꽈리 추출물 투여가 마우스 간 Glutathione S-transferase 활성에 미치는 영향

Lee, Sang-il 啓明專門大學 産業開發硏究所 1999 啓明硏究論叢 Vol.17 No.2

천연물로부터 생리활성 물질을 검색하는 일환으로 우리나라에 널리 분포하는 꽈리를 대상으로 하여 여러단계를 거쳐 n-butanol 추출물을 얻고 실험동물에 용량별 및 기간별로 투여하면서 간조직 glutathione S-transferase의 활성 및 glutathione의 함량에 미치는 영향을 검토하여 다음과 같은 성적을 얻었다. 꽈리 n-butanol 추출물을 4일간 체중 kg당 100, 200 및 400mg씩 복강내로 투여하였을 때, 체중당 간무게 및 혈청 alanine aminotransferase의 활성은 유의한 변동이 없었고 과산화지질의 함량은 투여량에 반비례하여 감소하였다. 한편 간조직 glutathione S-transferase의 환성은 꽈리 추출물 투여량과 투여기간에 반비례하여 억제되었으며, glutathione의 함량 변동도 이와 유사하였다. 한편 동력학적인 측면에서 glutathione S-transferase의 활성 변동을 관찰하였을 때, 꽈리 n-butanol 추출물을 투여한 실험군이 생리식염수를 주사한 대조군에 비해 Vmax치가 현저히 감소하였다. To evaluate the effect of ground cherry n-butanol extract on the hepatic glutathione S-transferse, 100, 200 or 400mg/kg of ground cherry n-butanol extract were daily given to mice. The ratio of liver weight per body weight and serum alanine aminotransferase activity were not significantly changed, whereas hepatic lipid peroxide content was decreased as dose dependent manner. And ground cherry n-butanol extract treatment to the mice led to decreased activity of glutathione S-transferase activity and decreased content of reduced glutathione compared with the only physiological saline treated mice. The glutathione S-transferase activity was decreased as time-dependent manner by the injection of ground cherry n-butanol extract (200mg/kg) for one, two or four days. And the Vmax value without affecting the Km value for 1-chloro-2,4-dinitrobenzene was decreased by the ground cherry n-butanol extract treatment. From the above result, it is concluded that the ground cherry n-butanol extract may decrease the hepatic glutathione S-transferase activity due to an alteration in the quantity of enzyme protein, but further research in this field is needed.

Candida sp. 변이주에 의한 Glutathione 생산

김대선,유재홍,윤성식,신원철 한국산업미생물학회 1993 한국미생물·생명공학회지 Vol.21 No.5

Glutathione 생산균주를 돌연변이시켜 변이주의 생산 최적조건 및 glutathione의 분리, 정제를 행하였다. Glutathione 생산을 위한 Candida sp. mutant의 배지조성은 glucose 1.5%, yeast extract 4.0%, KH_2PO_4 0.04%, biotin 5 ㎍/㎖ 및 L-cysteine 0.04%이었으며 온도는 25℃, pH 6.0에서 48시간 배양하였을 때 glutathione 생산이 가장 좋았다. Glutathione 생산을 위한 최적배지에서 Candida sp. mutant 균주에 의한 glutathione 생산량은 175 ㎍/㎖로 원균주보다 1.9배 정도 증가하였다. 표준 glutathione과 정제된 glutathione의 TLC, UV 및 IR spectrum을 비교하여 본 결과 동일한 구조를 가지고 있는 것으로 판단되었다. c For the overproduction of glutathione, Candida sp. mutant was isolated by the treatment with U.V. light. The highest glutathione production of Candida sp. mutant was obtained after shaking culture for 48 hours in the culture medium containing glucose 1.5%(W/V). yeast extract 4.0%(W/V), KH_2PO_4 0.04%(W/V), biotin 5 ㎍/㎖, and L-cysteine 0.04%(W/V). The optimal pH and temperature for the glutathione production were pH 6.0 and 25℃, respectively. The glutathione production of Candida sp. mutant under the optimal culture condition was 175 ㎍/㎖. The purified glutathione was identified to be the same as the authentic glutathione by TLC chromatogram, UV and IR spectrum.

A colorimetric and fluorescent probe for rapid detection of glutathione and its application to tissue specific bio-imaging in living cells and zebrafish

Chen, Liyan,Park, Jong-Su,Wu, Di,Kim, Cheol-Hee,Yoon, Juyoung Elsevier 2018 Sensors and actuators. B Chemical Vol.262 No.-

<P><B>Abstract</B></P> <P>Owing to the many physiological functions of glutathione, fluorescent sensing systems that selectively and sensitively detect this biothiol in biological systems are in great demand. In this study, we developed a rhodamine-based probe to detect glutathione. The probe, which contains an α-bromoamide reactive center, responds rapidly and highly selectively to glutathione in both a colorimetric and “off/on” fluorescence manner. Upon addition of glutathione to a solution of the probe, up to an 80-fold enhancement in the intensity of fluorescence at 579 nm takes place, which can be visually observed by using a hand-held UV lamp (365 nm). Moreover, following addition of glutathione to the solution of the probe undergoes a color change from colorless to pink in association with the development of an absorption band at 560 nm. Consequently, glutathione detection can be made using “naked-eyes”. The results of studies aimed at exploring the cell permeability and intracellular GSH detection ability of the probe show that the probe can be utilized to sense endogenous and exogenous glutathione in HeLa cells. Moreover, the probe can be utilized to interrogate the oxidation states of glutathione, which are crucial to the maintenance of intracellular redox balances. Either addition of hydrogen peroxide or a lipopolysaccharide to solutions containing the probe and glutathione results in a remarkable decrease in the intensity of fluorescence due to their metabolism is in conjunction with glutathione oxidation. More interestingly, the probe can be used to detect glutathione in zebrafish with a high specificity for olfactory pit tissue.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A rhodamine-based fluorescence probe for sensing glutathione was developed. </LI> <LI> Remarkable fluorescence enhancement and colorimetric change were achieved upon the addition of glutathione. </LI> <LI> The probe can be utilized to sense endogenous and exogenous glutathione in HeLa cells. </LI> <LI> The probe can be utilized to interrogate the oxidation states of glutathione. </LI> <LI> The probe can be utilized to detect glutathione in zebrafish with a high specificity for olfactory pit tissue. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Bromate가 흰쥐의 장기 Glutathione 함량에 미치는 영향

김나영,강혜옥,이무강,최종원 한국생명과학회 2003 생명과학회지 Vol.13 No.5

백서에 bromate의 장기간 섭취로 간 및 신장의 glutathion량이 감소되는데, 간과 신장은 유사한 감소양상을 나타냈고, 폐 및 혈액에서는 감소하는 경향은 있었으나 통계적인 유의성은 없었다. Bromate 섭취로$\gamma-glutamyl$-cysteine synthetase와 $\gamma-glutamyl$ transpeptidase 활성이 감소되었다. 따라서 간과 신장의 glutathione 감소로 $\gamma-glutamyl$cysteine synthetase 활성이 감소됨으로서 glutathione 합성저하에 의해 나타난 결과로 생각되고, 폐에서 $\gamma-glutamyl$cysteine synthetase 및 $\gamma-glutamyl$- transpep-tidase 활성에는 별다른 영향이 없었다. 혈액에서는$\gamma-glutamyl$cysteine synthetase와 $\gamma-glutamyl$ transpeptidase 활성 감소로 glutathione의 혈액내로 유입과 타장기로 유출이 모두 저하되어 glutathione량의 변화가 없는 것으로 생각된다. Bromate에 의한 장기내 glutathione량 감소는 유리기 소거기능이 저하되어 bromate에 의해서 생성된 유리기 제거가 미흡할 것으로 생각되므로 bromate 독성의 한 요인이 될 것으로 추측된다. The effects of bromate administration on glutathione were studied in rats. The contents of glutathione in the liver and kidney were significantly decreased but the alteration was not significant in lung and blood by bromate adminstration. The decrease occurred without concomitant increases in oxidized glutathione (GSSG) or in the GSSG/GSH+GSSG ratio. The activities of $\gamma-glutamyl$ cysteine synthetase in the liver and kidney were decreased by bromate administration. $\gamma-Glutamyl$ transpeptidase activities was significantly decreased in the kidney and not significantly decreased in the lung of bromate treated-rats. These results suggest that the decreased synthesis of glutathione by bromate may be an important reason for the decreased level of glutathione in the liver and kidney, thus the decreased glutathione transport would be a factor on the changes of glutathione contents in bromate-treated rats.

청간해주탕(淸肝解酒湯)이 인체간세포의 Glutathione 생성에 미치는 영향

윤여광,이장훈,우홍정,김영철,Yoon Yeo-Kwang,Lee Jang-Hoon,Woo Hong-Jung,Kim Young-Chul 대한한방내과학회 2004 大韓韓方內科學會誌 Vol.25 No.1

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.

항암제 내성 L1210세포의 Glutathione 대사 관련효소 유전자의 발현 양상

김성용,김정희,김재룡 영남대학교 의과대학 1995 Yeungnam University Journal of Medicine Vol.12 No.1

생쥐의 백혈청세포 L1210과 항암제에 대하여 내성이 유도된 L1210AdR, L1210VcR과 L1210Cis에서 glutathione의 농도와 glutathione의 합성 조절에 관여하는 -glutamylcysteine synthetase(GCS)와 -glutamyl transpeptidase(GGT), 세포 이물질을 축합하는데 촉매하는 glutathione S-trasferase(GST)의 효소 활성도와 유전자의 발현 여부를 관찰하였다. 세포내 glutathione농도(μM/㎎ protein)는 L1210이 0.41±0.003, L1210AdR가 0.73±0.006, L1210VcR은 1.16±0.060, L1210Cis가 2.19±0.282으로 모세포에 비하여 내성세포에서 통계적으로 유의한 증가를 관찰하였다. Buthionine sulfoxamine(BSO)를 1??농도로 첨가하여 12시간 배양한 세포들에서의 glutathione농도는 L1210이 88%, L1210AdR가 85%, L1210VcR이 89%, 그리고 L1210Cis는 79%의 감소를 보였다. GCS의 활성도(nM/㎎ protein/min)는 L1210이 104인데 비하여 L1210AdR가 128, L1210VcR는 227, 및 L1210Cis는 212로 증가하였다. GGT의 활성도(nM/㎎ protein/min)는 L1210이 2.15±0.531이었고, L1210AdR은 2.80±0.498, L1210VcR은 2.42±0.389, 그리고 L1210Cis는 2.98±0.623으로 내성인 세포들에서 증가하였으며 L1210AdR과 L1210Cis에서 유의하였다. GST활성도(nM/㎎ protein/min)는 L1210이 16.70±4.798이었고, L1210AdR은 14.51±3.402, L1210VcR과 L1210Cis가 약간의 증가를 보였으며, L1210AdR은 오히려 감소를 보였다. DNA의 slot blot에서 GCS, GGT, GST 유전자의 모세포와 내성세포간에 별다른 차이를 보이지 않았다. Northern hybridization에서 GCS는 약 4.5kb 크기의 band, GST-π는 약 1.05kb 크기의 band를 보였으며 내성세포 모두에서 발현 증가가 관찰되었다. GGT의 경우 크기가 다른 6개의 band가 보였으며 특히 11.5kb 크기의 band에서 L1210AdR과 L1210VcR의 발현이 증가하였으며, L1210VcR에서는 L1210과 다른 내성세포에서 보이는 1.95kb 크기의 band가 보이지 않고 2.2kb 크기의 다른 band가 관찰되었다. 이상에서 L1210AdR과 L1210VcR의 내성에는 mdr1 유전자가 관여하고, L1210Cis의 내성에는 특히 glutathione이 중요하다. GCS, GGT 및 GST등의 활성도 및 유전자의 발현도 내성세포들에서 증가하였으며 이중 GCS는 내성세포내의 glutathione 합성에 가장 중요한 조절인자라 할 수 있다. Glutathione(GSH) has a very important role in detoxification of cells and is closely related to antitumor drug-resistance of cancer cells. In order to evaluate the importance of glutathione metabolism in the drug-resistant cancer cells, the concentration of cellular GSH and activities of -glutamylcysteine synthetase(GCS), -glutamyl transpeptidase(GGT) and glutathione S-transferases(GST) in the adriamycin, vincristine, or cisplatin resistant L1210(L1210AdR, L1210VcR, or L1210Cis) sublines were measured. Expression and amplification of GCS, GGT, and GST-π genes were also observed in the parent L1210 and the drug-resistant L1210 sublines. The concentration of GSH was increased 5.34 fold in L1210Cis, 2.83 fold in L1210VcR, and 1.78 fold in L1210AdR, compared to L1210. The activities of GCS and GGT were increased in drug-resistant L1210 sublines. The GST activity was increased in L1210VcR and L1210Cis but decreased in L1210AdR compared to L1210. Expression of GCS, GGT, and GST-π genes were increased in the resistant L1210 sublines compare to the parent L1210 in northern blot analyses. Overexpression of GCS, GGT, and GST-π were observed in the resistant sublines, and the increases of the concentration of glutathione and the activities of GCS and GGT in the resistant sublines may be involved in a part of the drug-resistance in the resistant sublines.

노화와 간경변이 인체의 적혈구, 혈장 및 위 점막의 Glutathione 농도와 Glutathione Peroxidase 활성도에 대해 미치는 영향

백광호,김종혁,백일현,한태호,박현주,박상훈,서중산,박충기,유재영 대한노인병학회 2001 대한노인병학회지 Vol.5 No.3

가자(Terminalia chebula) 추출물이 흰쥐의 간장 활성에 미치는 영향

박종옥,이인섭,최종원 한국생명과학회 2004 생명과학회지 Vol.14 No.1

가자 추출물을 흰쥐에 일정량 투여한 후 간장에 미치는 영향을 관찰할 목적으로 여러 가지 효소량의 변동을 검토하였다. 간 손상의 지표로 사용되는 AST, ALT활성을 측정한 결과 AST, ALT모두에서 정상군보다 300 mg/kg의 용량으로 2주 시료 투여한 군의 함량이 2배 이상 증가되는 것으로 나타났다. 가자 추출물이 지질과산화물 생성에 미치는 효과를 관찰한 결과, 300 mg/kg의 용량으로 2주 시료 투여한 군함량이 정상군에 비해 135.43%, 173.9% 정도 증가하는 결과를 나타내었다. 간 조직의 지질과산화물 반응에 관여하는 것으로 알려진 XO, AO, AD 및 AH활성에 대한 효과를 관찰한 결과 마이크로솜 분획에 존재하는 AD, AH의 활성은 저해 효과가 없었고, 세포질 효소인 XO, AO의 활성은 300 mg/kg의 용량으로 2주 시료 투여한 군 함량이 정상군에 비해 약 2배 증가됨을 볼 수 있었다. 가자 추출물에 의한 간 조직중 glutathione농도에 미치는 영향에 대해 관찰한 결과 2주 동안 300 mg/kg의 용량으로 투여한 군의 glutathione농도는 정상군에 비해 약 75% 감소하는 것으로 나타났다 가자 추출물 투여 후 glutathione의 함량 감소를 경감시키는 기전을 알아볼 목적으로 glutathione 합성에 관여하는 $\gamma$-GCS의 활성과 산화형 glutathione을 환원형 glutathione으로 환원시키는 GR의 활성을 관찰한 결과 정상군에 비해 GR이 56.6%, $\gamma$-GCS가 6.7% 정도 감소하는 것으로 나타났다. 이러한 glutathione의 함량 변동은 산화형 glutathione을 환원형 glutathione으로 환원시키는 GR의 활성에 영향을 주어 나타나는 결과로 생각된다. 가자 추출물이 지질과산화의 해독계에 미치는 영향을 관찰할 목적으로 catalase, GP 및 SOD의 활성을 측정한 결과 catalase, GP의 활성은 각각 77.5%, 64.3% 감소하는 결과를 나타냈으며, SOD의 활성은 정상군에 비해 약 3배 증가하는 것으로 나타났다. In this study, we investigated the effect of Teminalia Chebula (TC) water extract on liver in Rat. Treatment of TC water extract was orally administered 200, 300 mg/kg daily for one week and two weeks. The clinical parameters of serum, values of AST, ALT showed significantly higher than in normal group. Xanthine oxidase and aldehyde oxidase activities were significantly increased comparison with normal group. Microsomal enzymes, aminopyrine N-demethylase and aniline hydroxylase were not affected. Water extract of TC also increased hepatic rnalondialdehyde formation and reduced glutathione content. We also found that water extract of TC decreased activities of glutathione S-transferase and glutathione reductase but was not affected activities of $\gamma$-glutamylcysteine synthetase Thus, it seems that the water extract of TC induced decrease of oxygen free radical scavenger, glutathione content by inhibition of glutathione reductase which may reform oxidized glutathione to reduced glutathione.

Alcohol dehydrogenase 1 and NAD(H)-linked methylglyoxal oxidoreductase reciprocally regulate glutathione-dependent enzyme activities in Candida albicans

강사욱,곽민규 한국미생물학회 2021 The journal of microbiology Vol.59 No.1

Glutathione reductase (Glr1) activity controls cellular glutathione and reactive oxygen species (ROS). We previously demonstrated two predominant methylglyoxal scavengers– NAD(H)-linked methylglyoxal oxidoreductase (Mgd1) and alcohol dehydrogenase 1 (Adh1)–in glutathione-depleted γ- glutamyl cysteinyl synthetase-disrupted Candida albicans. However, experimental evidence for Candida pathophysiology lacking the enzyme activities of Mgd1 and Adh1 on glutathione- dependent redox regulation remains unclear. Herein, we have aimed to demonstrate that glutathione-dependent enzyme activities coupled with cellular ROS changes is regulated by methylglyoxal accumulation in Δmgd1/Δadh1 double disruptants. Δmgd1/Δadh1 showed severe growth defects and G1-phase cell cycle arrest. The observed complementary and reciprocal methylglyoxal-oxidizing and methylglyoxalreducing activities between Δmgd1 and Δadh1 were not always exhibited in Δmgd1/Δadh1. Although intracellular accumulation of methylglyoxal and pyruvate was shown in all disruptants, to a greater or lesser degree, methylglyoxal was particularly accumulated in the Δmgd1/Δadh1 double disruptant. While cellular ROS significantly increased in Δmgd1 and Δadh1 as compared to the wild-type, Δmgd1/Δadh1 underwent a decrease in ROS in contrast to Δadh1. Despite the experimental findings underlining the importance of the undergoing unbalanced redox state of Δmgd1/Δadh1, glutathione- independent antioxidative enzyme activities did not change during proliferation and filamentation. Contrary to the significantly lowered glutathione content and Glr1 enzyme activity, the activity staining-based glutathione peroxidase activities concomitantly increased in this mutant. Additionally, the enhanced GLR1 transcript supported our results in Δmgd1/Δadh1, indicating that deficiencies of both Adh1 and Mgd1 activities stimulate specific glutathione-dependent enzyme activities. This suggests that glutathione-dependent redox regulation is evidently linked to C. albicans pathogenicity under the control of methylglyoxal-scavenging activities.

글루타티온 강화 효모 배양물의 사료 내 첨가 급여가 육계 생산성, 혈액특성 및 계육 저장성에 미치는 영향

김동욱,노환국,이왕식,김상호 공주대학교 자원과학연구소 2019 자원과학연구 Vol.1 No.1

This experiment was conducted to investigate the effects of dietary supplementation of glutathioneenhanced yeast culture on growth performance, blood characteristics, and meat storage stability in broiler chicks. A total of six hundred 1-d-old male broiler chicks (Cobb) were divided into 5 groups with 4 replicates of 30 birds each. The five dietary treatments fed for 5 weeks were : NC (no antibiotics), PC (virginiamycin 10ppm and salinomycin 60ppm), and glutathione-enhanced yeast culture treated groups (0.1%, 0.3%, 0.5%). The final body weight and body weight gain were significantly increased in PC and glutathione-enhanced yeast culture treated groups compared to NC (p<0.05), and were linearly increased by increasing the level of supplemental glutathione-enhanced yeast culture (p<0.05). The feed intake in PC was significantly higher than other groups (p<0.05). The feed conversion ratio in all treated group was significantly improved as compared to that of NC (p<0.05). No significant difference among the all groups were observed on blood leukocyte profile. But total white blood cell (WBC), heterophil, lymphocyte, and stress indicator (heterophil : lymphocyte ratio) tended to be decreased by increasing the level of supplemental glutathione-enhanced yeast culture. The levels of blood urea nitrogen (BUN), creatinine, aspatate aminotransferase (AST), and alanine aminotransferase (ALT), which used as the blood biochemical parameters of liver and kidney damages were significantly decreased or tended to be decreased in glutathione-enhanced yeast culture treated groups than those of NC (p<0.05). The total antioxidant activity in blood serum was significantly higher in PC and glutathione treated groups than NC (p<0.05). And thiobarbituric acid reactive substances (TBARS) values of chicken breast meat during storage period were linearly decreased by increasing the level of supplemental glutathione-enhanced yeast culture (p<0.05). In conclusion, the dietary supplementation of glutathione-enhanced yeast culture improved the growth performance and health states by controling of reactive oxygen species toxicity. Furthermore, the glutathione-enhanced yeast culture decreased the chicken breast meat deterioration in relation to storage period. These results suggest the possibility that the glutathione-enhanced yeast culture could be used as a functional feed additive in broiler chicks.