• 사상체질의학회지
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• 제8권2호
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• pp.219-238
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• 1996

Effects of Taeyeumjoweetang on the obestity of mouse induced by gold thioglucose. It is researched to elucidate the effects of Taeyeumjoweetang on the obesity of mouse induced by gold thioglucose and the differentiation and growth of preadipocyte, 3T3-L1 cell. The result were as follows. 1. Taeyeumjoweetang extract improved the blood level of transaminase changed by the obesity of mouse induced by gold thioglucose. 2. Taeyeumjoweetang extract inhibited the increase of liver fat and body fat induced by the obesity of mouse induced by gold thioglucose. 3. Taeyeumjoweetang extract inhibited the increase of body weight induced by the obesity of mouse induced by gold thioglucose. 4. Taeyeumjoweetang extract inhibited the growth of undifferentiate preadipocyte 3T3-L1. 5. Taeyeumjoweetang extract showed the effects of inhibition on the differentiation of preadipocyte 3T3-L1. The above results suggested that the Taeyeumjoweetang extract may be used on the obesity induced by the overgrowth and differentiation of adipocyte, and the accumulation of fat in liver and body.

• 환경위생공학
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• 제22권1호
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• pp.75-83
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• 2007

Clove extract by methanol increased expression of the tyrosinase gene on B16 mouse melanoma cells containing tyrosinase promoter. $10{\mu}g/mL$ and $100{\mu}g/mL$ of the extract showed expression rate of the tyrosinase gene about 138% and 245%, respectively, compared with control. At $500{\mu}g/mL$, expression rate of the extract was impossible to measurement by high cytotoxicity. The solvent fraction of methylene chloride also exhibited highly expression rate as methanol extract. However, the solvent fractions of butyl alcohol and water showed repressive effect on expression of tyrosinase gene at $500{\mu}g/mL$. In MTT assay, cell survival rate of the extract exhibited similar to expression rate of tyrosinase gene. That is, $10{\mu}g/mL$ and $100{\mu}g/mL$ of the extract showed the cell survival rate about 128% and 187%, respectively.

• 대한의생명과학회지
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• 제9권3호
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• pp.111-121
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• 2003

Recombinant single chain Fv (scFv) antibodies offer many advantages over mouse monoclonal antibodies such as faster clearance from blood, improved tumor localization, reduced human anti-mouse antibody (HAMA) response, and the availability to manipulate the scFv through genetic approaches. The recombinant phage display was constructed using lym-l hybridoma cells as a source of genetic starting material. mRNA was isolated from the corresponding antibodies hybridoma cells. VH and VL cDNA were amplified with RT-PCR and linked with ScFv by linker DNA to form ScFv DNA, which then were inserted into phagemid pCANTAB5E. The phage of positive clones selected with tube containing raji lymphoma cell and infected by competent E. coli HB2151 to express soluble scFv. The scFv lym-l was secreted into the cytosol and culture supernatant and shown to be of expected size (approximately 32 kDa) by western blot. An active scFv lym-l could be produced in E. coli with soluble form and high yield from hybridoma cell line, using phage display system. Immunoreactivity indicated that scFv lym1 showed a potential biding affinity against the raji lymphoma cell as its parental antibody (intact lym-l Ab).

• IMMUNE NETWORK
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• 제8권4호
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• pp.137-142
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• 2008

Background: CD11c, also known as integrin alpha x, is one of the optimum markers of dendritic cells. However, the regulation of the CD11c expression in mouse has not been identified yet. In this study, in order to analyze the regulation of CD11c expression, the promoter of CD11c was cloned and characterized. Methods: To identify the promoter portion, various sizes of what are considered to be CD11c promoter fragments was amplified by polymerase chain reaction (PCR), using mouse genomic DNA as a template. After sequence was obtained, these fragments were transfected into various cell lines including mouse dendritic cell lines such as JAWSII and DC2.4 and L929 as control cell line.. The promoter activity of three promoter fragments was measured and compared by luciferase activity in the transfected cells. Results: Three clones with size of 1kb, 3kb and 6kb were obtained from mouse genomic DNA. Flow cytometry analysis of JAWSII cells revealed that 52% of the cells expressed CD11c, which was confirmed by RT-PCR analysis. On the contrary, L929 and DC 2.4 cells did not express CD11c. The CD11c+ JAWSII cells were enriched from 52% to 90% with cell sorter. The comparative luciferase activity analyisis demonstrated that the region responsible for tissue specific expression was contained within -3 kb and the clone with size of 3 kb particularly showed higher luciferase activity than 6 kb and 1 kb clones. Conclusion: The CD11c promoter region containing the region responsible for tissue specificity was successfully cloned and -3 kb region showed the highest activity.

• 환경생물
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• 제26권1호
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• pp.30-35
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• 2008

곰피추출물의 CCD-986sk cell line monolayer (human fibroblast, KCBL-21947)에 대한 피부세포 생리활성효과를 측정하고, 또한 곰피추출물의 Clone M-3 mouse melano-cyte cell line에 대한 melanin formation 저해효과를 측정하기 위해 in vitro레벨에서 실험을 실시하였다. 곰피는 다년생 갈조류의 일종으로 이 종은 한국 연안해역에서 중요한 1차생산자의 역할을 담당하고 있는 종이며 연안 생태계에 있어서 생태학적으로나 수산업상 매우 중요한 위치를 점유하고 있다. 최근 이러한 대형 갈조류에 관한 연구는 주로 생태학적인 관점에서 이루어져 왔으며(Notoya and Aruga 1990), 유주자의 발아와 핵분열(Yabu and Notoya 1985) 등에 관한 연구가 주로 일본의 연구자들에 의하여 보고된 바 있고, 국내에서는 Park et al. (1994)이 부산만 인근해역 곰피의 생장과 연령 조성에 관한 연구를 수행하였고, 순간접착제를 이용한 곰피의 이식 등에 관한 연구(최 등 2002)가 보고된 바 있다. 그러나 지금까지 곰피의 식품 또는 약품제제로서의 기능성을 밝히는 연구는 아직 미미한 실정으로 이에 대한 체계적인 연구가 부족한 실정이다. 따라서 본 연구는 곰피추출물에 풍부하게 함유된 fucoidan, L-Ascorbic 산 및 collagen 등에 주목하여 곰피의 피부노화방지제로서의 가능성을 체계적으로 연구하기 위한 일환으로 우선 곰피추출물이 피부재생, 주름살제거 및 피부미백작용에 효과를 나타내는지 확인하기 위해 CCD-986sk cell line 및 Clone M-3 mouse melanocyte cell line을 이용하여 in vitro 수준에서 피부세포 생리활성 효과와 melanin formation 저해효과를 측정하였다. 그 결과 CCD-986sk cell line에 대한 세포생리활성 효과 실험에서 시험군인 곰피추출물 투여군은 대조군에 비해 강한 세포생리활성 효과를 나타내어 유의성 있는 실험결과를 보여주었고, Clone M-3 mouse melanocyte cell에 대한 melanin pigment 저해활성 효과 실험에서도 곰피추출물은 대조군에 비해 melanin pigment 생합성을 유의성 있게 감소시키는 실험결과를 나타내어 매우 흥미로운 결과를 보여주었다. 여러 가지 보고에 의하면 그동안 미국, 영국, 일본 등 구미 선진국의 과학자들은 intrinsic problems (피부구조 및 생리기능의 노쇠)과 extrinsic problems (자외선노출 및 스트레스)을 skin aging의 근본원인으로 규정짓고 피부 콜라겐과 엘라스틴을 재생시켜 피부 horny layer의 주름을 제거시키는 수 많은 종류의 chemical drugs를 개발하였고 현재도 개발 중에 있다 (Lavker et al. 1987; Choi and Oh 1997; Han et al. 1998). 그러나 이런 chemical에 비해 생체에 대한 부작용이나 세포독성이 상대적으로 적은 천연물을 이용하여 anti-skin aging drug을 개발하는 연구는 그 동안 부족했던 것이 현실이다. 한국은 오랜 역사를 통하여 화학물질이 아닌 다수의 천연물들을 임상에 적용하여온 역사를 가지고 있고 따라서 본 연구팀은 우리 남해안의 청정해역에서 자라는 곰피의 항피부 노화방지제로서의 개발 가능성에 주목하여 본 연구를 진행하였다. 연구 결과, 곰피추출물은 피부노화 및 skindarkness에 대한 대체치료물질 (alternative therapeutic agent)로서의 충분한 가능성을 보여주었다.

• Toxicological Research
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• 제14권2호
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• pp.171-176
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• 1998

The biological effects of the water extracts of Rhus Verniciflua Stokes (RVS) were evaluated by protection against hydroxyl radicals. Antioxidative activities were measured using both 1,1-diphenyl-2-picrylhydrazyl (DPPH) and thiocyanate method. Also we used the Glucose oxidase (GO) 20 mU/$\textrm{m}{\ell}$ hydroxyl radical generating system in mouse forebrain cell culture. Water was used for ex-traction from RVS as a solvent which has high polarity especially. In DPPH method, the antioxidative activities of the crude water extract were stronger than any other extracts of low polar-solvents. In the antioxidative effects of mouse forebrain culture using 20 mU/$\textrm{m}{\ell}$ GO, cell viabilities were evaluated 65.6%, 68.8% at 1 $\mu\textrm{g}$. 5 $\mu\textrm{g}$ addition of crude water extracts (30 mg/$\textrm{m}{\ell}$) respectively. 10 $\mu\textrm{g}$ addition of crude water extracts had more than 86.1% cell viabilities, P<0.0l significantly, compared with the group treated with GO alone. In comparison with the antioxidative activities of several commercial antioxidants (ascorbic acid, $\alpha$-tocopherol, catalase, serum), 273 $\mu\textrm{g}$/$\textrm{m}{\ell}$ addition of crude water extracts (300 $\mu\textrm{g}$/$\textrm{m}{\ell}$) showed equivalent antioxidative effect to 25 uM ascorbic acid.

• 한국식품영양학회지
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• 제13권5호
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• pp.411-418
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• 2000

결명자차, 녹차 및 밤차가 인체의 생체기능활성화에 미치는 영향을 알아보기 위하여 국소뇌혈류량 및 혈압, in vitro상에서의 면역세포의 활성화, 그리고 암세포가 이식된 동물에서의 면역세포 활성화와 암세포의 증식억제 효과를 관찰한 결과 다음과 같은 결론을 얻었다. 1. 단당류는 현미녹차와 결명자차에는 glucose, ga-lactose등이 들어 있으나 밤차에는 glucose, gal-actose, mannose가 들어 있다. 2. 아미노산은 현미녹차, 밤차, 결명자차 순으로 들어 있다. 3. 카페인은 현미녹차에는 들어 있으나 밤차와 결명자차에는 들어 있지 않았다. 4. 결명자 차는 국소뇌혈류량을 증가시킨 반면 밤차는 감소시켰다. 5. 녹차는 혈압을 증가시켰다. 6. 밤차는 in vitro 상에서 흉선세포와 비장세포의 증식을 유의성있게 증가시켰다. 7. 결명자차와 녹차는 in bitro 상에서 흉선세포의 증식을 감소시켰다. 8. 결명자차와 녹차는 L1210세포가 이식된 동물의 흉선세포 증식을 감소시켰다. 9. 밤차는 L1210세포가 이식된 동물의 비장세포 증식을 촉진시켰다. 10. 밤차와 결명자차는 L1210세포가 이식된 동물의 암세포 증식을 억제하였다.

• Biomolecules & Therapeutics
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• 제14권3호
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• pp.143-147
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• 2006

In the mouse leukemic monocyte cell line RAW 264.7, the vacuolar-type $(H^+)$-ATPase (V-ATPase) inhibitor bafilomycin $A_1$ at 10 and 100 nM decreased cell growth and survival as determined by 3-(4,5-dimethyl(thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in a concentration-dependent manner. At such concentrations, bafilomycin $A_1$ induced nitric oxide (NO) production through the expression of inducible nitric oxide synthase (iNOS). The bafilomycin $A_1$-induced NO production was inhibited by the NOS inhibitor $N^G$-monomethyl-L-arginine acetate (L-NMMA). Our findings suggest that the V-ATPase inhibitor bafilomycin $A_1$ induces NO production through the expression of iNOS protein.

• 생명과학회지
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• 제22권1호
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• pp.49-54
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• 2012

이소플라본 genistein이 마우스 전지방 3T3-L1세포주의 성장과 지방세포형성과정에 미치는 영향을 연구하였다. 그 결과, 마우스 전지방 3T3-L1 세포주의 성장이 처리한 genistein 농도 의존적으로 저해되었다. 또한, Oil Red O 염색에 의하여 50 ${\mu}M$ genistein 처리에 의해 지방세포형성과정이 억제됨을 확인하였다. 이러한 genistein에 의한 지방세포성장 억제효과의 분자생물학적 기전을 연구하기 위하여, oligo DNA 마이크로어레이 실험을 수행하였다. 발현이 증가된 유전자 중 항비만, 항염증 활성과 관련이 있는 것으로 알려져 있는 sirtuin-1 유전자를 선택하여 발현변화를 확인하였다. 처리한 네종류의 파이토케미칼(resveratrol, capsaicin, daidzein, genistein)에 의해 sirtuin-1의 발현이 증가됨을 확인하였고, 이 중 genistein에 의한 sirtuin-1 유전자의 발현이 가장 높았다. 또한, sirtuin-1-siRNA에 의한 sirtuin-1 유전자의 발현 감소에 의해 지방세포형성 억제과정이 복구됨을 확인하였다. 이러한 연구결과는, genistein에 의한 전지방세포의 성장 저해와 지방세포형성의 저해과정을 이해하는데 도움을 줄 것으로 기대된다.

• 대한의생명과학회지
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• 제10권3호
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• pp.211-217
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• 2004

For discrimination of ductal and ascinar cells, we isolated a single-chain variable domain fragment (scFv) antibody against ductal cells of salivary gland using phage display technique. From the spleen of a mouse immunized with ductal cell lysate, total RNA was prepared and used as a template for cDNA synthesis of antibody genes. The scFv genes were constructed with variable domain genes of heavy and light chain and were introduced into pCANTAB5E to construct phage scFv library. The phage particles specific for acinar cells were screened by subtraction using immunotubes coated with acinar and ductal cell lysate and enzyme-linked immunoabsorbance assay (ELISA). The characteristics of the scFv were determined by immunohistochemistry (IHC) and the result indicated that the isolated scFv has the specificity against ductal cells of salivary glands and tubules of kidney. And the scFv has an unique binding activity specific for Hashimoto's thyroiditis. The nucleotide sequence of isolated scFv gene was determined and revealed that V/sub H/ belongs to the mouse H-chain family subgroup IB and V/sub L/ to the mouse L-chain family subgroup III.