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      • 고콜레스테롤 섭취가 흰쥐의 심근발달 및 혈액성분에 미치는 영향

        김용진(Yong Jin Kim),황호영(Ho Young Hwang),윤경아(Kyung Ah Yun) 한국체육교육학회 2000 한국체육교육학회지 Vol.4 No.2

        The present study was to elucidate the effect of endurance training and dietary cholesterol on myocardium development and blood constituent in rats. For the experiment male rats were used from the 4th week after their birth. The total of 128 Sprague-Dawley male rats were divided into 4 groups. the normal diet control group (NDCG), the normal diet training group (NDTG), the cholesterol diet control group (CDCG), and the cholesterol diet training group (GDTG). On each 4th, 8th, 12th, and 16th experimental week, eight rats of each group were sacrificed for tests. We examined the changes of serum enzymes by blood chemical analysis and the number of blood capillary in left ventricle among the experimental groups by morphometrical measurment. The results were as follows; In the normal diet training group (NDTG), it showed a decrease in weight compared with control group (NDCG) and was increased change of serum enzyme level, thickness of left ventricle wall, density of muscle fiber and capillary. In the cholesterol diet control group (CDCG), it was increase weight, thickness of left ventricle wall, density of muscle fiber and capillary and serum enzyme level with heart disease compared with training group (CDTG). `Therefore, it showed untrained and high cholesterol diet causes the change of myocardium in the development rat. These results suggest the endurance training have a preventing effect on the heart disease in hypercholesterol diet.

      • 고삼 추출물을 이용한 해독물질의 개발

        양희태,정승일,이영철,윤경아,백승화 한국식품영양학회 2002 韓國食品營養學會誌 Vol.15 No.1

        고삼 메탄올 추출물로부터 분리한 flavonoids를 IR, NMR등의 분광학적인 방법으로 Leachianone A(LA)과 Sophoraflavanone G(SFG)로 동정하였고, 카드뮴의 세포독성효과를 고삼 및 고삼추출물이 방어할 수 있는 지를 검색하기 위하여 NIG 3T3 세포에 고삼 및 LA와 SFG, LA+SFG를 처리하고 MTT assay 및 광학현미경으로 세포의 생존률을 검색하여 다음과 같은 결론은 얻었다. MIT의 흡광도는 카드뮴의 농도에 의존하여 감소하였으며, IC_50인 MTT50은 12.5??이었으며 카드뮴을 IC_50농도로 처리하고 고삼 및 LA, SFG, LA+SFG를 각 화합물의 자체 세포독성을 갖지 않는 농도로 처리한 후 MTT assay로 세포 생존율을 측정한 결과 LA, SFG, LA+SFG 처리군들은 카드뮴 처리군(MTT_50)에 비해 세포생존률이 증가되었으며 이들은 고삼 및 고삼추출물의 농도에 의존적으로 증가되었고 각각의 단일 화합물보다 두 화합물 LA+SFG 병용처리군에서 수복 효과가 더 좋은 것으로 나타났으며 광학현미경적 소견에서도 세포재생이 뚜렷하게 보였다. 이상과 같이 고삼에서 분리한 flavonoids가 카드뮴 독성에 의하여 손상된 NIH 3T3 섬유아세포의 재생효과에 영향이 있는 것으로 판단된다. The aim of this study was to assess the antitoxic compounds, flavanones (Leachinanone A=LA and Sophora flavanone G=SFG), from Sophora flavescents (S. Flavescens). We investigated the possibility of antitoxicity of LA and SFG against NIH 3T3 fibroblasts cell lines using colorimetric MTT[3-(4-5-dimethylthiazol-2-yl)-2-5diphenyl-2H-tetra-azolium bromide] assay. The results were as follows : After cadmium was treated against NIH 3T3 cell lines, we determined IC_50. Accordingly we have examined the detoxification effects of S. flavescens, LA and SFG under cadmium IC_50=12.5?? and was carried out to observe morphological changes by the light microscopic study. In NIH 3T3 cells, Sophora flavescens, LA, SFG and LA+ SFG showed inhibitory effects on the cytotoxicity of cadmium and these detoxication effects increased in proportion to the concentration of these drugs. These results suggest that LA and SFG from S. flavescens retain a potential antitoxic activity.

      • 12-O-Tetradecanoylphorbol-13-Acetate에 의한 HL-60 세포 분화유도중 Vimentin 유전자 전사조절에 대한 AP-1의 역할

        임규,김진희,권도원,김승민,이명선,윤경아,손미영,박종일,윤완희,황병두 충남대학교 생물공학연구소 1999 생물공학연구지 Vol.7 No.-

        Purpose: To gain insight on the role of AP-1 in transcriptional regulation of vimentin gene during differentiation of HL-60 cells by 12-O-tetradecanoylphorbol-13-acetate (TPA), the levels of vimentin mRNA and AP-1 have been investigated with Northern blot hybridization and DNA mobility shift assay. Materials and Methods: HL-60 cells were grown in RPMI 1640 medium supplemented with 10% heat-inactivated fetal calf serum and antibiotics in a humidified 5% CO_2 at 37℃. Total RNA was prepared by a modification of the method of Karlinsey et al. Northern blot hybridization was performed by the method of Virca et al. EcoRI fragment of pVIM-GEM was used as probe for vimentin mRNA. DNA mobility shift assay was performed by the method of Lim et al. End labeled DNA probe(Upper strand, 5'-CGCTTGATGAGTCAGCCG- 3') for AP-1 binding activity was mixed with nuclear extracts in a 20 μl reaction volume containing 300 mM KCI, 60 mM HEPES, pH 7.9, 25 mM MgCI_2, 1 mM EDTA, 1 mM DTT, 60% glycerol, and 2 ㎍ of poly[dI-dC]. Results: TPA increased vimentin mRNA levels, with maximal stimulation reached at 24 hr. The level of vimentin mRNA was induced in proportion to the concentration of TPA. TPA-induced vimentin mRNA was almost reduced by actinomycin-D pretreatment. TPA-induced stimulation of vimentin gene was completely reduced by staurosporin pretreatment. In DNA mobility shift assay, AP-1 newly appeared at 24 hr during TPA-induced differentiation and was almost not detected after the pretreatment of staurosporin. Conclusions: These results suggest that the induction of vimentin mRNA during TPA-dependent differentiation in HL-60 cells may be mediated by protein kinases C signal transduction and AP-1 is important to transcriptional regulation.

      • 12-O-Tetradecanoylphorbol-13-Acetate에 의한 HL-60 세포 분화유도중 Vimentin 유전자 전사조절에 대한 AP-1의 역할

        임규,김진희,권도원,김승민,이명선,윤경아,손미영,박종일,윤완희,황병두 忠南大學校 癌共同硏究所 1998 癌共同硏究所 硏究誌 Vol.2 No.1

        Purpose: To gain insight on the role of AP-1 in transcriptional regulation of vimentin gene during differentiation of HL-60 cells by 12-O-tetradecanoylphorbol-13-acetate (TPA), the levels of vimentin mRNA and AP-1 have been investigated with Northern blot hybridization and DNA mobility shift assay. Materials and Methods: HL-60 cells were grown in RPMI 1640 medium supplemented with 10% heat-inactivated fetal calf serum and antibiotics in a humidified 5% CO_(2) at 37°C. Total RNA was prepared by a modification of the method of Karlinsey et al. Northern blot hybridization was performed by the method of Virca et al. EcoRI fragment of pVIM-GEM was used as probe for vimentin mRNA. DNA mobility shift assay was performed by the method of Lim et al. End labeled DNA probe(Upper strand, 5'-CGCTTGATGAGTCAGCCG- 3') for AP-1 binding activity was mixed with nuclear extracts in a 20 μl reaction volume containing 300 mM KC1, 60 mM HEPES, pH 7.9, 25 mM MgCl_(2), 1 mM EDTA, 1 mM DTT, 60% glycerol, and 2μg of poly[dI-dC]. Results: TPA increased vimentin mRNA levels, with maximal stimulation reached at 24 hr. The level of vimentin mRNA was induced in proportion to the concentration of TPA. TPA-induced vimentin mRNA was almost reduced by actinomycin-D pretreatment. TPA-induced stimulation of vimentin gene was completely reduced by staurosporin pretreatment. In DNA mobility shift assay, AP-1 newly appeared at 24 hr during TPA-induced differentiation and was almost not detected after the pretreatment of staurosporin. Conclusions: These results suggest that the induction of vimentin mRNA during TPA-dependent differentiation in HL-60 cells may be mediated by protein kinases C signal transduction and AP-1 is important to transcriptional regulation.

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