Shizukaol C alleviates trimethylamine oxide-induced inflammation through activating Keap1-Nrf2-GSTpi pathway in vascular smooth muscle cell.

BACKGROUND: Cardiovascular disease is one of the main causes of global mortality, and there is an urgent need for effective treatment strategies. Gut microbiota-dependent metabolite trimethylamine-N-oxide (TMAO) promotes the development of cardiovascular diseases, and shizukaol C, a natural sesquiterpene isolated from Chloranthus multistachys with various biological activities, might exhibit beneficial role in preventing TMAO-induced vascular inflammation. PURPOSE: The purpose of this study was to investigate the anti-inflammatory effects and the underlying mechanisms of shizukaol C on TMAO-induced vascular inflammation. METHODS: The effect and underlying mechanism of shizukaol C on TMAO-induced adhesion molecules expression, bone marrow-derived macrophages (BMDM) adhesion to VSMC were evaluated by western blot, cell adhesion assay, co-immunoprecipitation, immunofluorescence assay, and quantitative Real-Time PCR, respectively. To verify the role of shizukaol C in vivo, TMAO-induced vascular inflammation model were established using guidewire-induced injury on mice carotid artery. Changes in the intima area and the expression of GSTpi, VCAM-1, CD68 were examined using haematoxylin-eosin staining, and immunofluorescence assay. RESULTS: Our data demonstrated that shizukaol C significantly suppressed TMAO-induced adhesion molecule expression and the bone marrow-derived macrophages (BMDM) adhesion in vascular smooth muscle cells (VSMC). Mechanically, shizukaol C inhibited TMAO-induced c-Jun N-terminal kinase (JNK)-nuclear factor-kappa B (NF-κB)/p65 activation, and the JNK inhibition was dependent on the shizukaol C-mediated glutathione-S-transferase pi (GSTpi) expression. By further molecular docking and protein-binding analysis, we demonstrated that shizukaol C directly binds to Keap1 to induce Nrf2 nuclear translocation and upregulated GSTpi expression. Consistently, our in vivo experiment showed that shizukaol C elevated the expression level of GSTpi in carotid arteries and alleviates TMAO-induced vascular inflammation. CONCLUSION: Shizukaol C exerts anti-inflammatory effects in TMAO-treated VSMC by targeting Keap1 and activating Nrf2-GSTpi signaling and resultantly inhibits the downstream JNK-NF-κB/p65 activation and VSMC adhesion, and alleviates TMAO-induced vascular inflammation in vivo, suggesting that shizukaol C may be a potential drug for treating TMAO-induced vascular diseases.

Phytochelatin-Mediated Cultivar-Dependent Cd Accumulations of Lactuca sativa and Implication for Cd Pollution-Safe Cultivars Screening.

Cd pollution-safe cultivar (Cd-PSC) is a feasible strategy to minimize Cd contamination in leafy vegetables. The shoot Cd concentrations of 23 Lactuca sativa cultivars under Cd stress ranged from 0.124 to 2.155 mg·kg-1 with a maximum cultivar difference of 8 folds. Typical Cd-PSC C16 (L) and high-Cd-accumulating cultivar C13 (H) were screened to investigate the mechanisms of Cd accumulations in L. sativa through determining Cd concentrations, Cd subcellular distributions, phytochelatin profiles, and phytochelatin biosynthesis-related genes' expressions. Higher Cd distribution in a heat stable fraction in C13 (H) indicated that the high Cd accumulation trait of C13 (H) mainly depended on the Cd-phytochelatin complexes. Root phytochelatin concentrations were significantly elevated in C13 (H) (5.83 folds) than in C16 (L) (2.69 folds) (p < 0.05) under Cd stress. Significantly downregulated expressions of glutathione S-transferase rather than the regulation of phytochelatin synthesis genes in the root of C13 (H) might be responsible for sufficient glutathione supply for phytochelatins synthesis. These findings suggested that phytochelatin elevation in C13 (H) would favor the Cd root to shoot transportation, which provides new insights into the phytochelatin-related cultivar-dependent Cd accumulating characteristic in L. sativa.

Guhong injection mitigates myocardial ischemia/reperfusion injury by activating GST P to inhibit ASK1-JNK/p38 pathway.

BACKGROUND: Guhong injection (GHI), a novel compound preparation that is composed of a chemical drug, namely aceglutamide, and the aqueous extract of safflower (Carthamus tinctorius L.), exhibits extreme antioxidative, antiapoptotic, anti-inflammatory, and neuroprotective effects. Since oxidative stress, apoptosis, and inflammatory response are all the dominant mechanisms of myocardial ischemia/reperfusion (MI/R) injury, we probe into the protective mechanism of GHI on MI/R injury for the first time. METHODS: In this research, we first employed molecular docking to determine whether three active ingredients in GHI, acetylglutamine (NAG), hydroxysafflor yellow A (HSYA), and syringin, possessed the potential activity to modulate the protein, glutathione S-transferase P (GST P). We further identified the protective effect of GHI on myocardial tissue with TTC staining, HE staining, TUNEL staining, and ELISA, and on H9c2 with flow cytometry and ELISA. We next explored whether the cardioprotective effect of GHI on left anterior descending ligation-reperfusion in rats and hypoxia/reoxygenation (H/R) in H9c2 cells was related to activate GST P to inhibit ASK1-JNK/p38 pathway via approaches of qRT-PCR and Western blot. RESULTS: Results of molecular docking indicated that all three compounds spontaneously docked to GST P, among them the binding affinities of both HSYA and syringin to GST P were higher than NAG. In vivo, GHI reduced myocardial infarction size and mitigated myocardial pathological injury. In vitro, GHI enhanced cell viability and extenuated depolarization of mitochondrial membrane potential. In addition, the results of in vivo and in vitro studies demonstrated that the cardioprotection of GHI was associated with improving the mRNA and protein expression levels of GST P to modulate oxidative stress, and inhibiting the levels of mRNA expression and protein phosphorylation of ASK1, JNK, and p38. However, the suppressed effect of GHI on ASK1-JNK/p38 pathway was reversed by ethacrynic acid (EA, a GST inhibitor), indicating that the regulation of GHI on ASK1-JNK/p38 was related to the activity of GST P. Besides, the in vitro results of qRT-PCR and western-blot also certified that the inhibited JNK and p38 further reduced Bax expression and elevated Bcl-2 expression to reduce the expression of caspase-3 to exert anti-apoptosis effects. CONCLUSION: Taken together, the cardioprotection of GHI mainly incarnated in activating GST P to relieve oxidation properties, thereby inhibiting ASK1-JNK/p38 pathway to suppress apoptosis.

Deciphering the anti-filarial potential of bioactive compounds from Ocimum sanctum: a combined experimental and computational study.

CONTEXT: The anthelminthic effect of Ocimum species (Lamiaceae) has been reported, however, its anti-filarial effect has not been explored to date. OBJECTIVE: This study evaluates the effect of Ocimum sanctum L. (OS) against lymphatic filarial parasites. MATERIAL AND METHODS: The ethanol extract of OS (EOS) leaves was tested for anti-filarial activity against Setaria cervi. Equal size and number (n = 10) of adult female S. cervi worms were incubated in 125, 250 or 375 µg/mL EOS extract for 6 h at 37 °C. The OS bioactive components were identified by UPLC-ESI-MS/MS and subjected to docking and molecular dynamics (MD) simulation against filarial antioxidant proteins. RESULTS: The EOS significantly inhibited the motility of adult female S. cervi after 6 h of incubation. The motility was found to be reduced by 53.7% in 375 µg/mL and 43.8% in 250 µg/mL EOS after 6 h of treatment. The UPLC-ESI-MS/MS analysis of ethanol extract of O. sanctum revealed the presence of 13 bioactive compounds. The docking analysis showed eight OS bioactive compounds to have high binding affinity (> 4.8 kcal/mol) towards antioxidant proteins of filarial parasites. Additionally, MD simulation studies showed significant impact of (RMSD ≤ 10 Å) chlorogenic acid, luteolin and ursolic acid on filarial antioxidant enzymes/proteins. To our knowledge, this is the first report of the anti-filarial activity of Ocimum sanctum. DISCUSSION AND CONCLUSIONS: The effect of EOS and OS bioactive components on human filarial parasites can be further evaluated for the development of new anti-filarial formulations.

Ursolic acid alleviates tetrandrine-induced hepatotoxicity by competitively binding to the substrate-binding site of glutathione S-transferases.

BACKGROUND: Tetrandrine (TET), a bisbenzylisoquinoline alkaloid isolated from Stephania tetrandra S. Moore, is the only approved medicine in China for silicosis. However, TET-induced hepatotoxicity has raised safety concerns. The underlying toxic targets and mechanism induced by TET remain unclear; there are no targeted detoxification strategies developed for TET-induced hepatotoxicity. Ursolic acid (UA), a pentacyclic triterpene with liver protective effects, may have detoxification effects on TET-induced hepatotoxicity. PURPOSE: This study aims to explore toxic targets and mechanism of TET and present UA as a potential targeted therapy for alleviating TET-induced hepatotoxicity. METHODS: A TET-induced liver-injury model was established to evaluate TET toxicity and the potential UA detoxification effect. Alkenyl-modified TET and UA probes were designed to identify potential liver targets. Pharmacological and molecular biology methods were used to explore the underlying toxicity/detoxification mechanism. RESULTS: TET induced liver injury by covalently binding to the substrate-binding pocket (H-site) of glutathione S-transferases (GSTs) and inhibiting GST activity. The covalent binding led to toxic metabolite accumulation and caused redox imbalance and liver injury. UA protected the liver from TET-induced damage by competitively binding to the GST H-site. CONCLUSION: The mechanism of TET-induced hepatotoxicity is related to irreversible binding with the GST H-site and GST-activity inhibition. UA, a natural antidote, competed with TET on H-site binding and reversed the redox imbalance. This study revealed the hepatotoxic mechanism of TET and provided a targeted detoxifying agent, UA, to alleviate hepatotoxicity caused by GST inhibition.

Psoralen induces hepatotoxicity by covalently binding to glutathione-S-transferases and the hepatic cytochrome P450.

BACKGROUND: Psoraleae Fructus has been widely used in China and its surroundings; however, Psoraleae Fructus and its compound preparation have been reported recently to cause liver injury in clinics. Thus, its safe use has attracted increasing attention. The possible mechanism is related to the metabolism of psoralen, but it still needs further clarification. PURPOSE: The present study was designed to evaluate the toxicity of psoralen and investigate the potentially related molecular mechanisms using chemical biology methods combined with animal experiments to provide evidence for the rational clinical use of psoralen. METHODS: An in vivo experiment was conducted with a time series of 20-80 mg/kg psoralen to verify its toxic performance. Target capture and click reactions were used to investigate direct targets of psoralen. Selectivity for different glutathione-S-transferase (GST) subtypes in the liver and inhibition of cytochrome P450 (CYP450) were also detected. RESULTS: Psoralen build-up in the liver is the primary cause of liver damage. Our study revealed the mechanism by which psoralen induces liver injury. Psoralen can bind directly to CYP2D6, CYP3A4, GST-α, and GST-µ and inhibit their activities, causing the depletion of glutathione (GSH) in vivo, which in turn induces hepatic damage. The special structure of α,ß-unsaturated lactones in psoralen facilitates its attachment to its target; therefore, complementing psoralen with GSH can efficiently protect the liver from damage. CONCLUSIONS: Psoralen causes a disorder in drug metabolism by inhibiting the activity of CYPs and GSTs, causing exhaustion of GSH, and subsequently leading to liver damage. The co-administration of GSH and psoralen is an effective way to avoid liver injury in clinical settings.

Serum levels of vitamin A, selenium, and better dietary total antioxidant capacity are related to lower oxidative DNA damage: A cross-sectional study of individuals at cardiovascular risk.

The risk for cardiovascular diseases (CVR) has been associated with oxidative DNA damage, but the genetic and environmental factors involved in the antioxidant and DNA repair system contributing to this damage are unknown. The aim was to evaluate the levels of oxidative DNA damage in CVR subjects and how it is related with some genetic and nutritional factors. The cross-sectional study evaluated 136 individuals of both sexes, aged 20-59 years, with at least one cardiovascular risk factor. The global risk score was used to classify individuals at low, intermediate, and high cardiovascular risk. The dietary total antioxidant capacity (DTAC) was calculated using table with FRAP values. The oxidative DNA damage was verified by the comet assay. The variants null of Glutathione-S-transferases Mu1 and Theta 1(GSTM1 and GSTT1) and rs25487 of X-Ray Repair Cross Complementing Protein 1 (XRCC1) were analyzed by real-time PCR and PCR-RFLP, respectively. The oxidative DNA damage was higher in patients with intermediate/high CVR than in patients with low CVR (P=.01). Individuals with GSTT1/GSTM1 null genotypes or arg/gln+gln/gln genotypes of the XRCC1 (rs25487) gene showed similar levels of oxidative DNA damage compared wild genotype. Multivariate regression analysis demonstrated that oxidative DNA damage in individuals with CVR depends on serum levels of vitamin A, selenium, and DTAC independently of the other factors [F(6.110)=8.213; P<.001; R2=0.330]. These findings suggest that nutritional factors such as DTAC, vitamin A and selenium may have a protective effect against oxidative DNA damage in these individuals.

Glutathione Modulation in PVYNTN Susceptible and Resistant Potato Plant Interactions.

Glutathione is a metabolite that plays an important role in plant response to biotic stress through its ability to remove reactive oxygen species, thereby limiting the degree of potential oxidative damage. It can couple changes in the intracellular redox state to the development, especially the defense responses, of plants. Several studies have focused on measuring glutathione levels in virus infected plants, but have not provided complete information. Therefore, we analyzed, for the first time, the content of glutathione as well as its ultrastructural distribution related to susceptible and hypersensitive potato-Potato virus Y NTN (PVYNTN) interaction, with an aim of providing new insight into interactive responses to PVYNTN stress. Our findings reported that the inoculation of PVYNTN caused a dynamic increase in the content of glutathione, not only in resistance but also in susceptible reaction, especially at the first steps of plant-virus interaction. Moreover, the increase in hypersensitive response was much more dynamic, and accompanied by a significant reduction in the content of PVYNTN. By contrast, in susceptible potato Irys, the content of glutathione decreased between 7 and 21 days after virus inoculation, which led to a significant increase in PVYNTN concentration. Additionally, our findings clearly indicated the steady induction of two selected potato glutathione S-transferase StGSTF1 and StGSTF2 genes after PVYNTN inoculation, regardless of the interaction type. However, the relative expression level of StGSTF1 did not significantly differ between resistant and susceptible plants, whereas the relative expression levels of StGSTF2 differed between susceptible and resistant reactions. Therefore, we proposed that StGSTF2 can act as a marker of the type of response to PVYNTN. Our observations indicated that glutathione is an important component of signaling as well as the regulatory network in the PVYNTN-potato pathosystem. In resistance responses to PVYNTN, this metabolite activates plant defenses by reducing potential damage to the host plant cell, causing a reduction in virus concentration, while it can also be involved in the development of PVYNTN elicited symptoms, as well as limiting oxidative stress, leading to systemic infection in susceptible potato plants.

Genome-wide identification of glutathione S-transferase gene family members in tea plant (Camellia sinensis) and their response to environmental stress.

Glutathione S-transferases (GSTs) are ubiquitous enzymes involved in the regulation of plant growth, development, and stress responses. Unfortunately, the comprehensive identification of GSTs in tea plant has not been achieved. In this study, a total of 88 CsGSTs proteins were identified and divided into eight classes, among which the tau class was the largest. Chromosomal localization analysis revealed an uneven distribution of CsGSTs across the tea plant genome. Tandem duplication is the main force driving tea plant CsGSTs expansion. CsGSTs structures and conserved motifs were similar. The analysis of cis-regulatory elements in promoter regions showed that CsGSTs can response to multiple stresses, and that MYB may be involved in the transcriptional regulation of CsGST. RNA-Seq data revealed that the expression of most GSTUs was associated with various stresses, including pathogen and insect attack, cold spells, drought and salt stresses, nitrogen nutrition, bud dormancy, and morphological development, and the expression of these CsGSTs was obviously different in eight tissues. In addition, we proved that CsGSTU19, localized at the nucleus and cell membrane, was involved in tea plant defense against temperature stresses and Co. camelliae infection. These findings provide references for the further functional analysis of GSTs in the future.

Effects of selenium treatment on endogenous antioxidant capacity in signal crayfish (Pacifastacus leniusculus).

Selenium is an essential element that plays a role in numerous physiological processes and is critical for the maintenance of a strong endogenous antioxidant system. Previous work by our research group reported that the organophosphate pesticide dimethoate decreased glutathione S-transferase activity (GST) in signal crayfish (Pacifastacus leniusculus) collected from the Boise River (Idaho, USA). The goals of this study were to examine whether: 1) sodium selenite modulated the endogenous antioxidants glutathione (GSH), metallothionein (MT), and glutathione S-transferase (GST), thus suggesting a mechanism of antioxidant activity, 2) dimethoate exposure (pro-oxidant stressor) decreased GST activity in a localized population of signal crayfish collected from the Snake River (Idaho, USA), and 3) investigate whether selenium cotreatment ameliorated the adverse effects of dimethoate on GST activity due to the antioxidant properties associated with selenium. Selenium and dimethoate treatments (and co-treatments) did not modulate GSH or MT concentrations at the doses tested in this study. Furthermore, neither selenium nor dimethoate was factors influencing GST activity, and no interaction was found between the treatments. While our results did not support our predictions, they are suggestive and future studies examining the protective role of selenium in pro-oxidant exposure in this species are warranted. Population-specific responses as well as seasonal variations in endogenous antioxidant expression should be considered in future experiments.

Nanocidal Effect of Rice Husk-Based Silver Nanoparticles on Antioxidant Enzymes of Aphid.

Green synthesis of nanoparticles using plant-based extracts is momentously used in different fields of science because of their environment-friendly nature and cost-effectiveness. In the present study, silver nanoparticles were synthesized by using rice husk (non-toxic agricultural by-product) to determine their efficacy against aphid's (Sitobion avanae) mortality and antioxidant enzymes. UV-VIS spectroscopy of synthesized nanoparticles showed the maximum absorption peak at 440 nm, FTIR exhibited different peaks, and SEM confirmed the roughly spherical shape and 70-80 nm size of silver nanoparticles. Aphids were reared on wheat seedlings in the laboratory at 20-25 °C and 16:8 (light:dark) photoperiod. Insecticidal bioassays were conducted on aphids at three different concentrations (200 ppm, 400 ppm, 600 ppm) of nanoparticles for 2 days. Results showed the highest mortality of aphids being 93.3% at 600 ppm nanoparticle concentration after 2 days while the lowest mortality was observed at 200 ppm. Furthermore, the effect of silver nanoparticles on antioxidant enzymes was studied. Results of enzyme assays revealed that enzyme activities of catalase and glutathione-s-transferase increased in response to increased nanoparticle concentration. The current findings suggested that silver nanoparticles have probation for replacing commercially available insecticides for combating pests.

Oil-mediated oxidative-stress responses in a keystone zooplanktonic species, Calanus finmarchicus.

The copepod Calanus finmarchicus is an ecologically important species in the North Atlantic, Norwegian and Barents seas. Accidental or continuous petroleum pollution from oil and gas production in these seas may pose a significant threat to this low trophic level keystone species. Responses related to oxidative stress, protein damage and lipid peroxidation were investigated in C. finmarchicus exposed to a water-accommodated fraction (WAF) of a naphthenic North Atlantic crude oil. The exposure concentration corresponded to 50% of the 96 h LC50, and samples were obtained at 0, 24, 48, 72 and 96 h after exposure initiation. Gene expressions (superoxide dismutase, catalase, glutathione S-transferase, glutathione synthetase, heat shock protein 70 and 90, ubiquitin and cytochrome P-450 330A1), enzyme activities (superoxide dismutase, catalase, glutathione S-transferase) and concentrations of total glutathione and malondialdehyde were analyzed. Gene expression analyses showed no differences between controls and the exposed animals, however significantly higher glutathione S-transferase activity and malondialdehyde concentrations were found in the exposed group, suggests lipid peroxidation as main toxic effect.

Proanthocyanidins mitigate bile acid-induced changes in GSTT2 levels in a panel of racially diverse patient-derived primary esophageal cell cultures.

Persistent and symptomatic reflux of gastric and duodenal contents, known as gastroesophageal reflux disease (GERD), is the strongest risk factor for esophageal adenocarcinoma (EAC). Despite similar rates of GERD and other risk factors across racial groups, EAC progression disproportionately impacts Caucasians. We recently reported that elevated tissue levels of the detoxification enzyme GSTT2 in the esophagi of Blacks compared to Caucasians may contribute protection. Herein, we extend our research to investigate whether cranberry proanthocyanidins (C-PAC) mitigate bile acid-induced damage and GSTT2 levels utilizing a racially diverse panel of patient-derived primary esophageal cultures. We have shown that C-PACs mitigate reflux-induced DNA damage through GSTT2 upregulation in a rat esophageal reflux model, but whether effects are recapitulated in humans or differentially based on race remains unknown. We isolated normal primary esophageal cells from Black and Caucasian patients and assessed GSTT2 protein levels and cellular viability following exposure to a bile acid cocktail with and without C-PAC treatment. Constitutive GSTT2 levels were significantly elevated in Black (2.9-fold) compared to Caucasian patients, as were GSTT2 levels in Black patients with GERD. C-PAC treatment induced GSTT2 levels 1.6-fold in primary normal esophageal cells. GSTT2 induction by C-PAC was greatest in cells with constitutively low GSTT2 expression. Overall, C-PAC mitigated bile-induced reductions of GSTT2 and subsequent loss of cell viability regardless of basal GSTT2 expression or race. These data support that C-PAC may be a safe efficacious agent to promote epithelial fitness through GSTT2 induction and in turn protect against bile acid-induced esophageal injury.

Effect of Korean Red Ginseng on metabolic syndrome.

Metabolic syndrome (MS) refers to a clustering of at least three of the following medical conditions: high blood pressure, abdominal obesity, hyperglycemia, low high-density lipoprotein level, and high serum triglycerides. MS is related to a wide range of diseases which includes obesity, diabetes, insulin resistance, cardiovascular disease, dyslipidemia, or non-alcoholic fatty liver disease. There remains an ongoing need for improved treatment strategies for MS. The most important risk factors are dietary pattern, genetics, old age, lack of exercise, disrupted biology, medication usage, and excessive alcohol consumption, but pathophysiology of MS has not been completely identified. Korean Red Ginseng (KRG) refers to steamed/dried ginseng, traditionally associated with beneficial effects such as anti-inflammation, anti-fatigue, anti-obesity, anti-oxidant, and anti-cancer effects. KRG has been often used in traditional medicine to treat multiple metabolic conditions. This paper summarizes the effects of KRG in MS and related diseases such as obesity, cardiovascular disease, insulin resistance, diabetes, dyslipidemia, or non-alcoholic fatty liver disease based on experimental research and clinical studies.

The effect of NADPH oxidase inhibitor diphenyleneiodonium (DPI) and glutathione (GSH) on Isatis cappadocica, under Arsenic (As) toxicity.

The present work was conducted to assess the effects of arsenic (As, 1000 µM), diphenyleneiodonium (DPI, 10 µM) and reduced glutathione (GSH, 500 µM) on Isatis cappadocica. As treatment decreased plant growth and fresh and dry weight of shoot and root and also enhanced the accumulation of As. As stress also enhanced the oxidative stress biomarkers, hydrogen peroxide (H2O2) and malondialdehyde (MDA) content. However, the application of GSH decreased the content of H2O2 and MDA by 43% and 55%, respectively, as compared to As treatment. The antioxidants like superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX), glutathione reductase (GR) and glutathione S-transferase (GST) also enhanced with As stress. NADPH oxidase inhibitor, the DPI, enhances the effect of As toxicity by increasing the accumulation of As, H2O2, MDA. DPI also enhances the activity of antioxidant enzymes except GR and GST, However, the application GSH increased the plant growth and biomass yield, decreases accumulation of As, H2O2 and MDA content in As as well as As + DPI treated plants. The thiols content [total thiol (TT), non-protein thiol (NPT) protein thiols (PT), and glutathione (GSH)] were decreased in the As + DPI treatment but supplementation of GSH enhanced them. Novelty statement: The study reveals the beneficial role of GSH in mitigating the deleterious effects of Arsenic toxicity through its active involvement in the antioxidant metabolism, thiol synthesis and osmolyte accumulation. Apart from As, We provided the plants NADPH oxidase inhibitor, the diphenyleneiodonium (DPI), which boosts the As toxicity. At present, there is dearth of information pertaining to the effects of DPI on plants growth and their responses under heavy metal stress.GSH application reversed the effect of diphenyleneiodonium (DPI) under As stress preventing the oxidative damage to biomolecules through the modulation of different antioxidant enzymes. The application of GSH for As stressed soil could be a sustainable approach for crop production.

Nephroprotective effect of Artemisia herba alba aqueous extract in alloxan-induced diabetic rats.

BACKGROUND AND AIM: In the present study, we investigate the phytochemical composition and the nephroprotective effects as well as the antioxidant properties of Artemisia herba alba aqueous extract in alloxan-induced experimental diabetes in rats. EXPERIMENTAL PROCEDURE: Wistar rats were divided into four groups of seven rats each: Group I: Normal control (NC) received saline solution at 9‰ given by intraperitoneal way; Group II: Diabetic control (DC) received alloxan (150 mg/kg b.w) intraperitoneally; Group III: Normal control (NC + AHA) received saline solution at 9‰ and treated orally by AHA aqueous extract (400 mg/kg/b.w); Group IV: Diabetic control (DC + AHA) received alloxan solution (150 mg/kg b.w) intraperitoneally and treated by aqueous extract of AHA (400 mg/kg/b.w/day) orally after one week of alloxan administration. After 30 days, blood and tissue samples were collected for biochemical and histopathological analysis, respectively. Glomerular damage markers, including creatinine, serum urea, urine creatinine and urine urea levels were estimated. Creatinine clearance was also assessed. Oxidative stress parameters were assessed in the kidney homogenate. RESULTS AND CONCLUSION: Alloxan-exposure resulted in significant increase in blood glucose and serum level of glomerular damage markers. The antioxidant enzyme activities were significantly downregulated associated with an increase in malondialdehyde (MDA) level over the baseline values. Artemisia herba alba aqueous extract supplementation significantly improved the studied parameters. In concluding, the results obtained suggests that Artemisia herbs-alba aqueous extract supplementation reduces alloxan-induced free radical generation, potentiates the antioxidant defense system and alleviates renal sensitivity to oxidative stress.

Chromium Exposure in Late Gestation Period Caused Increased Levels of Cr in Brain Tissue: Association with Alteration of Activity and Gene Expression of Antioxidant Enzymes of F1 and F2 Generation Mice.

Chromium is a micronutrient which has found frequent use as supplements during pregnancy and could have a role in altering the antioxidant status in the brain. The present study was undertaken to estimate chromium levels in the brain, antioxidant enzyme activity with their gene expression, and learning and memory parameters on F1 and F2 generation mice when the F0 was exposed to chromium. The chromium levels in the brain were estimated using atomic absorption spectrophotometer. The enzyme activity of glutathione-s-transferase (GST) and catalase (CAT) was estimated and their gene expression was evaluated using RT-PCR. The spatial memory was tested using Morris water maze. The learning and recall memory was tested using the step down latency paradigm. The chromium levels were significantly raised in animals treated with Cr per se in F1 generation and quercetin cotreatment reduced the Cr levels in brain significantly. The enzyme activity of GST was significantly less in Cr-treated animals of both generations and this effect was significantly reversed on cotreatment with quercetin. The gene expression of GST matched the enzyme activity. However, catalase activity did not show significant decrease with Cr but cotreatment with quercetin resulted in significant decrease compared with control and this effect was not matched by its gene expression. We observed no significant change in learning and memory parameters in both generations following Cr exposure. Thus, this study demonstrates that chromium exposure in gestation causes changes in enzyme activity especially GST and this change was matched by change in gene expression in GST but not CAT. There was no effect on memory at the given dose.

Effect of lipoic acid supplementation on gene expression and activity of glutathione S-transferase enzyme in infertile men.

Oxidative stress has become the focus of interest as a potential cause of male infertility. We evaluate effects of lipoic acid (LA) supplementation on glutathione S-transferase (GST) expression. This randomized, triple-blind, placebo-controlled clinical trial was conducted on 44 infertile males with idiopathic asthenozoospermia. Men were randomized to receive 600 mg LA or placebo once daily for 12 weeks and semen samples and venous blood samples were obtained. GST expression, reactive oxygen species (ROS) levels, GST activity and reproductive hormone profiles were also measured. GST expression in the intervention group were significantly higher than the control group. Also, at the end of the study, GST activity increased, and ROS levels decreased significantly compared to the baseline. Additionally, the intervention group showed an increase in testosterone and decrease in serum follicle-stimulating hormone (FSH), luteinizing hormone (LH) and prolactin after 12 weeks, but this difference was not significant. We conclude a 12-week treatment with LA leads to improvements in reproductive hormones in serum, and significantly reduces the generation of ROS and increases the gene expression and activity of GST in seminal fluid.

Biofunctional Feed Supplemented With By-products of Olive Oil Production Improves Tissue Antioxidant Profile of Lambs.

BACKGROUND/AIM: Olive mill wastewater (OMW) is a byproduct of olive oil production. The aim of the study was to estimate the redox profile of lambs' vital organs after consumption of an OMW-supplemented feed. MATERIALS AND METHODS: Twenty-four lambs received breast milk until day 15. Then, they were divided in two groups: control and OMW, n=12 each. The control group received standard ration, while the OMW group received OMW enriched feed along with mother's milk until day 42 and animals (n=6 per group) were sacrificed. The remaining 12 received the feeds until day 70 and sacrificed. Tissue samples were collected at day 42 and 70 and specific redox biomarkers were assessed. RESULTS: Overall, the OMW feed improved tissue redox profile by affecting the glutathione S-transferase (GST) activity and γ-glutamate-cysteine ligase (γ-GCL) expression in all tested tissues. Superoxide dismutase (SOD) activity was not affected. CONCLUSION: The polyphenol-rich byproduct reinforced lamb redox profile and may putatively improve their wellness and productivity.

GSTM1 and Liver Iron Content in Children with Sickle Cell Anemia and Iron Overload.

Chronic blood transfusions in patients with sickle cell anemia (SCA) cause iron overload, which occurs with a degree of interpatient variability in serum ferritin and liver iron content (LIC). Reasons for this variability are unclear and may be influenced by genes that regulate iron metabolism. We evaluated the association of the copy number of the glutathione S-transferase M1 (GSTM1) gene and degree of iron overload among patients with SCA. We compared LIC in 38 children with SCA and ≥12 lifetime erythrocyte transfusions stratified by GSTM1 genotype. Baseline LIC was measured using magnetic resonance imaging (MRI), R2*MRI within 3 months prior to, and again after, starting iron unloading therapy. After controlling for weight-corrected transfusion burden (mL/kg) and splenectomy, mean pre-chelation LIC (mg/g dry liver dry weight) was similar in all groups: GSTM1 wild-type (WT) (11.45, SD±6.8), heterozygous (8.2, SD±4.52), and homozygous GSTM1 deletion (GSTM1-null; 7.8, SD±6.9, p = 0.09). However, after >12 months of chelation, GSTM1-null genotype subjects had the least decrease in LIC compared to non-null genotype subjects (mean LIC change for GSTM1-null = 0.1 (SD±3.3); versus -0.3 (SD±3.0) and -1.9 (SD±4.9) mg/g liver dry weight for heterozygous and WT, respectively, p = 0.047). GSTM1 homozygous deletion may prevent effective chelation in children with SCA and iron overload.