• Journal of Life Science
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• v.9 no.3
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• pp.253-261
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• 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.

• The Journal of Internal Korean Medicine
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• v.28 no.3
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• pp.453-463
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• 2007

Backgrounds : Glutathione-s-transferase (GST) is a kind of phase II metabolism enzyme and plays an important role in the detoxification of various toxic chemicals. It was reported that the genetic polymorphism of GSTM1 and GSTT1 genes may be responsible for asthma development and susceptibility to allergy. Traditional oriental medicine uses a unique diagnostic technique. differentiation-syndrome. to analyze signs and symptoms of patients synthetically. Through differentiation-syndrome. asthma patients can be divided into two groups: the deficiency syndrome group (DSG) and the excess syndrome group (ESG). Objectives : The purpose of this study was to investigate the possible association of GST gene polymorphism with clinical phenotype by differentiation-syndrome of bronchial asthma patients. Materials and Methods : One hundred and ten participants were evaluated by pulmonary function test. Patients with 53 DSG and 31 ESG by differentiation-syndrome were assessed for genetic analysis. GSTM1 and GSTT1 deletion polymorphism was performed by polymerase chain reaction (PCR). Results : GSTM1 gene deletion was detected in 43.4% of individuals in the DSG and in 38.71 % in the ESG. The distribution of GSTM1 polymorphism between DSG and ESG was not significantly different [$x^2$=0.1767, p=0.6742; OR(95% CI)=1.2139(0.4915-2.9979)]. The proportion of GSTT1 null genotypes was 41.51% in the DGS and 45.16% in the ESG. The distribution of GSTT1 polymorphism between DSG and ESG was also not significantly different [$x^2$=0.1065, p=0.7442; OR(95% CI)=0.8618(0.3525-2.1065)]. In the combined analysis of GSTM1 and GSTT1 genes, the frequency of both null type of GSTM1/GSTT1 genes was not significantly different from both positive type of GSTM1/GSTT1 genes[$x^2$=0.0768, p=0.7817; OR(95% CI)=1.2000(0.3303-4.3602)] Conclusions : These results indicate that polymorphism of the GST gene might not be associated with the symptomatic classification of DSG and ESG by differentiation-syndrome in Korean asthmatics.

• Korean Journal of Community Nutrition
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• v.2 no.5
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• pp.735-744
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• 1997

The effect of green tea drinking on the hepatocellular chemical cacinogenesis have been studied. Placental glutathione S-transferase(GST-P) positive foci area in a liver tissue, contents of thiobarbituric acid reactive substances(TBARS), total cytochrome P450 and glucose 6-phospphatase(G6P) activity in hepatic microsomes were investigated. Weaning Sprague-Dawley male rats were fed AIN-76A diet with deionized water or green tea infusion, Rats of CTR and CTR+ groups were provided deionized water while GTI and GTI+ groups were provided green tea instead of deionized water for the entire experimental period of 13weeks. Rats of GTP and GTP + groups had deionized water for the first 6 weeks and switched to green tea for the last 7weeks of the experimental period. CTR+, GTI +, and GTP + groups were carcinogen treated groups, Diethylnitrosamine(DEN) was injected as a single dose of 200mg/kg body weight intraperitoneally after 4 weeks of feeding. 2-Acetyla-minofluorene(AAF) was used as a carcinogen proliferater and suppled in the diets of carcinogen treated rats as 0.02% content for the last 6weeks starting from 2weeks after DEN injection. Rats were sacrificed after 13week weeks of feeding. The area and number of GST-P positive foci detected in carcinogen treated rats were decreased by green tea ingestion but when timing and duration of green tea ingestion was delayed after promotion period as in GTP + group, GST-P positive foci were not decreased as much as in GTI+ group. TBARS contents of carcinogen treated rats decreased by 13weeks of green tea ingestion but GTP groups did not show statiscally significant differences. G6P activities tended to decrease by carcinogen treatment but changes were not statiscally significant by green tea ingestion. Total cytochrome P450 contents were increased by carcinogen treatment. Thirteen weeks of green tea ingestion (GTI) also increased to total cytochrome P450 contents while 7weeks of green tea ingestion(GTP) did show any effects. These results suggest that green tea has suppressive effects on hepatocellular chemical carcinogenesis probably through the activities of antioxidant compounds. (Korean J Community utrition 2(5) : 735∼744, 1997)

• The Korean Journal of Pesticide Science
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• v.9 no.4
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• pp.338-346
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• 2005

Serum alanine aminotransferase(ALT), aspartate aminotransferase (AST), hepatic glutathione, glutathione S-transferase(GST), cytochrome P450 and cytochrome P450 reductase activity were measured to investigate the effects of hepatic detoxication system and metabolic activities of carbendazim in Sprague Dawley(S.D.) male rat at dose levels of 375, 750 or 1,500 mg/kg body weight. Serum alanine aminotransferase(ALT) and aspartate aminotransferase(AST) activities were slightly increased in all test groups after 120 minutes of administration. Glutathione was increased about 20% at high and medium dose level within 120 minutes after administration, while activity of glutathione S-transferase was decreased $36{\sim}50%$. However, the enzyme activity was recovered from all test groups after 240 minutes of administration. Cytochrome P450 and activity of cytochrome P450 reductase were decreased $25{\sim}50%$ until 120 minutes after administration, but recovered after 240 minutes.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.11
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• pp.6475-6479
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• 2013

Objectives: Glutathione S-transferase (GST) isoenzymes play important roles in resistance to cell apoptosis and carcinogenesis. We aimed to establish the relationship between GST expression and the prognosis of upper urinary tract urothelial carcinoma (UTT-UC) in Taiwan. Methods: This study retrospectively reviewed 46 patients with pathologically confirmed UUT-UC at Kaohsiung Medical University Hospital. In each patient, expression of GSTT1 and GSTP1 was compared between urothelial carcinoma and normal urothelial cells by Western blotting. Results: GSTP1 expression in the UUT-UC cells was significantly higher than that in normal urothelial cells (1.6 fold, p<0.001). Expression of GSTT1 was significantly associated with the invasiveness of the carcinoma (p=0.006). Conclusions: In UUT-UC, GSTP1 might be a potential tumor marker, whereas high GSTT1 expression could be used as an indicator of cancer progression. This study is the first to demonstrate potential applications of different GST isoenzymes for biomolecular analysis of UUT-UCs in Taiwan.

• Archives of Pharmacal Research
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• v.28 no.3
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• pp.311-318
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• 2005

Oxidative stress has been reported to elevate ceramide level during cell death. The purpose of the present study was to modulate cell death in relation to cellular glutathione (GSH) level and GST (glutathione S-transferase) expression by regulating the sphingolipid metabolism. LLC­PK1 cells were treated with H$_2$O$_2$ in the absence of serum to induce cell death. Subsequent to exposure to H$_2$O$_2$, LLC-PK1 cells were treated with desipramine, sphingomyelinase inhibitor, and N-acetylcysteine (NAC), GSH substrate. Based on comparative visual observation with H202-treated control cells, it was observed that 0.5 $\mu$M of desipramine and 25 $\mu$M of NAC exhibited about 90 and $95\%$ of cytoprotection, respectively, against H$_2$O$_2$-induced cell death. Desipramine and NAC lowered the release of LDH activity by 36 and $3\%$ respectively, when compared to $71\%$ in H$_2$O$_2$-exposed cells. Cellular glutathione level in 500 $\mu$M H202-treated cells was reduced to 890 pmol as compared to control level of 1198 pmol per mg protein. GST P1-1 expression was decreased in H$_2$O$_2$-treated cells compared to healthy normal cells. In conclusion, it has been inferred that H$_2$O$_2$-induced cell death is closely related to cellular GSH level and GST P1-1 expression in LLC-PK1 cells and occurs via ceramide elevation by sphingomyelinase activation.

• BMB Reports
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• v.33 no.6
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• pp.469-475
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• 2000

The purpose of this study was to examine the chemopreventive potential of taurine at various levels on the diethylnitrosamine (DEN)·induced hepatocarcinogenesis. Male Sprague-Dawley rats were fed on diets containing 0, 1, 2, 3% taurine or 5% ${\beta}-alanine$ for taurine depletion. Then they were treated with DEN and 2/3 partial hepatectomy. The number of placental glutathione S-transferase positive ($GST-P^+$) foci, as a preneoplastic marker in the 1 % taurine group was lower than the control diet group. However the difference was insignificant. Although taurine diets reduced the thiobarbituric acid reactive substance (TBARS) level, the number of $GST-P^+$ foci was increased in 3% taurine diet group. The 1 % taurine diet increased the glutathione (GSH) level and GST activity, however they unfortunately did not suppress the foci formation. In the 3% taurine group, the GSH level and GSH peroxidase (GPx) activity were significantly decreased. Excess taurine supplementation of the pharmaceutical dose worked against hepatic chemoprevention, which might result from modulation of GPx activity and GSH utility. On the contrary, taurine might work as an antioxidant against TBARS production as the 1 % taurine diet increased GSH level. The potency of the cancer preventive effect of taurine still remains and further studies should investigate the effect of taurine with less than 1 % levels on the prevention of hepatic cancer.

• Environmental Mutagens and Carcinogens
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• v.11 no.2
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• pp.118-133
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• 1991

신생랫드를 이용한 다장기 발암모델의 전암단계 병변에서 간장의 GST-P 활성도와 발암 과정에 영향을 미치는 phenobarbital (PB)의 발암촉진효과 및 병리조직학적 소견을 관찰하였다. 신생랫드를 150마리 3군으로 나누어 diethylnitrosamine (DEN : 100mg/kg,i,p.), N-me-thyl-N-nitrosourea (MNU : 20mg/kg,i,p.), N-bis(2-hydroxypropyl)nirtosamine(DHPN : 0.1% D. W)을 각각 0, 3, 6 주에 투여하였다. 또한 PB (0.5% in basal diet)를 7주부터 계속 투여하여 8, 12, 20주에 경시적인 부검을 실시하였다.

• Korean Journal of Poultry Science
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• v.37 no.1
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• pp.23-33
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• 2010

The current study was performed to develop natural bio-active substances as additives for the production of high quality broiler chickens. A total of 120 male 3 day-old broiler chicks were randomly allocated to CON (control), GK2.5 (ginkgo leaf 2.5%), GK5.0 (ginkgo leaf 5.0%), PK2.5 (pumpkin 2.5%) and PK5.0 (pumpkin 5.0%) of five groups in cages (24 birds per group). All birds were fed corresponding diets from 3 to 35 d of age and determined growth performance and biological parameters including blood biochemical profiles, antioxidant status and intestinal microflora. During the entire feeding trial, GK5.0 and PK5.0 groups resulted in a significantly (P<0.05) higher FCR than GK2.5 and PK2.5 groups. Plasma triglyceride significantly (P<0.05) increased in GK5.0 group compared with the other groups, and the level of alanine transaminase (ALT) increased (P<0.05) in GK5.0 and PK5.0 groups compared with that in PK2.5 group. Dietary addition of ginkgo leaf and pumpkin significantly (P<0.05) increased superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the small intestine. Also, the addition of 2.5% ginkgo leaf significantly (P<0.05) increased the activities of SOD, GSH-Px and glutathione-S-transferase (GST) in the liver. Futhermore, muscle GST activity significantly (P<0.05) enhanced by dietary addition of ginko leaf and pumpkin. However, the level of lipid peroxidation (MDA) in the small intestine and muscle turned to be higher (P<0.05) in PK5.0 group. The colony forming units (CFU) of E. coli in intestinal digesta significantly (P<0.05) decreased in both ginko leaf and pumpkin supplemented groups compared with CON group. In conclusion, dietary addition of 2.5% ginko leaf and pumpkin as dietary sources can be applicable for the production of high quality broiler chickens.

• Proceedings of the KSCN Conference
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• 1998.05a
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• pp.78-78
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• 1998