• 대한한의학회지
• /
• 제23권3호
• /
• pp.33-42
• /
• 2002

Objective : This study was performed to investigate the activity of Whagan-Jeon (huaganjian) in protection against acetaminophen (AAP)-induced hepatotoxicity and the possible mechanisms in vivo. Methods : The following were performed : Serum ALT, depletion of hepatic glutathione (GSH) levels, the microsomal p. nitrophenol hydroxylation activity, microsomal aniline hydroxylation activity, genomic DNA fragmentation and its reversal, hepatic glutathione-S-transferase (GST) activity, and hepatic NAD(P)H:quinone oxidoreductase (QR) activity Results : Whagan-Jeon (huaganjian) protected against AAP-inducedhepatotoxicity by the increase of GSH levels, inhibition of P450 2E1-specific metabolic activities, attenuation of hepatic DNA damage, and induction of GST and QR activities in vivo. Conclusions : In conclusion, Whagan-Jeon (huaganjian) was effective in protection against AAP-induced hepatoxicity.

• 농약과학회지
• /
• 제3권3호
• /
• pp.27-36
• /
• 1999

쥐에 있어서 carbamate계 살충제 carbofuran의 독성에 미치는 phenobarbital sodium(PB) 또는 3-methylcholanthrene(3-MC)의 영향과 작용기작을 효소적 측면에서 구명할 목적으로 이들을 단독 또는 조합으로 경구투여 하여 in vivo 효소활성을 조사하였다. Acetylcholinesterase(AChE)와 butyrylcholinesterase(BuCheE)의 효소활성은 carbofuran 3.8 mg/kg을 투여하였을 때 48시간까지 $20{\sim}70%$ 범위의 저해를 보였고, carbofuran과 PB 또는 3-MC를 조합투여하였을 때 효소활성은 초기에 감소하다가 24시간 후에는 대조구와 비슷한 수준을 나타냈다. Glutathione S-transferase(GST)의 경우 carbofuran만을 투여하였을 때 초기($0.5{\sim}6$ hr)에 $15{\sim}35%$의 저해를 보였으나, carbofuran과 PB 또는 3-MC의 조합투여시 초기에는 약간 저해를 보이다가 3시간 후에는 대조군과 유사한 효소활성을 보였고, 6시간 후에는 대조군에 비해 활성이 20%이상 증가하였다. UDP-glucuronosyltransferase(UDPGI) 및 cytochrome P-450 효소계의 효소활성은 carbofuran과 PB 또는 3-MC를 조합 투여하였을 때 투여 후 6시간까지는 carbofuran만의 투여에 비해 효소활성이 $2.6{\sim}2.8$배 이상 높았다. 이상의 결과에서 PB 및 3-MC의 투여가 이들 효소활성을 유도하므로써 carbofuran의 독성으로부터 쥐를 보호한 것으로 판단된다.

• 생명과학회지
• /
• 제9권3호
• /
• pp.253-261
• /
• 1999

Leptin gene, an obesity gene, has been known to involve in the regulation of food intake and body weight. It is also thought to be related to the glucose metabolism, insulin secretion and type II diabetes mellitus. Recently, the production of recombinant leptin protein has been attempted for the application in the treatment of obesity and the correction of hereditary obesity and type II diabetes. In the present study, leptin cDNA was cloned from mouse fat cells by RT-PCR and prokaryotic expression of leptin was attempted in order ot prepare a leptin-specific antigen. Immunization of a rabbit with the leptin-specific antigen into a rabbit resulted in the generation of leptin-specific antiserum that could be useful in the detection of leption expressed in various tissues. The sequence of leptin cDNA prepared in the present study wa identical to the previously reported one. Transformation of E. coli(DH5a) cells with the leptin cDNA-inserted translation vector, pGEX-4T-3-leptin followed by treatment with IPTG (0.1mM) resulted in the expression of a large amount of GST-leptin fusion protein with a molecular weight of 44 KDa as an inclusion body. Denaturation of the insoluble fusion protein by 8M urea, 6M guanidium-HCI or 0.1% 2-mercaptoethanol followed by a slow oxidation could not solubilize the inclusion body. The cell extract was subjected to SDS-PAGE and GST-leptin protein electroeluted from the gel was then injected into a rabbit subcutaneously for the immunization. Anti-GST-leptin rabbit antiserum which had a cross reactivity to the GST-leptin protein was generated. Leptin protein expressed in mouse brain and fat tissues was detected by Western blot immunodetection system using the antiserum generated in the present study.

• 한국식품영양과학회지
• /
• 제38권2호
• /
• pp.154-159
• /
• 2009

알코올에 의해 유도된 간 손상에 대한 지구자 추출물의 보호효과를 연구하였다. HepG2/2E1 세포에서 알코올로 유도된 ROS 생성과 산화적 손상에 대한 지구자 추출물 보호효과를 확인하였다. C57BL/6마우스를 대조군(NC), 알코올군(ET), 알코올과 지구자 추출물 1 g/kg body weight 투여군(ET-HD)으로 나누었다. 5 g/kg body weight의 알코올을 1주일간 ET와 ET-HD군에 투여하였다. 알코올 투여는 혈청 alanine amintransferase(ALT), aspartate aminotransferase(AST) 및 alkaline phosphatase(ALP)를 증가시키고, 지구자 추출물은 이러한 간 기능 지표효소의 증가를 억제시켰다. 간조직의 항산화 효소 활성은 알코올 투여에 의해 감소되었고, ET-HD군에서 SOD 및 GST 활성은 ET군과 비교하여 통계적으로 유의하게 높아졌다. GSH 함량은 ET군에서 NC군에 비하여 유의적으로 낮아졌고, ET-HD군에서 ET군과 비교하여 통계적으로 유의하게 높아졌으며, NC군과 유사한 함량을 나타내어 간 보호 효과를 확인할 수 있었다. 지질과산화물 함량은 ET-HD군과 NC군이 유사한 함량을 나타냄으로써 알코올에 의해 유도된 지질과산화물 증가에 의한 간손상으로부터 지구자 추출물의 보호 효과를 보여 주었다. 이상의 결과로부터, 지구자 추출물은 세포 및 동물 모델에서 알코올로 유도된 간 손상으로부터 항산화 방어 대사의 증가와 지질과산화율의 감소에 의해 간세포 보호 활성을 나타냄을 확인하였다. 이에 지구자 추출물은 알코올성 간 손상으로부터 보호 효과를 갖는 소재로 활용될 수 있을 것으로 사료된다.

• Biomolecules & Therapeutics
• /
• 제8권2호
• /
• pp.153-159
• /
• 2000

A novel serine/threonine protein phosphatase with EF-hand motif, which belongs to PPEF family was partially cloned from rat brain cDNA by employing RT-PCR method. The size of the amplified clone was 1.6kbp. The amplified DNA was subcloned into pGEM-T-Easy vector and the resulting plasmid was maned as pGEM-rPPEF2. The nucleuotide sequence is shared by 88% with that of mouse PPEF-2 cDNA, and the deduced amino acid sequence reveal 92% homology with that of mouse PPEF-2 cDNA. The N-terminal region of the cloned rat brain PPEF contains a putative phosphatase catalytic domain (PP domain) and the C-terminal region contains multiple $Ca^{2+}$ binding sites (EF region). The putative catalytic domin (PP) and the EF-hand motif (EF) regions were subcloned into pGEX4T-1 and were overexpressed in E. coli DH5 as glutathione-S-transferase (GST) fusion proteins. Expression of the desired fusion protein was identified by SDS-PAGE and also by immunoblot analysis using monoclonal antibody against GST. The recombinant proteins were purified by glutathione-agarose chromatography. This report is first to demonstrate the cloning of PPEF family from rat brain tissues. The clone reported here would be invaluable for the investigation of the role of this new type-phosphatase in rat brain.

• 한국전기전자재료학회논문지
• /
• 제23권3호
• /
• pp.199-205
• /
• 2010

The crystallization speed (v) of amorphous (InTe)$_x$(GeTe) (x = 0.1, 0.3 and 0.5) films and their thermal, optical and electrical behaviors have been investigated using nano-pulse scanner (wavelength = 658 nm, laser beam diameter < 2 ${\mu}m$), X-ray diffraction (XRD), 4-point probe and UV-vis-IR spectrophotometer. These results were compared with those of $Ge_2Sb_2Te_5$ (GST) film, comprehensively utilized for phase-change random access memory (PRAM). Both v-value and thermal stability of (InTe)$_{0.1}$(GeTe) and (InTe)$_{0.3}$(GeTe) films could be enhanced in comparison with those of the GST. Contrarily, the v-value in the (InTe)$_{0.5}$(GeTe) film was so drastically deteriorated that we could not quantitatively evaluate it. This deterioration is thought because amorphous (InTe)$_{0.5}$(GeTe) film has relatively high reflectance, resulting in too low absorption to cause the crystallization. Conclusively, it could be thought that a proper compositional (InTe)$_x$(GeTe) films (e.g., x < 0.3) may be good candidates with both high crystallization speed and thermal stability for PRAM application.

• 동아시아식생활학회지
• /
• 제11권4호
• /
• pp.259-267
• /
• 2001

납중독에 따른 식이내 Se 함량이 간조직의 산화적 손상을 방어할 수 있는 가를 관찰하기 위해 140g 내외의 흰쥐 Sprague-Dawley종 수컷을 정상군과 식이 내에 납 함량을 2,000 ppm 투여한 납투여 실험군으로 나누고 다시 식이내 Se 수준에 따라 0 ppm(Pb0군), 0.5 ppm(PbS군), 1.0 ppm(PbSS군) 식이군으로 나누어 4주간 사육한 후 간장 중 SOD. GSH-Px, GST 등 항산화효소의 활성을 측정하고, GSH 및 비타민 E 함량을 측정하였다. 또한 전자현미경을 통하여 간세포의 소기관을 관찰하였다. 1. 간장 중 SOD는 Pb0군이 다른 군에 비해 다소 높았다. GSH-Px 활성은 Pb0군은 정상군에 비해 현저하게 감소되었으나 PbS군과 PbSS군간은 Pb0군에 비해 증가되었다. GST활성은 Pb0군만 정상군에 비해 감소되었고 PbS, PbSS,군은 정상군 수준이었다. 2. GSH 함량은 정상군에 비해 Pb0군에서 낮았으나 정상군과 PbS. PbSS군은 유의적인 차이가 없었다. GSSG는 GSH와 반대로 Pb0군에서 증가되었으며 GSH/GSSG는 GSH함량과 같은 경향이었다. 또한 간조직 중의 비타민 E함량은 Pb0군은 정상군에 비해 약 50% 정도의 수준이었고 PbSS군과 PbS군은 Pb0군에 비해 증가되었다. 3. 전자현미경적 관찰에서는 RER의 감소, 라이소솜의 증가, 미토콘드리아의 종창 등을 나타내었는데 그 정도는 PbSS, PbS, Pb0군 순으로 심하게 나타났다. 이상의 결과로 식이 중 Se의 다량 첨가는 납중독으로 인한 간조직의 항산화계를 강화시키고 세포 소기관들의 산화적 손상을 현저하게 완화시킬 수 있음을 알 수 있다.

• 한국응용약물학회:학술대회논문집
• /
• 한국응용약물학회 1997년도 춘계학술대회
• /
• pp.95-95
• /
• 1997

Among the various receptor molecules discovered so far the ${\beta}$2-adrenergic receptors have been regarded as excellent model systems for the so called 7 transmembrane helix receptor and have been the focus of extensive studies. For the analysis of receptor structure and function a monoclonal antibody plays a crucial role, thus providing useful tools for the study of receptor. However, because of the minute quantity of receptor molecules which could be obtained from natural sources, the generation of specific monoclonal antibody against receptor molecules from the purified receptors has been regarded as virtually impractical in consideration of cost and experimental times. The purpose of the present study was to generate and characterize a monoclonal antibody against human ${\beta}$2-adrenergic receptor. For the production of antibody, C-terminal regions of the human ${\beta}$2-adrenergic receptor was produced as a fusion protein with Glutathion S-transferase (GST) in E. Coli. The expression of the fusion protein was identified by SDS-PAGE and Western blot using monoclonal anti-GST antibody. The fusion protein was purified to an apparent homogeniety by affinity chromatography with Glutathion Sepharose CL-4B and used as an antigen for the immunization of BALB/C mice. The Production of monoclonal antibody was achieved by fusion of the immunized spleen cells and SP/2-0 myeloma cells. Positive hybridomas were screened by ELISA and were cloned by two consecutive rounds of limiting dilution. The monoclonal antibody produced in this study (mAb${\beta}$C02) was IgM type and purified by immunoaffinity chromatography using anti-mouse IgM agarose as an affinity matrix. MAb${\beta}$C02 showed strong and specific immunoreactivity against both the fusion protein and human ${\beta}$2-adrenergic receptor in ELISA and Western blot. The molecular weight of immunoreactive band was 64 kDa and exactly coincided with the previously reported molecular weight of ${\beta}$2-adrenergic recepters. The results of the present study suggest that mAb${\beta}$C02 may be used for the study of receptor function and regulation in normal or nonphysiological status.

• 한국식품영양과학회지
• /
• 제27권6호
• /
• pp.1262-1266
• /
• 1998

The purpose of this study was to investigate peroxidative damage and antioxidative defense systems such as superoxide dismutase(SOD), glutathione peroxidase(GSH Px), glutathione S transferase (GST) and vitamin E of liver in rat exposed microwave. Sprague Dawley male rats 200$\pm$10gm were randomly assigned to normal and microwave(MW) groups. After rats were irradiated with microwave at frequency of 2.45GHz for 15min, the change patterns of antioxidative defense system and peroxidative damage of liver tissue in MW group were investigated for 16 days(the 2nd, 4th, 6th, 8th and 16th days) compared with those of normal group. The activity of superoxide dismutase(SOD) in MW group was increased at the 2nd day compared with that of normal group, but not significantly. The glutathione peroxidase(GSH Px) in MW group was decreased to 24% and 25% at the 4th and 6th days, respectively, compoared with that of normal group, but GSH Px was increased to level of normal group at the 16th day. The activity of glutathione S transferase(GST) in MW group was decreased at the 2nd day after irradiated with microwave, but GST showed to that of normal group at the 16th day. The content of vitamin E in MW group was lower than that of normal group at the 6th and 8th days after the irradiation, but was recovered to the level of normal group at the 16th days. The content of thi obarbituric acid reactive substances(TBARS) of liver in MW group was increased to 28.9%, 53.8%, 69.7% and 30.2% of normal group at the 2nd, 4th, 6th and 8th days after the irradiation, respectively, but recovered to the level of normal group at the 16th day. The present results indicated that antiox idative defense systems of rats irradiated microwave was weaken more than that of normal group, which lead to acceleration of lipid peroxidation.

• BMB Reports
• /
• 제35권6호
• /
• pp.615-622
• /
• 2002

The purpose of this study was to determine the effects of dietary garlic powder on diethylnitrosamine (DEN)-induced hepatocarcinogenesis and cytochrome P450 (CYP) enzymes in weaning male Sprague-Dawley rats by using the medium-term bioassay system of Ito et al. The rats were fed diets that contained 0, 0.5, 2.0 or 5.0% garlic powder for 8 weeks, beginning the diets with the intraperitoneal (i.p.) injection of DEN. The areas of placental glutathione S-transferase (GST-P) positive foci, an effective marker for DEN-initiated lesions, were significantly decreased in the rats that were fed garlic-powder diets; the numbers were significantly decreased only in the 2.0 and 5.0% garlic-powder diets. The p-nitrophenol hydroxylase (PNPH) activities and protein levels of CYP 2E1 in the hepatic microsomes of the rats that were fed the 2.0 and 5.0% garlic powder diet were much lower than those of the basal-diet groups. Pentoxyresorufin O-dealkylase (PROD) activity and CYP 2B1 protein level were not influenced by the garlic-powder diets and carcinogen treatment. Therefore, the suppression of CYP 2E1 by garlic in the diet might influence the formation of preneoplastic foci during hepatocarcinogenesis in rats that are initiated with DEN.