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Expression of Brassica napus L. γ-glutamylcysteine synthetase and Low-and High-Affinity Sulfate Transporters in Response to Excess Cadmium

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Journal of Integrative Plant Biology 2005年第2期47卷 243-250页

作者： XinSUN Xue-MeiSUN Zhi-MinYANG Shao-QiongLI JinWANG Song-HuaWANG DepartmentofBiochemistryandMolecularBiology CollegeofLifeSciencesNanjingAgriculturalUniversityNanjing210095China

In both the roots and leaves ofBrassica napus L. cv. Youyan No. 8 under treatment with 30 μmol/L Cd, massive production of non-protein thiols (NPT; mainly containing glutathione (GSH) and phytochelatins (PCs)) was induced, together with an increase in γ-glutamylcysteine synthetase (γ-ECS)mRNA transcripts. Because γ-ECS is the key enzyme catalyzing the first step in GSH biosynthesis, which, in turn, is converted to PCs, the Cd-induced increase in γ-ECS expression may be responsible for the observed increase in the production of NPT. Using a quantitative reverse transcription polymerase chain reaction (RT-PCR) approach, the expression of genes encoding a putative low-affinity sulfate transporter (LAST) and a putative high-affinity sulfate transporter (HAST) was determined at the transcriptional level. The RT-PCR analysis of relative transcript amounts indicates that the LAST gene in B. napus leaves showed a constitutive expression, which was hardly affected by Cd treatment. However, treatment with 30 μmol/L Cd for 2 or 3 d induced a marked increase in the expression of LAST in roots. Transcriptional expression of the HAST gene occurred in roots, but not in leaves. The expression of HAST only in the roots suggests that it has a specific function in sulfate uptake from soil and that the putative LAST may be responsible for the transport of sulfate from the roots to the shoots, as well as for the uptake of sulfate from soil. These results indicate that changes in transcriptional expression for sulfate transporters were required for the increased demand for sulfate during Cd stress.

关键词： Brassica napus L. cadmium γ-glutamylcysteine synthetase non-protein thiol sulfate transporters

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The Keap1/Nrf2 pathway regulatesγ-glutamylcysteine synthetase in chronic obstructive pulmonary disease in rats

The Keap1/Nrf2 pathway regulatesγ-glutamylcysteine syntheta...

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中国生理学会第23届全国会员代表大会暨生理学学术大会

作者： Jun-Li GAO,Ai-Guo DAI Department of Respiratory Medicine,Hunan Institute of Gerontology,Hunan Province Geriatric Hospital,Changsha 410001,China

＜正＞γ-glutamylcysteine synthetase（γ-GCS） plays an important role in the development of chronic obstructive pulmonary disease（COPD）.However,the mechanism that the Keap1/Nrf2 pathway regulates the gene expression ofγ-GCS is still not well *** may play a key role in the *** present study was undertaken to test the hypothesis that the Keap1/Nrf2 pathway and PERK induce the gene expression ofγ-GCS.A rat of COPD model was *** pulmonary function was aggravated in COPD *** COPD rats shared specific pathological features of *** situ hybridization demonstrated that the mRNA levels ofγ-GCS and Nrf2 were notably enhanced in COPD *** chemistry showed that the expressions of Nrf2,p-PERK andγ-GCS protein in COPD group were significantly higher than those in C *** expressions of Keap1 protein in two groups were ***/Cytosol fractionation and Western blot showed that Nrf2 protein was detected in both cytoplasm and nucleus and strongly increased in COPD ***1 protein was mainly detected in cytoplasm and was not significantly different in both groups.P-PERK protein was present mainly in cytoplasm and increased in COPD *** correlation analysis showed that Keap1 protein was negatively correlated with Nrf2 andγ-GCS,whereas p-PERK protein was positively correlated with Nrf2 andγ-GCS,respectively. Taken together,our results suggested that the Keap1-Nrf2 system and PERK can induce the gene expression ofγ-GCS in the pathogenesis of COPD.

关键词： γ-glutamylcysteine synthetase regulation pulmonary disease obstructive

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Different effects of nine clausenamide ennatiomers on liver glutathione biosynthesis and glutathione S-transferase activity in mice

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Acta Pharmacologica Sinica 2006年第8期27卷 1024-1028页

作者： Yu-qun WU Li-de LIU Hua-ling WEI Geng-tao LIU~1 Department of Pharmacology,Institute of Materia Medica,Peking Union Medical College & Chinese Academy of Medical Sciences.Beijing 100050.China Department of Pharmacology Institute of Materia Medica Peking Union Medical College & Chinese Academy of Medical Sciences Beijing China

Aim:To study the effects of nine synthetic clausenamide with different stereostructures on liver glutathione(GSH)biosynthesis and glutathione S-transferase(GST)activity in ***:The nine test compounds were racemic mixturesand their ennatiomers of clausenamide,neoclausenamide and *** were administered clausenamide 250 mg/kg once daily for 3 consecutivedays,ig,and were killed 24 h after the last *** mouse liver cytosol GSHand GST were determined with related biochemical ***:Nineclausenamides exhibited different effects on liver GSH and *** nineclausenamides,only(+)and(±)clausenamide markedly increased liver cytosolGSH *** mechanism of increasing liver GSH content of(+)clausenamideis mainly due to stimulating the key limiting enzyme γ-glutamylcysteine synthetase(γ-GCS)activity for GSH *** other test clausenamides had no sucheffect on liver *** of the nine clausenamides induced a significant increaseof GST ***:The effects of clausenamide ennatiomers on liverGST and GSH varied with the alterations of their spatial structures.(+)Clausena-mide stimulated liver GSH biosynthesis through enhancing γ-GCS activity.

关键词： clausenamide glutathione γ-glutamylcysteine synthetase glutathione S-transferase

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Activating PPARy Increases NQO1 and γ-GCS Expression via Nrf2 in Thrombin-activated Microglia

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Current Medical Science 2020年第1期40卷 55-62页

作者： Hang HANG Li-kun WANG Si-ying REN An-jun SONG Guo-feng WU Department of Emergency The Affiliated Hospital of Guizhou Medical UniversityGuiyang 550004China

The present study aimed to explore the molecular mechanisms underlying the increase of nicotinamide adenine dinucleotide phosphate:quinine oxidoreductase 1(NQO1)and y-glutamylcysteine synthetase(γ-GCS)in brain tissues after intracerebral hemorrhage(ICH).The microglial cells obtained from newborn rats were cultured and then randomly divided into the normal control group(NC group),model control group(MC group),rosiglitazone(RSG)intervention group(RSG group),retinoic-acid intervention group(RSG+RA group),and sulfbraphane group(RSG+SF group).The expression levels of NQO1,γ-GCS,and nuclear factor E2-related factor 2(Nrf2)were measured by real-time polymerase chain reaction(RT-PCR)and Western blotting,*** results showed that the levels of NQO1,γ-GCS and Nrf2 were significantly increased in the MC group and the RSG group as compared with those in the NC group(P<0.01).They were found to be markedly decreased in the RSG+RA group and increased in the RSG+SF group when compared with those in the MC group or the RSG group(P<0.01).The RSG+SF group displayed the highest levels of NQO1,γ-GCS,and Nrf2 among the five *** conclusion,a medium dose of RSG increased the anti-oxidative ability of thrombinactivated microglia by increasing the expression of NQO1 and γ-*** molecular mechanisms underlying the increase of NQO1 and γ-GCS in thrombin-activated microglia may be associated with the activation of Nrf2.

关键词： rosiglitazone peroxisome proliferator-activated receptor y nuclear factor E2-related factor 2 nicotinamide adenine dinucleotide phosphate:quinine oxidoreductase γ-glutamylcysteine synthetase

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Positive correlation between PPARγ,PGC-1αandγ-GCS in lungs of rats and patients with chronic obstructive pulmonary disease

Positive correlation between PPARγ,PGC-1αandγ-GCS in lung...

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中国生理学会第23届全国会员代表大会暨生理学学术大会

作者： Jie LI,Ai-Guo DAI,Rui-Cheng HU,Li-Ming ZHU,Shuang-Xiang TAN Department of Respiratory Medicine,Hunan Institute of Gerontology,Hunan Province Geriatric Hospital,Changsha 410016,China

＜正＞Oxidative stress is one of the major pathogenesis of chronic obstructive pulmonary disease（COPD）.γ-glutamylcysteine synthetase（γ-GCS） is one of the paramount antioxidant enzymes in *** proliferatoractivated receptor gamma（PPARγ） is a ligand-activated transcription factor,which is activated by specific ligands such as rosiglitazone（RGZ）,exerting multiple biological ***γcoactivator-1α（PGC-1α） is a PPARγcoactivator, which binds to PPARγby induction of PPARγligands,co-activating PPARγtarget *** evidence has suggested that PPARγ/PGC-1αcan regulate multiple antioxidant ***,the effect of PPARγ/ PGC-1αonγ-GCS during the development of COPD remains unclear *** measured the expression levels of PPARγ,PGC-1αandγ-GCS,γ-GCS activity and reactive oxygen species（ROS） contents in lungs of rats treated by cigarette smoke（CS）+lipopolysaccharide（LPS） and CS+LPS+RGZ,as well as lungs of patients suffering from *** with lungs from CS+LPS-treated rats,lungs of RGZ-treated rats demonstrated markedly lower ROS contents,and remarkable increase ofγ-GCS activity and increase of the expression levels of PPARγ,PGC-1αandγ-***,compared with controls,expression levels of PPARγ,PGC-1αandγ-GCS significantly increased in lungs of mild COPD patients,and progressively decreased in lungs of patients with moderate and severe COPD.γ-GCS protein was positively correlated with FEV1%.PPARγand PGC-1αproteins were positively correlated withγ-GCS activity and mRNA *** conclusion,γ-GCS showed compensatory up-regulation in the early stage of COPD,which progressively decompensate with increasing COPD *** activation of the PPARγ/PGC-1αpathway may protect against COPD progression by up-regulatingγ-GCS and relieving oxidative stress.

关键词： peroxisome proliferator-activated receptor gamma peroxisome proliferator-activated receptorγcoactivator-1 γ-glutamylcysteine synthetase chronic obstructive pulmonary disease

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甘蓝型油菜的修复能力-螯合系统对重金属耐受与累积反应和γ-谷氨酰转肽酶的合成（英文）

甘蓝型油菜的修复能力-螯合系统对重金属耐受与累积反应和γ-谷氨...

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第十二届国际油菜大会

作者： Katarzyna L.Janczur Przemysaw Nuc Konrad Wasinkiewicz Barbara Tomaszewska Department of Biochemistry Adam Mickiewicz University Poznań 61-614Poland Department of Biochemistry Adam Mickiewicz University Poznań 61-614Poland Department of Biochemistry Adam Mickiewicz University Poznań 61-614Poland Department of Biochemistry Adam Mickiewicz University Poznań 61-614Poland

世界范围的重金属污染对生态平衡和人类健康造成极大的威胁。应用植物的对重金属的高度耐受和积累的能力来净化重金属对土壤的污染,对生态系统进行修复越来越引起重视。植物的修复能力依赖于一种对胁迫激发的自卫机制。植物螯合系统中... 详细信息

世界范围的重金属污染对生态平衡和人类健康造成极大的威胁。应用植物的对重金属的高度耐受和积累的能力来净化重金属对土壤的污染,对生态系统进行修复越来越引起重视。植物的修复能力依赖于一种对胁迫激发的自卫机制。植物螯合系统中非蛋白硫醇（NPT）:谷胱甘肽（GSH）和螯合肽（PC）参与了重金属离子的吸收与螯合过程。在植物螯合系统对重金属胁迫反应中,螯合肽（一种短的富含半胱氨酸离子的结合肽）是非核糖体合成的,它来源于GSH,在生物合成途径中它以γ-谷氨酰转肽酶（ECS）作为限速酶。通过甘蓝型油菜品系DH O-120的重金属累积和螯合系统对铅胁迫的表现研究了它潜在修复能力。在试验中利用超过其他植物铅致死剂量（44375.2mg/kg,或48316.77mg/kg d.w.或3mM Pb（NO3）2）)证实了甘蓝型油菜具有良好的重金属耐受能力。99.8%的重金属位于根部,到达地上部分的铅很少。在3mM Pb（NO3）2的处理中,加入EDTA后可克服铅在植株中转移屏分枝中铅的含量增加了18%。对油菜幼苗和根部的PC和GSH生物合成动态进行了观察。24小时内铅的相对含量成比例增加至最大值。用RT-PCR和实时PCR检测了γ-ECS基因（GSH1）转录水平,发现该基因在根和茎中表达量同时伴随着酶活性的增加。甘蓝型油菜γ-ECS基因的序列已被测定,用拟南芥的GSH1结构预测该基因中的15个外显子,经分析它的启动子序列与***中的γ-ECS基因有74%同源。芸薹族的GSH1基因是多变的,通过RT-PCR研究了第一个外显子区域,结果显示该区域缺失了9个核苷酸。因此在Brassica napus基因组中可能至少有两个γ-ECS的等位基因。甘蓝型油菜具有相对高的生物学产量、较强的重金属耐受和累积能力,而其近缘种***已被证实的环境修复功能,使它作为环境修复植物的利用前景更为诱人。拟南芥的研究结果方便了油菜的分子分析,这使人们可以通过转基因来创造用于环境修复的油菜。最佳的品种可以通过修饰其螯合生物合成途径例如γ-ECS的过量表达等来获得。

关键词： Brassica napus phytoremediation lead phytochelatin system γ-glutamylcysteine synthetase

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