive oil only. Group 2 were i.p injected with 10% $CCL_4$ twice weekly for 4 weeks and then once a week for the next 4 weeks followed by curcumin (5 mg/mouse/day) once daily for the next 4 weeks. The third group was injected with olive oil. The expression level of miR-199 and miR-200 and some of their targeted genes were measured by real time PCR. miRNA (199 and 200) levels were significantly elevated in liver fibrotic tissues compared to control groups. Curcumin was significantly returned the expression levels of mir-199 and -200 with their associated target gene nearly to their normal levels. This is the first study that highlighted the effect of curcumin on liver fibrosis through regulation of miRNAs.

• Journal of Nutrition and Health
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• 제24권4호
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• pp.356-365
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• 1991

유전에 의해 비만증을 타고나는 비만쥐 (ob/ob mouse)의 간으로 부터 유리된 세포막과 핵의 $L-triiodothyronine(T_3)$ 결합 능력이 그들과 한 배에서 태어난 정상 체중의 쥐들의 것들과 비교되었다. 세포막이 hypotonic 용액과 discontinuous sucrose density를 사용하여 원심분리기로 분리되었으며, 세포 각 부분의 marker enzyme들의 activity로 세포막의 순도가 측정되었다. 핵은 $Triton{\times}100$를 사용하여 윈심분리기로 얻어졌다. $T_3$ 수용체의 Bmax(최대결합용량)와 Kd(dissociation constant)가 세포막 혹은 핵을 여러 농도의 $^{125}I-T_3$와 함께 일정시간 incubation 시킨 후, 그 binding이 reverse Scatchard analysis에 의하여 계산하여 얻어졌다. 모든 실험과정은 비만쥐와 정상쥐에 대하여 평행으로 진행되었다. 간 핵의 $T_3$ 수용체의 최대결함용량은 비만쥐가 정상 체증의 쥐 보다 유의적으로 적었으나(p<0.001), $T_3$에 대한 친화력에는 차이가 없었다. 이는 이전의 보고들의 결과를 확인해 주는 것이다. 세포막에 있는 $T_3$ 수용체의 최대결합 능력과 친화력은 비만쥐와 정상쥐 간에 유의적인 차이가 없는 것으로 밝혀졌다. 이는 비만쥐의 세포막에 있는 $T_3$수용체의 기능에는 결함이 없음을 나타내며, 비만쥐의 말초조직에서 손상된 갑상선 홀몬의 작용은 세포막 수용체에 결합한 이후에 일어나는 과정에 원인이 있다는 것을 의미하고, 따라서 핵에 있는 $T_3$수용체의 결함이 비만쥐 (ob/ob mouse)의 비만증의 근본적인 원인일 수 있다는 제안을 뒷받침하여 주고 있다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1995년도 춘계학술대회
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• pp.128-128
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• 1995

To investigate the mechanism of the regulation of cytochrome P450IA1 gene expression, ethoxyresorufin deethylase(EROD) and benzo(a)pyrene hydroxylase in B6 mouse liver, in isolated perfused rat liver system. and in B6 mouse hepatocyte Hepa-I cells were examined. In C57BL/6N mouse, 3-methylcholan- throne( 3MC ) treatment have resulted in the stimulation of EROD activity based on fluorometry by 2.79 fold comparirng with that of control. Measurement of mRNA of cytochrome P450 was carried out by either nothern blot or dot blot analysis. Findings are similar to that of studies with enzymes. Furhtermore, when RTPCR method was applied to detect mRNA in Hepa I cell and liver tissues the results were more clear. Cytochrome P450IA1 upstream DNA containing CAT construct was transfected into Hepa-1 cells. After transfection of CAT construct, 3MC and flavonoids, such as, chrysin, hesperetin, kaempferol, morin, myricetin and aminoyrine were treated. 48 Hours after treatments, cells were harvested and assayed for CAT mRNA by RTPCR. 3MC treatment to hepa I cells transfected with trout P450IA1-CAT construct increased CAT mRNA by 2.81 fold when it was compared with that of control. This increase CAT mRNA was decreased by concomitantly treated flavonoids and aminopyrine. The level of CAT protein was 29.2-58.0% of 3MC stimulated CAT protein. Results of this study suggested that RTPCR seems to be a very good method to study regulation of gene expression in liver tissue or Hepa cells.

• 한국환경보건학회지
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• 제22권1호
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• pp.51-56
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• 1996

In order to detect the T-2 toxin accumulation in the animal tissues, T-2 toxin, produced by Fusarium sporotrichioides M-1-1, was injected to mouse by 0, 1 and 2 mg per kilogram of body weight, respectively, and T-2 toxin extracted from serum and organs were analyzed by the indirected competitive ELISA. The indirect competitive ELISA established in the laboratory can be check less than 0.1 ppb level of T-2 toxin and average recovery of T-2 toxin spiked was 80~113% in animal samples such as serum, liver and kidney. After 6 weeks of treatment with 2 mg of T-2 toxin per kg body weight, T-2 toxin was accumulated in serum (133.0 ng/ml), liver(1.4 ng/g) and kidney(14.3 ng/g) of mouse injected with 2 mg of toxin per kg body weight.