Regulation Of Glutathione S-transferase Research Articles

Changes in hepatic thiol contents and regulation of glutathione S-transferase by high-fructose diet: Effects of kefir and some probiotic bacteria

In this study, thiol/disulfide homeostasis in the liver tissues of high-fructose-fed rats was investigated in conjunction with the changes in the main hepatic detoxification enzyme, glutathione S-transferase (GST). Additionally, the effects of well-known probiotics namely Kefir, Lactobacillus helveticus, and Lactobacillus plantarum supplementation on the thiol/disulfate contents and GST activity and gene expression levels were analyzed. Fructose, administered as a 20% solution in drinking water for 15 weeks, developed an animal model of metabolic syndrome in male Wistar rats. Kefir, L. helveticus, and L. plantarum supplementations were given by gastric gavage once a day during the final 6-weeks. The changes in hepatic GST were determined with kinetic-optimized spectrophotometric enzyme assays and qRT-PCR. Total thiol, native thiol, and disulfide levels were analyzed using (5,5-dithio-bis-(2-nitrobenzoic acid) as a chromogenic agent. High-fructose consumption reduced total and native thiol contents while increasing disulfide levels in the liver tissues of rats. Kefir and L. plantarum normalized the thiol levels and all probiotics reduced disulfide contents. High fructose augmented total GST activity but reduced the GST-Mu isoform. L. helveticus and L. plantarum normalized the total and GST-Mu activity, respectively. These results demonstrated a shift toward disulfide formation in the hepatic tissues of rats fed with high fructose. A possible reason would be the increase in total GST activity that uses the free glutathione, the main native thiol source in cells, as a substrate. Besides, probiotics such as Kefir, L. helveticus, and L. plantarum have an improving effect on thiol/disulfide homeostasis as well as main detoxification enzymes.

Glutathione S-transferase gene diversity and their regulation by Nrf2 in Chinese mitten crab (Eriocheir sinensis) during nitrite stress

Eriocheir sinensis is one of the most important economic aquatic products in China. However, nitrite pollution has become a serious threat to the healthy culture of E. sinensis. Glutathione S-transferase (GST) is an important phase II detoxification enzyme, which plays a leading role in the cellular detoxification of exogenous substances. In this study, we obtained 15 GST genes (designated as EsGST1-15) from E. sinensis, and their expression and regulation in E. sinensis under nitrite stress were studied. EsGST1-15 belonged to different GST subclasses. EsGST1, EsGST2, EsGST3, EsGST4, and EsGST5 belonged to Delta-class GSTs; EsGST6 and EsGST7 are Theta-class GSTs; EsGST8 is a mGST-3-class GST; EsGST9 belonged to mGST-1-class GSTs; EsGST10 and EsGST11 belonged to Sigma-class GSTs; EsGST12, EsGST13, and EsGST14 are Mu-class GSTs; EsGST15 is a Kappa-class GST. Tissue distribution experiments showed that EsGSTs were widely distributed in all detected tissues. The expression level of EsGST1-15 was significantly increased in the hepatopancreas under nitrite stress, indicating that EsGSTs were involved in the detoxification of E. sinensis under nitrite stress. Nuclear factor-erythroid 2 related factor 2 (Nrf2) is a transcription factor that can activate the expression of detoxification enzyme. We detected the expression of EsGST1-15 after interfering with EsNrf2 in the hepatopancreas of E. sinensis with or without nitrite stress. Results showed that EsGST1-15 were all regulated by EsNrf2 with or without nitrite stress. Our study provides new information about the diversity, expression, and regulation of GSTs in E. sinensis under nitrite stress.

The regulation of glutathione s-transferases by gibberellic acid application in salt treated maize leaves.

The online version contains supplementary material available at 10.1007/s12298-022-01269-2.

Phytochromes and Their Role in Diurnal Variations of ROS Metabolism and Plant Proteome.

Plants are sessile organisms forced to adapt to environmental variations recurring in a day-night cycle. Extensive research has uncovered the transcriptional control of plants' inner clock and has revealed at least some part of the intricate and elaborate regulatory mechanisms that govern plant diel responses and provide adaptation to the ever-changing environment. Here, we analyzed the proteome of the Arabidopsis thaliana mutant genotypes collected in the middle of the day and the middle of the night, including four mutants in the phytochrome (phyA, phyB, phyC, and phyD) and the circadian clock protein LHY. Our approach provided a novel insight into the diel regulations, identifying 640 significant changes in the night-day protein abundance. The comparison with previous studies confirmed that a large portion of identified proteins was a known target of diurnal regulation. However, more than 300 were novel oscillations hidden under standard growth chamber conditions or not manifested in the wild type. Our results indicated a prominent role for ROS metabolism and phytohormone cytokinin in the observed regulations, and the consecutive analyses confirmed that. The cytokinin signaling significantly increased at night, and in the mutants, the hydrogen peroxide content was lower, and the night-day variation seemed to be lost in the phyD genotype. Furthermore, regulations in the lhy and phyB mutants were partially similar to those found in the catalase mutant cat2, indicating shared ROS-mediated signaling pathways. Our data also shed light on the role of the relatively poorly characterized Phytochrome D, pointing to its connection to glutathione metabolism and the regulation of glutathione S-transferases.

Microarray Data Analysis of as-induced Rice Roots by using Easygo and Mapman Softwares

Rice is one of the most important crops in Asian countries such as China, Vietnam... Many recent reports indicate that the arsenic content in rice exceeds the threshold and affects human health. Studying of molecular mechanisms and finding the arsenic resistance genes in rice which is extremely important and urgent. In this study, we analyzed the transcriptional changes of arsenic-treated rice root cells during 24 hours by microarray technique. Results showed that a large number of the differentially expressed genes (720 genes). EasyGO and Mapman softwares are powerful tools in analyzing microarray data and classifying functional groups as well as the important metabolic pathways in the cell. Results of microarray analysis using EasyGO showed that 74 down-regulated genes related to cellular component, 200 up-regulated genes involved in catalytic activity, 93 up-regulated genes involved in biological processes as responding to environmental stress, and 64 detoxification-realted genes are increased expression such as cytochrome P450, Glutathione-S-transferase and UDP-Glycosyltransferase. Mapman's microarray analysis reaults also indicate that numerous of arsenic-tolerance genes of rice roots. These results support for searching indicated genes in the selection of As-tolerance rice varieties.
 Keywords
 Asen, EasyGO, Mapman, microarray, Oryza sativa L.
 References
 [1] S.K. Panda, R.K. Upadhyay, S. Nath, Arsenic stress in plants. Journal of Agronomy and Crop Science 196 (2010) 161-174. https://doi.org/10. 1111/j.1439-037X.2009. 00407.x.[2] M.A. Rahman, H. Hasengawa, M.M. Rahman, M.A Miah, A. Tasmin. Arsenic accumulation in rice (Oryza sativa L.): Human exposure through food chain. Ecotoxicology and Environmental Safety 69 (2008): 317-324. https://doi.org/10. 1016/j.ecoenv.2007.01.005.[3] K.A. Marrs, The function and regulation of Glutathione S-transferase in plants. Plant Mol Biol 47 (1996) 127-58. https://doi.org/10.1146/ annurev.arplant.47.1.127.[4] L.M. DelRazo, B. Quintanilla-Vega, E. Brambila-Colombres, E.S. Caldero ́n-Aranda, M. Manno, A. Albores, Stress proteins induced by Arsenic. Toxicology and Applied Pharmacology 177 (2001)132-148. https://doi.org/10.1006/taap. 2001.9291.[5] T.L. Huang, Q.T.T. Nguyen, S.F. Fu, C.Y. Lin, Y.C. Chen, H.J. Huang, Transcriptomic changes and signalling pathways induced by arsenic stress in rice roots. Plant Molecular Biology 80 (2012) 587-608. https://link.springer.com/article/10.10 07/s11103-012-9969-z.[6] O. Thimm, O. Bläsing, Y. Gibon, A. Nagel, S. Meyer, P. Krüger, J. Selbig, L.A. Müller, S.Y Rhee, M. Stitt, Mapman: a user-driven tool to display genomics data sets onto diagrams of metabolic pathways and other biological processes. The Plant Journal 37 (2004) 914-939. https://doi.org/10.1111/j.1365-313X.2004. 02016.x.[7] J. Hartley-Whitker, G. Ainsworth, A.A. Meharg, Copper- and arsenate-induced oxidative stress in Holcus lanatus L. clones with differential sensitivity. Plant, Cell and Environment 24 (2001) 713-722. https://doi.org/10.1046/j.0016-8025.2001.00721.x.[8] S. Mishara, A.B. Jha, R.S. Dubey, Arsenite treatment induces oxidative stress, upregulates antioxidant system, and causes phytochelatin synthesis in rice seedlings. Protoplasma 248 (2011) 565-577. https://doi.org/10.1007/s00709-010-0210-0.[9] M. Chabannes, A. Barakate, C. Lapierre, J.M. Marita, Strong decrease in lignin content without significant alteration of plant development is induced by simultaneous down-regulation of cinnamoyl CoA reductase (CCR) and cinnamyl alcohol dehydrogenase (CAD) in tobacco plants, The Plant 28 (2001): 257-270. https://doi.org/10. 1046/j.1365-313X.2001.01140.x.[10] T. Goujon, V. Ferret, I. Mila, B. Pollet, Down-regulation of the AtCCR1 gene in Arabidopsis thaliana: effects on phenotype, lignins and cell wall degradability. Planta 217 (2003) 218-228. https://doi.org/10.1007/s00425-003-0987-6.[11] C. Li, S. Feng, Y. Shoa, L. Jiang, X. Lu, X. Hou, Effects of arsenic on seed germination and physiological activities of wheat seedlings. Journal of Environmental Sciences. 19 (2007) 725-732. https://doi.org/10.1016/S1001-0742(07) 60121-1.[12] A.A. Meharg, J. Harley-Whitaker, Arsenic uptake and metabolism in arsenic resistant and nonresistant plant species. New Phytologist 154 (2002) 29-43. https://doi.org/10.1046/j.1469-8137.2002.00363.x.
 
 

Glutathione S-Transferase Regulation in Calanus finmarchicus Feeding on the Toxic Dinoflagellate Alexandrium fundyense.

The effect of the dinoflagellate, Alexandrium fundyense, on relative expression of glutathione S-transferase (GST) transcripts was examined in the copepod Calanus finmarchicus. Adult females were fed for 5-days on one of three experimental diets: control (100% Rhodomonas spp.), low dose of A. fundyense (25% by volume, 75% Rhodomonas spp.), and high dose (100% A. fundyense). Relative expression of three GST genes was measured using RT-qPCR on days 0.5, 1, 2 and 5 in two independent experiments. Differential regulation was found for the Delta and the Sigma GSTs between 0.5 to 2 days, but not on day 5 in both experiments. The third GST, a microsomal, was not differentially expressed in either treatment or day. RT-qPCR results from the two experiments were similar, even though experimental females were collected from the Gulf of Maine on different dates and their reproductive output differed. In the second experiment, expression of 39 GSTs was determined on days 2 and 5 using RNA-Seq. Global gene expression analyses agreed with the RT-qPCR results. Furthermore, the RNA-Seq measurements indicated that only four GSTs were differentially expressed under the experimental conditions, and the response was small in amplitude. In summary, the A. fundyense diet led to a rapid and transient response in C. finmarchicus in three cytosolic GSTs, while a fourth GST (Omega I) was significantly up-regulated on day 5. Although there was some regulation of GSTs in response the toxic dinoflagellate, the tolerance to A. fundyense by C. finmarchicus is not dependent on the long-term up-regulation of specific GSTs.

Expression pattern of GSTP1 and GSTA1 in the pathogenesis of asthma

ABSTRACTReactive oxygen species (ROS) are known aggravating factors for airway inflammation in asthma. Glutathione S-transferases (GSTs) detoxify ROS and toxic compounds in environmental exposures. However, little is known about the regulation of GST and expression of GST subtypes in asthma. The aim of this study was to evaluate how GSTs are regulated in asthma. We observed total GST activity and expression of GST subtypes in murine asthma models and GST expressions in induced sputum cells of asthmatics. Total GST activity was increased in BAL fluids of OVA-treated murine asthma model. GSTP and GSTA are highly expressed in peribronchiolar mononuclear inflammatory cells and epithelial cells in OVA-treated mice. GSTM are expressed in epithelial cells in both OVA and PBS-treated groups. GSTP1 mRNA expression was increased in the lung of OVA-treated mice compared with PBS-treated mice. GSTA1, GSTM1, and GSTT1 mRNA expressions were not different between both groups. GSTA1 mRNA expression was increased in induced sputum cells of asthmatics compared with healthy controls. GSTP1, GSTM1, and GSTT1 mRNA expressions were not different between asthmatics and healthy controls. In asthmatics, GSTP1 and GSTA1 mRNA expressions were higher in induced sputum cells of asthmatics with PC20 ≤ 4 mg/ml than those with PC20 > 4 mg/ml. GSTM1 and GSTT1 mRNA expressions were not different between two groups. These findings suggest that GSTs are upregulated in the airways of asthmatics in response to increased oxidative stress. GSTP and GSTA are thought to play an important role in protecting the airways of asthmatics compared with GSTM and GSTT.

Interplay of β-Catenin with Xenobiotic-Sensing Receptors and its Role in Glutathione S-Transferase Expression

The Wnt/β-catenin pathway plays an important role in liver homeostasis, as well as during prenatal liver development, liver regeneration, and hepatocarcinogenesis. The connection of hepatic β-catenin activation and expression of drug-metabolizing enzymes has been established in the past few years: on the whole, a generally positive-regulatory effect of β-catenin on the expression and inducibility of many enzymes involved in phase I and phase II of drug metabolism has been described by different groups. The mechanisms underlying these processes are still not fully understood. However, there is accumulating evidence for a complex interaction of β-catenin with different xenobiotic-sensing receptors, which act as transcription factors after ligand activation, for example the aryl hydrocarbon receptor or the constitutive androstane receptor. Among others, these crosstalk mechanisms might explain the manifold effects of β-catenin on hepatic drug metabolism. In this review, the current knowledge regarding the role of Wnt/β-catenin signaling in the regulation of hepatic expression of glutathione S-transferases is presented. In addition, the crosstalk of β-catenin signaling with nuclear receptors involved in the regulation of glutathione S-transferases will be discussed.

Study of Early Leaf Senescence in Arabidopsis thaliana by Quantitative Proteomics Using Reciprocal 14N/15N Labeling and Difference Gel Electrophoresis

Leaf senescence represents the final stage of leaf development and is associated with fundamental changes on the level of the proteome. For the quantitative analysis of changes in protein abundance related to early leaf senescence, we designed an elaborate double and reverse labeling strategy simultaneously employing fluorescent two-dimensional DIGE as well as metabolic (15)N labeling followed by MS. Reciprocal (14)N/(15)N labeling of entire Arabidopsis thaliana plants showed that full incorporation of (15)N into the proteins of the plant did not cause any adverse effects on development and protein expression. A direct comparison of DIGE and (15)N labeling combined with MS showed that results obtained by both quantification methods correlated well for proteins showing low to moderate regulation factors. Nano HPLC/ESI-MS/MS analysis of 21 protein spots that consistently exhibited abundance differences in nine biological replicates based on both DIGE and MS resulted in the identification of 13 distinct proteins and protein subunits that showed significant regulation in Arabidopsis mutant plants displaying advanced leaf senescence. Ribulose 1,5-bisphosphate carboxylase/oxygenase large and three of its four small subunits were found to be down-regulated, which reflects the degradation of the photosynthetic machinery during leaf senescence. Among the proteins showing higher abundance in mutant plants were several members of the glutathione S-transferase family class phi and quinone reductase. Up-regulation of these proteins fits well into the context of leaf senescence since they are generally involved in the protection of plant cells against reactive oxygen species which are increasingly generated by lipid degradation during leaf senescence. With the exception of one glutathione S-transferase isoform, none of these proteins has been linked to leaf senescence before.

Isozyme- and gender-specific induction of glutathione S-transferases by flavonoids

Most dietary flavonoids have antioxidant activity in vitro however, secondary mechanisms such as the ability to influence gene expression with the consequent modulation of specific enzymatic activities involved in the intracellular response against oxidative stress, are being realized. In the following study, we examined the ability of the flavonoids: flavone, morin, naringenin, (+)-catechin, and quercetin to modulate the activity of glutathione S-transferases (GSTs) mGSTA, mGSTP and mGSTM in hepatic tissues of male and female Swiss Webster mice. Subchronic dietary exposure to morin, naringenin, (+)-catechin, and quercetin (2,500 mg/kg diet for 20 days) did not produce statistically significant changes in GST activity. Conversely, gender-, and isozyme-specific induction of mGSTs were observed in animals fed flavone. A sevenfold increase in total mGST activity was observed in female animals whereas a fourfold increase was observed in male animals. Enzyme specific assays indicate that there were greater increases of both mGSTM (eightfold) and mGSTP (fourfold) activities in females as compared to males (sixfold and twofold, respectively). As testosterone is involved in the regulation of GSTs in mice, castrated males were fed flavone for 5 days (2,500 mg/kg diet). In this case, dietary flavone resulted in similar fourfold increases in total GST activity in inact and castrated animals. Isozyme specific studies indicate that increases could be attributed to an induction of mGSTM and mGSTP.

Proteomic Analysis of Arabidopsis Glutathione S-transferases from Benoxacor- and Copper-treated Seedlings

Glutathione S-transferases (GSTs) are involved in many stress responses in plants, for example, participating in the detoxification of xenobiotics and limiting oxidative damage. Studies examining the regulation of this gene family in diverse plant species have focused primarily on RNA expression. A proteomics method was developed to identify GSTs expressed in Arabidopsis seedlings and to determine how the abundance of these proteins changed in response to copper, a promoter of oxidative stress, and benoxacor, a herbicide safener. Eight GSTs were identified in seedlings grown under control conditions, and only one, AtGSTU19, was induced by benoxacor. In contrast, four GSTs, AtGSTF2, AtGSTF6, AtGSTF7, and AtGSTU19, were significantly more abundant in copper-treated seedlings. The different responses to these treatments may reflect the potential for copper to affect many more aspects of plant growth and physiology compared with a herbicide safener. Differences between RNA and protein expression of GSTs indicate that both transcriptional and translational mechanisms are involved in regulation of GSTs under these conditions.

Pyruvate-induced long-term maintenance of glutathione s-transferase in rat hepatocyte cultures.

The addition of pyruvate to the culture medium has been reported to improve the maintenance of P450-dependent enzyme expression in primary rat hepatocyte cultures. In this study, the effects of 30mM pyruvate on cell morphology, albumin secretion and glutathione S-transferase (GST) expression were investigated as a function of the time in culture. The effect of triiodothyronine (T3) exposure on GST expression was also measured in pyruvate-treated cultures. Transmission electron microscopy showed that untreated hepatocytes deteriorated after culture for 7 days, whereas the morphology of the pyruvate-treated cells was similar to that observed in intact liver tissue. The albumin secretion rate was significantly higher in rat hepatocytes exposed to pyruvate than in control cells. In the presence of pyruvate, mu and alpha class GST activities were well maintained, whereas GST pi activity was increased over the entire culture period. HPLC analysis revealed that the complement of GST subunits present in hepatocytes is altered during culture with pyruvate: mu,class proteins remained relatively constant, whereas a decrease in the alpha class content was accompanied by a strong increase in GST subunit P1 (GSTP1). The induction of GSTP1 was confirmed at the mRNA level. In control cultures, pi class GST activity was increased, but total, mu, and alpha class GST activities continuously declined as a function of culture time and became undetectable beyond 7 days in culture. At the protein and mRNA levels, a much smaller increase in GSTP1 was observed than in the pyruvate cultures. When the pyruvate-treated cell cultures were exposed to T3, an inhibitory effect on GST activities and proteins was found. These results indicate that this simple culture model could be useful for studying the expression and regulation of GST.

Hydroxypropyl-β-cyclodextrin as delivery system for thyroid hormones, regulating glutathione S-transferase expression in rat hepatocyte co-cultures

Thyroid hormones play a role in the regulation of glutathione S-transferase (GST) expression. Here, co-cultures of rat hepatocytes with bile duct epithelial cells have been used to study the direct effects of both triiodothyronine (T3) and thyroxine (T4) on GST activities and proteins. Because T3 and T4 are poorly water soluble and organic solvents used to dissolve them often interfere with biotransformation pathways, an alternative delivery system namely hydroxypropyl-β-cyclodextrin (HPBC) has been applied. Appropriate control cultures contained either 0.02 or 0.10% (w/v) HPBC, the concentrations necessary to supply T3 and T4 (10 −9 to 10 −5 M) to the cells, respectively. No effect of the vehicle HPBC on the different GST isoenzyme activities and proteins could be observed. On the contrary, after 10 days of co-culture, T3 and T4 decreased GST protein concentrations as well as GST activities measured by 1-chloro-2,4-dinitrobenzene (broad spectrum), 1,2-dichloro-4-nitrobenzene (Mu class M1/M2-specific) and 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (Alpha class A1/2-specific) in a concentration-dependent manner. The Alpha class subunits A1/2 and A3, and the Mu class subunit M2 were mostly affected. No effect was observed on the Pi class enzyme. These findings indicate that a combination of co-cultured hepatocytes with an HPBC-based delivery system for hydrophobic compounds represents a powerful in vitro tool in drug development.

Selective and synergistic activity of L-S,R-buthionine sulfoximine on malignant melanoma is accompanied by decreased expression of glutathione-S-transferase.

L-buthionine-S,R-sulfoximine (BSO) selectivley inhibits glutathione (GSH) synthesis. Malignant melanoma may be uniquely dependent on GSH and its linked enzymes, glutathione S-transferase (GST) and GSH-peroxidase, for metabolism of reactive orthoquinones and peroxides produced during melanin synthesis. We compared the in vitro effects of BSO on melanoma cell lines and fresh melanoma specimens (n = 118) with breast and ovarian cell lines and solid tumors (n = 244). IC50 values (microM) for BSO on melanoma, breast and ovarian tumor specimens were 1.9, 8.6, and 29, respectively. The IC90 for melanoma was 25.5 microM, a level 20-fold lower than steady state levels achieved clinically. The sensitivity of individual specimens of melanoma correlated with their melanin content (r = 0.63). BSO synergistically enhanced BCNU activity against melanoma cell lines and human tumors. We followed GSH levels, GST enzyme activity, GST isoenzyme profiles and mRNA levels after BSO. BSO (50 microM) treatment for 48 hr resulted in a 95% decrease in ZAZ and M14 melanoma cell line GSH levels, and a 60% decrease in GST enzyme activity. GST-mu protein and mRNA levels were significantly reduced in both cell lines. GST-pi expression was unaffected. These data suggest that BSO action on melanoma may be related to GSH depletion, diminishing the capacity to scavenge toxic metabolites produced during melanin synthesis. We report here for the first time that BSO enhancement of alkylator action may be related in part to down regulation of GST. BSO may be a clinically useful adjunct in the treatment of malignant melanoma.

Regulation of glutathione S-transferases (GST) by interleukin 6 (IL6) and dexamethasone (DEX) . Department of Medicine, Emory University School of Medicine, Atlanta, Georgia

Regulation of glutathione S-transferases (GST) by interleukin 6 (IL6) and dexamethasone (DEX) . Department of Medicine, Emory University School of Medicine, Atlanta, Georgia

Regulation of glutathione S-transferase and quinone reductase gene expression

Regulation of glutathione S-transferase and quinone reductase gene expression

Regulation of glutathione S-transferases of Zea mays in plants and cell cultures

An antiserum to glutathione S-transferase (EC 2.5.1.18) from maize (Zea mays L.) responsible for herbicide detoxification has been raised in rabbit. The antiserum was specific to the Mr 26000 subunit of the enzyme from maize seedlings and suspension-cultured cells, and recognized the isoenzymes active toward both atrazine and metolachlor. When plants were treated for 24 h with the herbicide antidote N,N-diallyl-2-2-dich-loroacetamide (DDCA), enzyme activities toward metolachlor were doubled in the roots and this was associated with a 70% increase in immunodetectable protein. Translation of polysomal RNA in vitro showed that the increase in the transferase in root tissue was brought about by a ninefold increase in mRNA activity encoding the enzyme. Treatment of suspension-cultured cells with cinnamic acid, metolachlor and DDCA raised enzyme activities but did not increase synthesis of glutathione S-transferase. In cultured maize cells, enzyme synthesis was maximal in mid-logarithmic phase, coinciding with the highest levels of enzyme activity. When callus cultures were established from the shoots of a maize line known to conjugate chloro-s-triazines, enzyme activity towards atrazine was lost during primary dedifferentiation. However, levels of total immunodetectable enzyme and activity toward metolachlor were increased in cultured cells compared with the parent shoot tissue.