Glutathione S-transferase activity in rats exposed to methyl parathion

Methyl parathion is an organophosphate insecticide that has been used in agriculture and the domestic sector for several years. This pesticide and others, arriving through different processes, exert significant effects on water quality with serious consequences for environmental and human health. The main objective of this study was to investigate the changes of Glutathione S-transferase enzyme activity in methyl parathion exposed rat tissues. For this purpose, wistar rats (Rattus norvegicus) were injected intraperitoneally with 7 mg kg^?1 dose of methyl parathion, while corn oil was applied to control groups in the same way. The liver, kidneys, brain and small intestine were quickly removed after 0, 2, 4, 8, 16, 32, 64, and 72 hours of injection of methyl parathion and the glutathione S-transferase activity was determined in these tissues. As a result it was seen that glutathione S-transferase activity increases in all tissues in the group of male and female rats to which methyl parathion was given. The increase of glutathione S-transferase activity may be a result of methyl parathion's toxic effect because it is one of the most important enzymes of detoxification metabolism.     

还原型谷胱甘肽调控shewanella oneidensis MR-1降解亚碲酸盐

探讨了生物活性小分子还原型谷胱甘肽（GSH）调控奥奈达湖希瓦氏菌（shewanella oneidensis）MR-1还原亚碲酸盐的特性及机理.结果表明,生物体系中加入0.1,0.4,1.0mmol/L GSH,与空白对照相比,亚碲酸盐的生物还原效率可分别提高55%、71%和78%,且GSH浓度在0.1~1.0mmol/L范围内与亚碲酸盐的生物还原效率呈正相关.采用单因素实验优化了培养条件,在温度35℃,pH 8.0,GSH浓度为0.4mmol/L的条件下,亚碲酸盐生物还原效率在24h内即可达到97%.采用6种不同呼吸抑制剂实验探讨亚碲酸盐生物还原的电子传递路径,确定GSH在亚碲酸盐生物还原电子传递链上的加速位点为NADH还原酶、甲萘醌和FAD脱氢酶.     

Toxicity of the mosquito control insecticide phenothrin to three life stages of the grass shrimp (Palaemonetes pugio)

Phenothrin is a synthetic pyrethroid used as a contact insecticide in mosquito control programs. This study compared the toxicity of phenothrin to adult, larval and embryonic grass shrimp (Palaemonetes pugio) and examined oxidative stress responses in adult and larval grass shrimp. The adult 24-h LC50 was 0.341 μg/L (95 % confidence intervals 0.282–0.412) and the 96-h LC50 was 0.161 μg/L (95 % CI 0.128–0.203 μg/L). The larval 24-h LC50 was 0.50 μg/L (95 % CI 0.441–0.568) and the 96-h LC50 was 0.154 μg/L (95 % CI 0.139–0.170 μg/L). In the presence of sediment, the 24-h LC50 was 6.30 μg/L (95 % CI 5.00–7.44 μg/L) for adults and 0.771 μg/L (95 % CI 0.630–0.944) for larvae. The sublethal biomarkers glutathione and lipid peroxidase (LPx) were examined after 96-h phenothrin exposure at five concentrations, and there were no statistically significant differences in these levels in adults or larvae compared to controls. There was a significant downward trend in larval LPx levels. This research confirms that phenothrin is highly toxic to grass shrimp and suggests that both adult and larval grass shrimp are appropriate life stages for risk assessments.     

氯代硝基苯胺对鲫鱼(Carassius auratus)血清抗氧化酶的影响

研究了2-氯-4-硝基苯胺、4-氯-3-硝基苯胺，2-氯-5-硝基苯胺对斑马鱼的急性毒性，96hLC50分别为6.99,2.58,8.63mg/L，为阐明氯化硝基苯胺类化合物对水生生物抗氧化酶的早期影响，将鲫鱼暴露于梯度浓度的2-氯-4-硝基苯胺，4-氯-3-硝基苯胺，2-氯-5-硝基苯胺中，研究鲫鱼血清SOD和GSH-PX活性短期（48h)内的变化，研究结果表明，在实验设置浓度下，随着暴露浓度升高，与空白对照组相比，3种化合物对SOD活性表现为先轻微激活后抑制，对GSH-PX活性先激活再抑制后有所回升，表明3种化合物对鲫鱼血清SOD和GSH-PX活性有显著影响，与3种化合物96hLC50相比，引起生化效应的暴露浓度明显降低，且反应快速，可考虑SOD和GSH-PX酶活性相结合作为该类化合物对水环境污染胁迫的敏感生物指示物。     

Cobalt and manganese stress in the microalga Pavlova viridis （Prymnesiophyceae）： Effects on lipid peroxidation and antioxidant enzymes

Pollution of marine environment has become an issue of major concern in recent years.Serious environmental pollution by heavy metals results from their increasing utilization in industrial processes and because most heavy metals are transported into the marine environment and accumulated without decomposition.The aim of the present study is to investigate the effects on growth,pigments, lipid peroxidation,and some antioxidant enzyme activities of marine microalga Pavlova viridis,in response to elevated concentrations of cobalt(Co)and manganese(Mn),especially with regard to the involvement of antioxidative defences against heavy metal-induced oxidative stress.In response to Co~(2 ),lipid peroxidation was enhanced compared to the control,as an indication of the oxidative damage caused by metal concentration assayed in the microalgal cells but not Mn~(2 ).Exposure of Pavlova viridis to the two metals caused changes in enzyme activities in a different manner,depending on the metal assayed:after Co~(2 )treatments,total superoxide dismutase(SOD)activity was irregular,although it was not significantly affected by Mn~(2 )exposure.Co~(2 )and Mn~(2 )stimulated the activities of catalase(CAT)and glutathione(GSH),whereas,ghitathione peroxidase(GPX)showed a remarkable increase in activity in response to Co~(2 )treatments and decreased gradually with Mn~(2 )concentration,up to 50μmol/L,and then rose very rapidly,reaching to about 38.98% at 200μmol/L Mn~(2 ).These results suggest that an activation of some antioxidant enzymes was enhanced,to counteract the oxidative stress induced by the two metals at higher concentration.     

荧光法测定黄瓜中的谷胱甘肽

研究了用荧光分光光度计测定黄瓜中谷胱甘肽的方法。在荧光条件激发波长EX为340nm,发射波长EM为420nm时,其测定回收率为99．55％－99．95％,变异系数为1．24％。     

流加发酵及添加L-半胱氨酸对产朊假丝酵母高密度培养合成谷胱甘肽的影响

通过7L罐考察了不同葡萄糖流加速率以及添加L-半胱氨酸对产朊假丝酵母(Candida utilis)WSH02-08高密度发酵生产谷胱甘肽(GSH)的影响.结果表明,在恒定速度(5.5g L-1h-1)流加葡萄糖30h的基础上,发酵45h后细胞干重最高达到73g L-1,此时向发酵罐中一次性添加L-半胱氨酸40mmol L-1,最终GSH产量和胞内GSH含量分别达到了1458mg L-1和2.26%.图4表1参11     

低浓度阿特拉津对鲫鱼谷胱甘肽-S转移酶(GSTs)活性的影响

研究不同作用时间和不同暴露浓度下阿特拉津对鲫鱼肝脏、肾脏和肌肉谷胱甘肽-S转移酶(GSTs)活性的影响.结果表明:阿特拉津对鲫鱼各个组织器官GSTs活性产生较强的影响.长期暴露下(24 d),阿特拉津对鲫鱼肝脏和肌肉GSTs活性基本表现为诱导作用,最大诱导率为110.81%和32.54%,且分别在0.1～5.0和1.0～10.0 mg·L-1质量浓度范围内存在剂量-效应关系;对鲫鱼肾脏GSTs活性具有抑制作用,最大抑制率为14.42%,且在所设质量浓度(0～10 mg·L-1)范围内均表现出剂量-效应关系.在10.0 mg·L-1质量浓度暴露下第6-14天内,阿特拉津与鲫鱼肝脏及肌肉GSTs活性间存在时间-效应关系;其他情况下,任何组织器官在试验期间均未表现出时间-效应关系.     

苯并［a］芘对黑鲷肝脏GST活性的影响及其与肝脏代谢酶和胆汁代谢产物之间的变化关系

为探讨苯并[a]芘(B[a]P)暴露对鱼类的影响并筛选特异、敏感的生物标志物,研究了B[a]P对黑鲷(Sparus macrocephalus)肝脏谷胱甘肽-S-转移酶(GST)活性的影响,并分析了肝脏7-乙氧基异吩唑酮-脱乙基酶(EROD)、GST活性与胆汁中B[a]P典型代谢产物3-羟基-苯并[a]芘(3-OHB[a]P)3者之间及与B[a]P之间的剂量、时间-效应关系.结果显示:在B[a]P(0.5、1.0、2.0和5.0μg·L-1)暴露期间,肝脏GST活性随暴露时间总体呈"钟"形的变化趋势,在第2d时达到峰值;在第12h、1d和2d时,GST活性与B[a]P暴露浓度呈显著正相关(R12h=0.966(p<0.05)、R1d=0.953(p<0.05)、R2d=0.824(p<0.05)).与前期研究结果对比分析发现,B[a]P(0.5、1.0、2.0和5.0μg·L-1)暴露7d时,胆汁中3-OHB[a]P浓度与B[a]P暴露浓度呈显著的剂量-效应关系(R=0.943,p<0.05);黑鲷肝脏GST、EROD活性与3-OHB[a]P的对数浓度均呈显著正相关(RGST=0.740(p<0.05)、REROD=0.839(p<0.05));2.0μg·L-1B[a]P暴露后,黑鲷肝脏EROD、GST活性与3-OHB[a]P随暴露时间的变化趋势基本相同,但并不完全一致.鱼类肝脏EROD、GST与胆汁中B[a]P代谢产物3者之间的变化关系较为复杂,暴露浓度和时间是影响其变化的重要因素,肝脏GST和EROD活性比胆汁中代谢产物更易受其影响,在B[a]P的环境监测中代谢产物可能是一种更为可行的生物标志物.     

Antioxidative responses of duckweed (<Emphasis Type="Italic">Lemna minor</Emphasis> L.) to short-term copper exposure

Goal, Scope and Background

Elevated concentrations of copper in the environment result in accumulation of the metal in plants and cause an increase in reactive oxidative species (ROS). The first response to elevated amounts of ROS is increased levels of enzymatic and non-enzymatic antioxidants that reduce oxidative stress. The aim of our study was to evaluate the early stages of antioxidative responses to the low copper concentrations usually present in moderately polluted environments. In addition, some other parameters were examined to evaluate the effect of copper on plants.

Methods

Duckweed (Lemna minor L.) was exposed to different concentrations of copper sulphate for up to 24 hours. Glutathione concentration and enzymatic activities of catalase, guaiacol peroxidase and glutathione reductase were measured spectrophotometrically. Additionally, delayed and prompt chlorophyll fluorescence was measured by luminometry and fluorometry, respectively. The accumulation of copper in plants exposed for 24 hours to various concentrations of copper sulphate was measured by flame atomic absorption spectrophotometry.

Results

The treatment of plants with copper sulphate resulted in an immediate decrease of the glutathione pool, which was replenished after 24 hours at CuSO₄ concentrations lower than 2 μM. Higher CuSO₄ concentrations caused a decrease of reduced glutathione. The responses of the antioxidant enzymes glutathione reductase, guaiacol peroxidase and catalase to CuSO₄ differed during the first six hours of exposure, but their enzyme activities all increased after 24 hours of exposure. All these enzymes displayed biphasic activity curves with maximum values between 0.5 μM and 1 μM CuSO₄. The response of guaiacol peroxidase was the most pronounced and statistically significantly specific and that of catalase the least. Delayed chlorophyll fluorescence decreased after exposure to 1 μM CuSO₄, but no significant effect on maximum quantum yield of photosystem II (Fv/Fm) was observed. L. minor accumulated relatively high concentrations of copper. The accumulation rate was higher at lower concentrations of copper in the test medium (up to 2 μM CuSO₄) than at concentrations above 2 μM CuSO₄.

Discussion

One of the most pronounced antioxidative responses to copper exposure was modified levels of oxidized and reduced forms of glutathione. The decrease of the glutathione pool is most probably coupled with induced production of phytochelatins. Antioxidative enzymes showed the biphasic enzyme activity characteristic of stress response. Guaiacol peroxidase exhibited the greatest significant increase of activity, even at higher CuSO₄ concentrations at which the activity of catalase and glutathione reductase dropped. The intensity of delayed chlorophyll fluorescence decreased, indicating reduced photosynthesis of plants under stress. All the measured parameters showed that plants respond to even low copper concentrations very soon after exposure. The accumulation rate of copper in duckweed tissues indicates that L. minor is an accumulator species.

Conclusions

The synchronized and prompt inducibility of antioxidants indicates their involvement in a general plant defence strategy for coping with metal-induced oxidative stress. Glutathione concentration and guaiacol peroxidase activity were found to be the most sensitive of the early indicators of exposure to copper concentrations present in polluted water bodies.

Recommendation and Perspectives

The experimental design of the present study allowed us to compare the sensitivity of various methods and parameters for detecting plant responses to heavy metal-induced oxidative stress. The level of glutathione and the enzyme activities of guaiacol peroxidase and glutathione reductase could be used as a rapidly determined early warning system in toxicity studies.