Modulating Antioxidant Enzyme Activities Research Articles

Unravelling the Influence of Chlorogenic Acid on the Antioxidant Phytochemistry of Avocado (Persea americana Mill.) Fruit Peel.

Oxidative stress is pivotal in the pathology of many diseases. This study investigated the antioxidant phytochemistry of avocado (Persea americana Mill.) peel. Different solvent extracts (dichloromethane, ethyl acetate, methanol, and water) of avocado peel were subjected to total phenol and flavonoid quantification, as well as in vitro radical scavenging and ferric reducing evaluation. The methanol extract was subjected to gradient column chromatographic fractionation. Fraction 8 (eluted with hexane:chloroform:methanol volume ratio of 3:6.5:0.5, respectively) was subjected to LC-MS analysis. It was assessed for cellular inhibition of lipid peroxidation and lipopolysaccharide (LPS)-induced ROS and NO production. The DPPH radical scavenging mechanism of chlorogenic acid was investigated using Density Functional Theory (DFT). The methanol extract and fraction 8 had the highest phenol content and radical scavenging activity. Chlorogenic acid (103.5 mg/mL) and 1-O-caffeoylquinic acid (102.3 mg/mL) were the most abundant phenolics in the fraction. Fraction 8 and chlorogenic acid dose-dependently inhibited in vitro (IC50 = 5.73 and 6.17 µg/mL) and cellular (IC50 = 15.9 and 9.34 µg/mL) FeSO4-induced lipid peroxidation, as well as LPS-induced ROS (IC50 = 39.6 and 28.2 µg/mL) and NO (IC50 = 63.5 and 107 µg/mL) production, while modulating antioxidant enzyme activity. The fraction and chlorogenic acid were not cytotoxic. DFT analysis suggest that an electron transfer, followed by proton transfer at carbons 3'OH and 4'OH positions may be the radical scavenging mechanism of chlorogenic acid. Considering this study is bioassay-guided, it is logical to conclude that chlorogenic acid strongly influences the antioxidant capacity of avocado fruit peel.

Silicon nanoparticles confer hypoxia tolerance in citrus rootstocks by modulating antioxidant activities and carbohydrate metabolism

Citrus is a remarkable fruit crop, extremely sensitive to flooding conditions, which frequently trigger hypoxia stress and cause severe damage to citrus plants. Silicon nanoparticles (SiNPs) are beneficial and have the potential to overcome this problem. Therefore, the present study aimed to investigate the effect of silicon nanoparticles to overcome hypoxia stress through modulating antioxidant enzyme activity and carbohydrate metabolism. Three citrus rootstocks (Carrizo citrange, Roubidoux, and Rich 16–6) were exposed to flooding (with and without oxygen) through different SiNP treatments via foliar and root zone. SiNPs applied treatment plants showed a significant increase in photosynthesis, leaf greenness, antioxidant enzymes, and carbohydrate metabolic activities, besides the higher accumulation of proline and glycine betaine. The rate of lipid peroxidation was drastically higher in flooded plants; however, SiNPs application reduced it significantly, ultimately reducing oxidative damage. Overall, Rich16-6 rootstock showed good performance via root zone application compared to other rootstocks, possibly due to genotypical variation in silicon uptake. Our outcomes demonstrate that SiNPs significantly affect plant growth during hypoxia stress conditions, and their use is an optimal strategy to overcome this issue. This study laid the foundation for future research to use at the commercial level to overcome hypoxia stress and a potential platform for future research.

Foliar application of glycine betaine to ameliorate lead toxicity in barley plants by modulating antioxidant enzyme activity and biochemical parameters

Lead (Pb) toxicity is a major problem in agricultural soil that negatively affects plant growth and development. Glycine betaine (GB) is an effective compatible solute that resists abiotic stress and plays an important role to mitigate various stresses. The present study is the first of its kind on the application of GB to mitigate Pb toxicity on barley cultivars. To elucidate the role of GB in mitigating Pb toxicity of three concentrations (15 mM, 25 mM, and 35 mM) in two barley varieties (BH-959 and BH-946) with and without foliar application of GB (2 mM) was examined. The study found that increasing Pb concentration significantly (p < 0.05) reduced the level of primary metabolites viz. photosynthetic pigments, protein, and carbohydrates in both cultivars upto 42.4%, 38.4% and 39% respectively. However malondialdehyde content, proline content, and antioxidant enzyme activity (SOD, CAT, and POX) were found to increased significantly (p < 0.05) as compared to control treatment upto 83.4%, 83.2% and 51% respectively. In contrast, the application of GB led to significantly (p < 0.05) improved physio-biochemical parameters as well as antioxidant enzyme activity (53%) and reduced oxidative stress along with malondialdehyde content (14.42%) in both varieties. An increment in these parameters revealed that exogenous application of GB (2 mM) significantly improves Pb (up to 35 mM) toxicity in barley plants and its use may be beneficial for crops susceptible to Pb toxicity to improve growth and yield.

Effects of Sihuang Zhili Granules on the Diarrhea Symptoms, Immunity, and Antioxidant Capacity of Poultry Challenged with Lipopolysaccharide (LPS).

A growing interest has been focused on Chinese herbs as alternatives to antimicrobial growth promoters, which are characterized by non-toxic side effects and drug resistance. The purpose of this study was to evaluate the effects of the Sihuang Zhili granule (abbreviated as Sihuang) on diarrhea, immunity, and antioxidation in poultry. Thirty male Leghorn chickens, aged 21 days, were randomly assigned to one of three groups with ten animals each. The control group (CON) received intraperitoneal saline injections, while the LPS-challenged group (LPS) and Sihuang intervention group (SH) received intraperitoneal injections of LPS (0.5 mg/kg of BW) and Sihuang (5 g/kg) at d 31, d 33, d 35, respectively. The control and LPS groups were fed a basal diet, while the SH group was fed a diet supplemented with Sihuang from d 21 to d 35. Analysis of the diarrhea index showed that the addition of Sihuang inhibited the increase in the diarrhea grade and the fecal water content caused by LPS, effectively alleviating poultry diarrhea symptoms. The results of the immune and antioxidant indexes showed that Sihuang significantly reduced the contents of the pro-inflammatory factors TNF- α and IL-1 β, as well as the oxidative stress markers ROS and MDA. Conversely, it increased the contents of the anti-inflammatory factors IL-4 and IL-10, along with the activities of antioxidant enzymes GSH-Px and CAT, thereby enhancing the immune and antioxidant abilities of chickens. Furthermore, Sihuang protected the chicken's ileum, liver, and immune organs from LPS invasion and maintained their normal development. In conclusion, this study confirmed the antidiarrheal effect of Sihuang in poultry farming and demonstrated its ability to improve poultry immunity and antioxidant capacity by modulating antioxidant enzyme activity and inflammatory cytokine levels.

Isolation and Characterization of Flavonoids from Fermented Dandelion (Taraxacum mongolicum Hand.-Mazz.), and Assessment of Its Antioxidant Actions In Vitro and In Vivo

Flavonoids are famous for their diverse sources, strong biological activity, and low toxicity and could be used as a natural antioxidant in animal husbandry. In this study, the purification process and antioxidant activity of flavonoids from fermented dandelion were investigated. The adsorption and desorption characterizations of AB-8 macroporous resin for flavonoids from fermented dandelion (FD) were determined and purification parameters were optimized. Qualitative analysis using UPLC-MS/MS analysis was explored to identify the components of the purified flavonoids of FD (PFDF). The antioxidant activity of PFDF in vitro and in vivo was analyzed. The optimum purification parameters were as follows: a sample concentration of 2 mg/mL, 120 mL of the sample volume, a pH of 2.0, and eluted with 90 mL of 70% ethanol (pH 5). After purification, the concentration of the flavonoids in PFDF was 356.08 mg/mL. By comparison with reference standards or the literature data, 135 kinds of flavonoids in PFDF were identified. Furthermore, PFDF had a strong reducing power and scavenging ability against 8-hydroxy radical and DPPH radical. PFDF can effectively reduce the oxidative stress of zebrafish embryos and IPCE-J2 cells by modulating antioxidant enzyme activities. In summary, the purified flavonoids from fermented dandelion have good antioxidant activity and display superior potential as a natural antioxidant in animal husbandry.

Ciprofloxacin and dexamethasone in combination attenuate S. aureus induced brain abscess via neuroendocrine-immune interaction of TLR-2 and glucocorticoid receptor leading to behavioral improvement

Staphylococcus aureus induced brain abscess is a critical health concern throughout the developing world. The conventional surgical intervention could not regulate the abscess-induced brain inflammation. Thus further study over the alternative therapeutic strategy for treating a brain abscess is of high priority. The resident glial cells recognize the invading S. aureus by their cell surface Toll-like receptor-2 (TLR-2). Glucocorticoid receptor (GR) was known for its immunosuppressive effects. In this study, an attempt had been taken to utilize the functional relationship or cross-talking between TLR-2 and GR during the pathogenesis of brain abscesses. Here, the combination of an antibiotic (i.e. ciprofloxacin) and dexamethasone was used to regulate the brain inflammation either in TLR-2 or GR blocking condition. We were also interested to figure out the possible impact of alternative therapy on behavioral impairments. The results indicated that combination treatment during TLR-2 blockade significantly reduced the bacterial burden and abscess area score in the infected brain. However, marked improvements were observed in anxiety, depression-like behavior, and motor co-ordination. The combination treatment after TLR-2 blocking effectively scavenged free radicals (H2O2, superoxide anion, and NO) through modulating antioxidant enzyme activities that ultimately control S. aureus induced glial reactivity possibly via up-regulating GR expression. The exogenous dexamethasone might regulate the GR expression in the brain by increasing the corticosterone concentration and the GC-GR mediated signaling. Therefore, this in-vivo study demonstrates the possible regulatory mechanism of bacterial brain abscess that involved TLR-2 and GR as a part of neuroendocrine-immune interaction.

Regulation of endogenous phytohormones alters the fluoranthene content in Arabidopsis thaliana

Phytohormones are crucial endogenous modulators that regulate and integrate plant growth and responses to various environmental pollutants, including the uptake of pollutants into the plant. However, possible links between endogenous phytohormone pathways and pollutant accumulation are unclear. Here we describe the fluoranthene uptake, plant growth, and superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and glutathione S-transferase (GST) activities in relation to different endogenous phytohormones and different levels in Arabidopsis thaliana. Three phytohormone inhibitors—N-1-naphthyl-phthalamic acid (NPA), daminozide (DZ), and silver nitrate (SN)—were used to regulate endogenous auxin, gibberellin, and ethylene levels, respectively. Fluoranthene inhibited plant growth and root proliferation while increasing GST and SOD activity. The three inhibitors reduced fluoranthene levels in Arabidopsis by either affecting plant growth or modulating antioxidant enzyme activity. NPA reduced plant growth and increased CAT activity. SN promoted plant growth and increased POD and CAT activity, whereas DZ increased POD activity.

Biomonitoring role of some cellular markers during heat stress-induced changes in highly representative fresh water mollusc, Bellamya bengalensis: Implication in climate change and biological adaptation

Owing to increasing concern of global climate-change, temperature rise is of great interest which can be primarily evaluated from the seasonal variations in some organisms. Aquatic environment can be extremely stressful to its inhabitants because most of them are poikilothermous. In the present study, attempt was made to evaluate the biological effects of oxidative-stress and adaptive/antioxidant capacities during temperature variations (36–40 °C for 24hrs to 72hrs) in Bellamya bengalensis both in environmental and laboratory conditions by testing some biomarkers like HSP70, catalase (CAT), superoxide dismutase (SOD), reduced glutathione (GSH) and glutathione reductase (GR). The biomarker potency of the molecules and the anti-oxidative metabolic-network was postulated and extrapolated to find its resemblance to the climate-change associated organismal variations. In a natural and eco-restored environment in the Eastern part of India, 10–20 fold increases in CAT, SOD and HSP70 protein expressions (Western blot results) were noticed in Bellamya paralleling to their increased enzymatic activities (gel zymogram studies) due to the seasonal (summer versus winter) temperature variation. It is evident from the consecutive three years’ study that this variation resulted in the unfavorable physico-chemical changes of water quality parameters like dissolved oxygen, biochemical oxygen demand, alkalinity and consequently decreased the animal density in summer. And that was revived due to their higher reproduction-rate in post rainy/winter season when temperature normalizes resulting in a restoration of favorable environment. In laboratory condition, the reduced GR and increased GPx indicated the oxidative damage as evident by higher tissue MDA level following to higher mortality. Changes in SOD and CAT activities suggest activation of physiological mechanism to scavenge the ROS produced during heat stress. However, when mortality increased at different time points (36 °C − 72 h and 38 °C − 72 h), these enzyme activities also decreased as they failed to save the tissues from ROS. The results suggest that temperature variation does alter the active oxygen metabolism by modulating antioxidant enzyme activities, which can be used as biomarker to detect sub-lethal effects of climate change-associated pollution. The parity in environmental and laboratory experimental results may justify this laboratory experiment as model heat-stress experiment and indicate temperature as a universal stressor which alone or in combination with other water parameters initiates a consistent adapting behavior. The Bellamya bengalensis being the highest faunal representative in its habitat may serve as a good bioindicator species.

Protective Effect of Silybum marianum and Silibinin on Endothelial Cells Submitted to High Glucose Concentration.

Silybum marianum Gaertn. (Milk thistle) has been used since ancient times for the relief of liver diseases characterized by intense oxidative stress such as inflammatory liver disease and cirrhosis. As oxidative stress by hyperglycemia is involved in micro- and macrovascular complications of type 2 diabetes, our aim was to assess the protective effect of milk thistle seed extract against oxidative stress induced by a high glucose concentration on endothelial cells (EA.hy926 cells). High-performance liquid chromatographic analysis shows flavonolignans silychristin and silibinin A and B as major components. No cell toxicity was observed for concentrations up to 100 µg/mL of milk thistle extract for 24 h. Concentrations of 5-25 µg/mL of the extract were used to assess the protective effect on EA.hy926 cells treated with 30 mM glucose for 24 h. Oxidative damage by 30 mM glucose was shown as a significant decrease in reduced glutathione and a significant increase in protein carbonyls and antioxidant enzyme activities. S. marianum extract recovered reduced glutathione and balanced the elevated carbonyls and enzyme activity. Silibinin alone also recovered reduced glutathione and antioxidant enzymes. S. marianum protects endothelial cell against oxidative damage by modulating antioxidant enzyme activity, reduced glutathione, and protein carbonyl levels.

Effect of polychlorinated biphenyls on oxidation stress in the liver of juvenile GIFT, Oreochromis niloticus.

The present study clearly showed that chronic exposure to polychlorinated biphenyls (PCBs) at environmentally relevant concentrations can damage juvenile tilapia livers by modulating antioxidant enzyme activities and gene transcription, which affects toxic bioaccumulation and histological congestion. The results suggest that PCBs caused a decrease in the activity of some hepatic antioxidative and biotransformation enzymes (SOD, CAT, GST, T-GSH, and MDA) in tilapia at 7 days, as well as transcriptional changes (sod, cat, and gst). Except for some antioxidant parameters (T-GSH, GSH/GSSG, T-AOC, and MDA), significant declines and increases occurred at 14 and 21 days, respectively. Most of the antioxidant enzymatic signatures and genotoxicity significantly increased at 14 and 21 days. This study presented evidence that PCBs could result in hepatic toxicity through oxidative stress in the early growth stages of tilapia, and we speculated that oxidative stress plays an important role in embryonic developmental toxicity induced by PCBs.

Silicon alleviates salt stress by modulating antioxidant enzyme activities in Dianthus caryophyllus ‘Tula’

Although silicon (Si) is not considered as an essential element, it is beneficial to the plant growth. Its effect is more evident under abiotic and biotic stress conditions. The objective of this study is to investigate the role of Si on the in vitro growth and resistance to salt stress of Dianthus caryophyllus ‘Tula’. The experiment was designed as a factorial design with 0, 50, or 100 mg·L−1 of potassium silicate (K2SiO3) in combination with 0, 50, or 100 mM sodium chloride (NaCl). The treatment of 50 mg·L−1 Si improved the growth of plant. However, the treatment of Si at 100 mg·L−1 reduced the growth. Although NaCl retarded the growth, addition of Si along with NaCl to the culture medium mitigated the effect of NaCl. A primary defense line by Si to overcome the photosynthetic depression was apparent from the increased chlorophyll content in the Si + NaCl treatment as compared to the treatment of NaCl alone. Enhancement of growth and resistance to salinity by Si was thought to be due to the modulation in activity of antioxidant enzymes, such as superoxide dismutase, ascorbate peroxidase, guaiacol peroxidase, and catalase. Therefore, our results suggested that 50 mg·L−1 Si supplementation could be optimal for improved growth in vitro and enhanced resistance against salinity in D. caryophyllus ‘Tula’.

Protective Effects of Black Rice Extracts on Oxidative Stress Induced by tert-Butyl Hydroperoxide in HepG2 Cells.

Black rice contains many biologically active compounds. The aim of this study was to investigate the protective effects of black rice extracts (whole grain extract, WGE and rice bran extract, RBE) on tert-butyl hydroperoxide (TBHP)-induced oxidative injury in HepG2 cells. Cellular reactive oxygen species (ROS), antioxidant enzyme activities, malondialdehyde (MDA) and glutathione (GSH) concentrations were evaluated as biomarkers of cellular oxidative status. Cells pretreated with 50 and 100 μg/mL of WGE or RBE were more resistant to oxidative stress in a dose-dependent manner. The highest WGE and BRE concentrations enhanced GSH concentrations and modulated antioxidant enzyme activities (glutathione reductase, glutathione-S-transferase, catalase, and superoxide dismutase) compared to TBHP-treated cells. Cells treated with RBE showed higher protective effect compared to cells treated with WGE against oxidative insult. Black rice extracts attenuated oxidative insult by inhibiting cellular ROS and MDA increase and by modulating antioxidant enzyme activities in HepG2 cells.

Protective effects of unsaponifiable matter from rice bran on oxidative damage by modulating antioxidant enzyme activities in HepG2 cells

Rice bran is a byproduct of the rice milling process. It contains high levels of bioactive phytochemicals such as phytosterols, tocopherols, tocotrienols, γ-oryzanol, triterpene alcohol, and other minor compounds, which have antioxidant and health-promoting properties. The objective of this study was to evaluate the protective effect of unsaponifiable matter (USM) from rice bran against oxidative damages induced by tert-butyl hydroperoxide (TBHP) in HepG2 cells. Levels of cellular reactive oxygen species (ROS), cellular lipid peroxidation, reduced glutathione (GSH), and antioxidant enzyme activity were evaluated as biomarkers of the cellular oxidative status. HepG2 cells were pretreated with various concentrations of USM (0–100 μg/mL) for 12 h prior to TBHP exposure. Pretreatment of HepG2 cells with USM prevented the cell damage induced by TBHP in a dose-dependent manner. Cellular generation of ROS, formation of malondialdehyde (MDA), and depletion of GSH induced by TBHP were prevented by pretreatment with USM. In addition, treatment with TBHP increased the activities of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase. However, pretreatment with USM significantly reduced the activities of these enzymes. These results indicate that treatment of HepG2 cells with USM confers significant protection against oxidative damage by modulating ROS production, GSH levels, and antioxidant enzyme activity.

Nitrate reductase-mediated nitric oxide production is involved in copper tolerance in shoots of hulless barley.

An NR-mediated early NO production in the shoots of hulless barley plays an important role in protecting hulless barley from Cu toxicity through enhanced antioxidant enzyme activities and antioxidant pools. Nitric oxide (NO) has been identified as an important signaling molecule that is involved in multiple plant physiological responses, especially under some abiotic stress. Here, we investigated NO production and its effects on copper (Cu) excess in hulless barley shoots. An early NO burst at 24h was observed in shoots of hulless barley, and the synthesis of early NO was mediated through nitrate reductase (NR), but not through nitric oxide synthase (NOS). Application of the NO donor sodium nitroprusside (SNP) efficiently alleviated Cu-induced shoot inhibition and decrease in chlorophyll content, as well as oxidative damage and reactive oxygen species (ROS) accumulation, while inhibiting NO accumulation by a specific NO scavenger or a NR inhibitor aggravated shoot inhibition as well as the increase of hydrogen peroxide (H2O2) content, supporting the role of an NR-mediated early NO production in hulless barley responses to Cu toxicity. Furthermore, elevated antioxidant enzyme activities were induced by Cu stress in the shoots of hulless barley and further significantly enhanced by NO donor, whereas suppressed by NO scavenger or NR inhibitor. On the other hand, the application of NO scavenger significantly reduced Cu-induced accumulation of glutathione (GSH) and ascorbate (Asc) in the shoots of hulless barley. Taken together, our results indicate that NO may induce hulless barley seedling tolerance to Cu toxicity through modulating antioxidant enzyme activity and antioxidants accumulation.

Exogenous sucrose increases chilling tolerance in cucumber seedlings by modulating antioxidant enzyme activity and regulating proline and soluble sugar contents

To determine the alleviative effects of exogenous sucrose on chilling stress, cucumber seedlings were pretreated with sucrose for 1 day and exposed to 15/8°C for 3 days. Among the sucrose concentrations, pretreatment with 50mM sucrose led to the highest levels of plant growth and caused the lowest levels of superoxide radical, hydrogen peroxide and malonaldehyde under chilling stress. The combined effects of 50mM sucrose pretreatment and chilling stress increased the activities of superoxide dismutase (EC 1.15.1.1), guaiacol peroxidase (EC 1.11.1.7), glutathione peroxidase (EC 1.11.1.9), ascorbate peroxidase (EC 1.11.1.11), glutathione reductase (EC 1.6.4.2), dehydroascorbate reductase (EC 1.8.5.1) and monodehydroascorbate reductase (EC 1.6.5.4) in leaves. This coincided with the increased transcript levels of copper/zinc superoxide dismutase, glutathione peroxidase and ascorbate peroxidase genes and correlated with the elevated contents of reduced glutathione and ascorbate. The combination of the sucrose pretreatment and chilling increased the contents of proline, soluble sugars, fructose, glucose and endogenous sucrose and enhanced the activities of soluble acid invertase (EC 3.2.1.26) and neutral invertase (EC 3.2.1.27) as well. We propose pretreatment with 50mM sucrose leads to higher contents of endogenous sucrose, thereby activating antioxidant enzymes and enhancing chilling tolerance in cucumber; the induction of proline, soluble sugars, soluble acid invertase and neutral invertase are also related to chilling stress mitigation by the sucrose pretreatment. Exogenous sucrose may have application possibility in nutrient solutions for a future trial of chilling reduction.

Growth, oxidative stress responses, and gene transcription of juvenile bighead carp (Hypophthalmichthys nobilis) under chronic-term exposure of ammonia.

Ammonia toxicity has become a universal problem for aquatic animals, especially fish. The purpose of the present study was to assess the chronic toxicity of ammonia to the juvenile bighead carp (Hypophthalmichthys nobilis). The authors measured the responses of growth performance (specific growth rate, condition factor, body weight, and body length), oxidative stress, and related gene transcription of juvenile bighead carp exposed to solutions with different concentrations of un-ionized ammonia (UIA; 0 mg L(-1) , 0.053 mg L(-1) , 0.106 mg L(-1) , 0.159 mg L(-1) , and 0.212 mg L(-1) ). The results showed that UIA had no effect on growth performance, glutathione content, or glutathione S-transferase gene transcription, but superoxide dismutase (SOD) activity was significantly elevated. In addition, different concentrations of UIA produced different degrees of damage to juvenile bighead carp: compared with control, lower UIA levels significantly decreased gene transcription of catalase (CAT) and increased malondialdehyde (MDA) levels; higher UIA concentration (0.212 mg L(-1) ) significantly increased gene transcription of the antioxidant enzymes CAT and SOD and reduced MDA levels. The data clearly demonstrate that chronic exposure of UIA at lower concentrations can result in some degree of impairment of antioxidative function, and chronic exposure at higher concentrations can enhance damage to juvenile bighead carp by modulating antioxidant enzyme activities and gene transcription.

Mercury Induced Biochemical Alterations As Oxidative Stress In Mugil Cephalus In Short Term Toxicity Test

Mugil cephalus juveniles of size 2.5 ±0.6cm were exposed to mercury in short term chronic toxicity test through static renewal bioassay to detect the possible biochemical agent as biomarkers in aquatic pollution and in estuarine contamination as specific. Lipid peroxidation levels, glutathione S-transferase, catalase, reduced glutathione and acetylcholinesterase were studied as biochemical parameters.Increased thio-barbituric acid reactive substances levels were observed under exposure to mercury, leading to the oxidative damage resulting in lipid peroxidation. Decreased activities of antioxidants, catalase and increased glutathione-S-transferase under short term chronic exposures to mercury were more prominent in metal stress suggesting activation of physiological mechanism to scavenge the ROS produced. Decreased values of reduced glutathione on prolonged exposures to mercury indicate utilization of this antioxidant, either to scavenge oxy-radical or act in combination with other enzymes. The acetylcholinesterase activity was found to be decreased during mercury exposure. The results also suggest that mercury alters the active oxygen metabolism by modulating antioxidant enzyme activities, which can be used as biomarker to detect sub-lethal effects in aquatic pollution.

Cinnamic acid pretreatment enhances heat tolerance of cucumber leaves through modulating antioxidant enzyme activity

To investigate the physiological mechanism of heat stress mitigated by exogenous cinnamic acid (CA), seedlings of a cucumber cultivar (Cucumis sativus cv. Jinchun no. 4) were pretreated with CA for 2d and then were exposed to normal (25/18°C) and elevated (42/38°C) temperatures for 3d. 50μM CA was chosen since it improved growth more than other CA concentrations under heat stress. At 2d, supply of 50μM CA elevated the activities of antioxidant enzymes superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6), guaiacol peroxidase (GPX, EC 1.11.1.7), glutathione peroxidase (GSH-Px, EC 1.11.1.9), ascorbate peroxidase (APX, EC 1.11.1.11), monodehydroascorbate reductase (MDHAR, EC 1.6.5.4), dehydroascorbate reductase (DHAR, EC 1.8.5.1) and glutathione reductase (GR, EC 1.6.4.2) in cucumber leaves, and it also increased the contents of reduced glutathione (GSH) and ascorbate (AsA). When the CA-pretreated cucumber seedlings have been exposed to heat stress, their antioxidant activities were changed further and were higher than heat treatment alone. Compared to heat treatment, the combination of CA pretreatment and heat also increased relative water content (RWC) of leaves, enhanced levels of proline and soluble sugars, elevated contents of CA and vanillic acid (VA) in leaves, and decreased levels of malonaldehyde (MDA), superoxide radical (O2−) and hydrogen peroxide (H2O2), while it resulted in 8.33% of leaf edges being dried up. Meantime, heat had adverse effects on plant growth, RWC, CA and VA contents in leaves, and it as well increased the levels of MDA, O2− and H2O2 and made the dried leaf edges to be 79.17%. We conclude that pretreatment with 50μM CA elevates CA and VA contents in leaves, enhances antioxidant activities under heat stress and decreases lipid peroxidation to some extent, accompanying with increasing proline and soluble sugar levels in heat-stressed leaves, thus the pretreatment enhances heat tolerance of cucumber seedlings.

Malaysian gelam honey reduces oxidative damage and modulates antioxidant enzyme activities in young and middle aged rats

Honey contains antioxidants such as phenolic compounds that prevent cellular oxidative damage that lead to aging, diseases and death. The aim of this study is to determine the effect of local gelam honey supplementation on oxidative status of young and middle-aged rats. Thirty-six male Sprague-Dawley rats were divided into two age groups; young (2 months) and middle aged (9 months). Each group was further divided into three groups; fed with plain water (control), supplemented with 2.5 and 5.0 g/kg of gelam honey for 30 days. DNA damage level was determined by comet assay, plasma malondialdehyde by high performance liquid chromatography and antioxidant enzymes activities (superoxide dismutase; SOD, glutathione peroxidase; GPx and catalase; CAT) were determined in the erythrocytes and liver. Results showed that supplementation of gelam honey reduced DNA damage, MDA level and GPx activity in the liver of both age groups. But erythrocytes GPx activity in young rats increased markedly with 5 g/kg of gelam honey supplementation. Liver and erythrocytes CAT activities decreased in both age groups when two different doses were used. Liver SOD activity also decreased in young rats supplemented with 5 g/kg of gelam honey. In conclusion, gelam honey reduces oxidative damage of young and middle aged rats by modulating antioxidant enzyme activities which was more prominent at higher concentration (5 g/kg body weight) compared to lower concentration (2.5 g/kg body weight). Key words: Gelam honey, antioxidant enzymes, DNA damage, malondialdehyde.

Effect of Tea Catechins on Regulation of Antioxidant Enzyme Expression in H2O2-Induced Skeletal Muscle Cells of Goat in Vitro

Skeletal muscle cells (SMCs) of goats were stress induced with 1 mM H(2)O(2) in the absence or presence of 0.5, 5, and 50 μg/mL tea catechins (TCs) incubation. Cells were harvested at 48 h postincubation with TCs to investigate the effects of TCs on cell proliferation, cell membrane integrity, antioxidant enzyme activities, and antioxidant enzyme genes and protein expression levels. Results showed that H(2)O(2) induction inhibited cell proliferation with or without TC incubation; moreover, the inhibition effect was enhanced in the presence of TCs (P < 0.001). H(2)O(2)-induced stress increased the lactate dehydrogenase (LDH) activity in the absence or presence of TC incubation, but concentrations of TCs, less than 5 μg/mL, showed protective functions against LDH leakage than in other H(2)O(2)-induced treatments. The catalase (CAT) activity increased when SMCs were stress induced with H(2)O(2) in the absence or presence of TC incubation (P < 0.001). H(2)O(2)-induced stress decreased CuZn superoxide dismutase (CuZn-SOD) and glutathione peroxidase (GPx) activities, whereas this effect was prevented by incubation with TCs in a concentration-dependent manner. H(2)O(2)-induced stress with or without TC incubation had significant effects on mRNA and protein expression levels of CAT, CuZn-SOD, and GPx (P < 0.001). CAT and CuZn-SOD mRNA expression levels were increased by different concentrations of TC incubation, and this tendency was basically consistent with corresponding protein expression levels. The GPx mRNA expression level increased with a low concentration of TCs but decreased with concentrations greater than 5 μg/mL of TCs, whereas GPx protein expression in all TC-incubated groups was lower than in the control treatment. The current findings imply that TCs had an inhibitory effect on cell proliferation and enhanced damage to the cell membrane integrity, but TCs affected antioxidant status in SMCs by modulating antioxidant enzyme activities at mRNA and protein expression levels.