• The Journal of Internal Korean Medicine
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• v.25 no.1
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• pp.81-91
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• 2004

Objectives : The aim of this study is to investigate the inhibitory effect of Chungganhaeju-Tang on alcohol induced human hepatic cell apoptosis by synthesis of glutathione. Methods : The amount of glutathione in HepG2 cell was measured with colorimetric glutathione assay kit and glutathione-conjugated CDNB(1-chloro-2,4-dinitrobenzene) at $37^{\circ}C$ and then measured by spectrometry to assess the activity of glutathione S-transferase. Results : The synthesis of glutathione and the activity of glutathione S-transferase in HepG2 cell were promoted by Chungganhaeju-Tang and increased in dose/time-dependent manner. Chungganhaeju-Tang inhibited apoptosis induced by ethanol and acetaldehyde dependent to treatment dosage. In Buthione sulfoximine, a glutathione synthesis inhibitor, treated case, the synthesis of glutathione was inhibited and in Chungganhaeju-Tang treated case, the synthesis of glutathione is promoted with or without Buthione sulfoximine. The present findings suggest that Chungganhaeju-Tang inhibits alcohol induced apoptosis by synthesis of glutathione in HepG2 cell. Conclusions : The result indicates that Chungganhaeju-Tang protects human hepatic cell by glutathione synthesis and made the liver recover from alcohol induced damage.

• Korean Journal of Environmental Agriculture
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• v.16 no.1
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• pp.37-43
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• 1997

Tolerance mechanism to simazine (6-chloro-N,N'-diethyl-1,3,5-triazine-2,4-diamine) in Coix lacryma-jobi was investigated with respect to herbicide detoxification via glutathione conjugation. Simazine was initially absorbed by seedlings of C. lacryma-jobi and corn, but after 12 hours of treatment, no significant difference in simazine absorption was found in both species. Simazine absorbed was rapidly metabolized to glutathione-simazine conjugate. One to six hours after treatment, metabolism was approximately 2-fold faster in C. lacryma-jobi than in corn. Glutathione content was found 1.5- and 2.3-fold higher in coleoptile and root of C. lacryma-jobi, respectively, compared with corn. In both species, the highest concentration of glutathione was found in coleoptile tissue. Glutathione S-transferase that exhibits activity with 1-chloro-2,4-dinitrobenzene was not significantly different between two species. However, glutathione S-transferase activity with simazine was approximately 2-fold greater in C. lacryma-jobi than in corn. The glutathione S-transferase activity was 20 to 30% greater in shoot of either species than in root. Fast protein liquid chromatography-anion exchange column was used to separate glutathione S-transferase isozymes in coleoptiles of C. lacryma-jobi and corn. A peak of glutathione S-transferase activity with 1-chloro-2,4-dinitrobenzene and two peaks of glutathione S-transferase activity with simazine from C. lacryma-jobi were coeluted with those from corn, but showed greater activity than in the case of corn. Another glutathione S-transferase isozyme that exhibits activity with simazine was detected in the elution of C. lacryma-jobi extract, but not in corn. Electron transport in chloroplast thylakoids isolated from leaves of both species was equally sensitive to simazine applied at 1 to 100 nM. These results indicate that the simazine tolerance in C. lacryma-jobi is due to its capacity to detoxify the herbicide via glutathione conjugation, which is positively correlated with the level of glutathione content and glutathione S-transferase activity.

• Journal of Plant Biotechnology
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• v.4 no.3
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• pp.117-122
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• 2002

Cotton Glutathione S-Transferase(GST: EC 2.5.1.18) was cloned and Gh-5 cDNA was overexpressed in tobacco (Nicotiana tabacum) plants. The transformation of cotton GST in tobacco plant was confirmed by northern blot analysis. Type I and Type II transcript patterns were identified in Gh-5 transgenic tobacco plants. Type I transcripts was only discussed in this paper. Glutathione and 1-chloro-2,4-dinitrobenzene (CDNB) were used as the substrates, and the activity of GST in the type I transgenic plants was about 2.5-fold higher than the non-expressers and wild type tobacco plants. The expression of cotton GST in tobacco plants proved that Gh-5 could be translated into functional protein. Type I transgenic plants produced functional GST in the cells. Type I showed higher GST specific activity than Type II in the transgenic plants. Control and transgenic seedlings were grown in the growth chamber and under the light at 15$^{\circ}C$, and the effects of cotton GST in the seedlings was evaluated. The growth rate of Gh-5 overexpressors was better than the control and non-transgenic tobacco plants. Salinity tolerance was also analyzed on the seeds of transgenic plants. Seeds of Gh-5 overexpressors and the wild type tobacco seedlings were germinated and grown at 0, 50, 100, 150, and 200 mM NaCl solution. Gh-5 transgenic seedlings showed higher growth rate over control seedlings at both 50 and 100 mM NaCl solution. But at 0, 150, and 200 mM NaCl concentration, the difference in growth rate was not detected.