3个湖泊中黄颡鱼（Pelteobagrus fulvidraco）体内CYP1A1和GST的比较及其功效分析

环境中污染物生物标志物的选取和研究是环境监测的重要内容之一,作为污染物肝脏解毒作用的关键酶,多功能氧化酶（CYP1A1）和谷胱甘肽硫转移酶（GST）常被用来作为多种污染物监测的生物标志物。比较了3个湖泊（巢湖、洪泽湖和鄱阳湖）黄颡鱼（Pelteobagrus fulvidraco）体内的CYP1A1和GST的活性,并对其进行功效分析。结果显示,3个湖泊中黄颡鱼体内CYP1A1和GST之间呈现反相关关系;洪泽湖黄颡鱼体内CYP1A1值明显小于其他两个湖,而洪泽湖黄颡鱼体内GST值明显大于巢湖,与鄱阳湖相比差     

Effect of E-cinnamaldehyde against Sclerotinia sclerotiorum on potato and induction of glutathione S-transferase genes

This research was carried out to assess the effect of E-cinnamaldehyde (EC) against Sclerotinia sclerotiorum, causal agent of potato white mold, under invivo and invitro conditions. Based on the results, EC was able to completely inhibit (P < 0.05) mycelial growth of four isolates in both volatile and contact phases after five days of inoculation at the concentrations 500 μl and 5 μl/ml, respectively. Moreover EC at concentrations 30 and 50 μl EC/ml sterile distilled water significantly reduced white mold on six potato cultivars including Pashandi, Istambouli, Agria, Marfauna, Alpha and Spartaan in greenhouse trials. In next step, induction of glutathione S-transferase (GST)-like genes identified from the pathogen (isolate 3) genome was assessed. After 2 h of exposure to both EC and cinnamom extract, four genes with locus numbers SS1G_07319.1, SS1G_07195.1, SS1G_00703.1 and SS1G_01918.1 were up-regulated. Furthermore, EC and cinnamon extract appeared to have no effect on gene expression in SS1G_10295.1. In this study, enzyme activity of GST was determined with 1-chloro-2, 4-dinitrobenzene as substrate. While enzymatic activity was 42.3 and 45.6 units mg⁻¹ protein in sterilized distilled water and ethyl acetate, respectively, it was recorded as 78.9 and 86.4 units mg⁻¹ protein for cinnamon extract and EC, respectively.     

靶标酶在除草剂研究与开发中的作用

大多数除草剂都是通过特殊酶的抑制而产生杀草作用的。根据作用靶标对除草剂进行分类,了解靶标酶的作用机理及特性,对于新型除草剂的设计和杂草抗性的防治能够起到很大的帮助。本文介绍了谷胱甘肽转移酶(GST)、细胞色素P450酶、乙酰乳酸合成酶(ALS)、乙酞辅酶A羧化酶(ACCace)的研究进展。并分别从酶的生理功能、酶学特征、抑制剂作用机理、抑制剂的研究、抗性等方面进行了不同程度的阐述。     

Effect of ginger supplementation on developmental toxicity induced by fenitrothion insecticide and/or lead in albino rats

Evaluation of the antioxidant and antiteratogenic role of ginger Zingiber officinale polyphenols against the toxicity induced by fenitrothion and/or lead in female albino rats were investigated. Adult virgin females were divided into 8 groups and were orally treated as follow: control (C), 1% w/w of ginger (G), 120 μg/animal lead as lead acetate (L), 10 mg/kg of fenitrothion (F), lead (120 μg/animal) fenitrothion (10 mg/kg) (LF), ginger (1%w/w) + fenitrothion (10 mg/kg) (GF), ginger (1%w/w) + lead (120 μg/animal) (GL), ginger (1%w/w) + lead (120 μg/animal) + fenitrothion (10 mg/kg) (GLF). Treatments were expanded for 28 days before pregnancy and during gestation period from zero to 6th day. Blood samples were taken at the day 20th of gestation and animals were sacrificed to investigate the effect of tested substances on dams and development of their fetuses. Inhibition in AchE in (F) and (LF) groups and elevation in plasma AchE in (L) groups were observed. Elevation in oxidative stress biomarker malondialdehyde (MDA) was recorded in all intoxicated groups concomitants with reduction in total reduced glutathione (GSH) and reduction in the activity of glutathione S-transferase (GST). Elevation in liver function biomarkers alanin amintransferase (ALT) and aspartate aminotransferase (AST) and reduction in plasma total protein and albumin were recorded in (F), (L) and (LF). Supplementation with ginger in diet attenuates the alteration in MDA, GSH, GST, ALT and AST, however, it failed to counteract the effect of F, L and LF on AchE, total protein and albumin. Significant alterations in maternal toxicity were recorded in (GF, GL, LF and GLF) compared with control group. Also, parameters of embryotoxicity and fetotoxicity indicated significant decrease in litter number that observed in F and L and the number of dead fetus/dam and litters number increased in L group. Supplementation with ginger decreased each of the number of died fetus, growth retardation and fetal length, while, it increased fetal weight. As regards to, teratological aspects, the percentage of skeletal malformations and visceral anomalies were observed in all feti obtained from treated groups with different percentages. Supplementation with ginger slightly attenuates the developmental toxicity of fenitrothion and/or lead.     

Effect of methyl parathion and chlorpyrifos on certain biomarkers in various tissues of guppy fish, Poecilia reticulata

Tests of acute toxicity were performed on the most common species of aquarium fish, Poecilia reticulata. Guppies (P. reticulata) were exposed to progressive concentrations of methyl parathion (MP) and chlorpyrifos (CPF); a semi-static method according to guidelines of OECD was used. Tests of acute toxicity were conducted using 10 fish for each separate concentration and for the control group. The results were subjected to probit analysis to determine the 96 h LC₅₀ values. The 96 h LC₅₀ values of MP and CPF to P. reticulata were 8.48 ppm/L (5.98–10.89) and 0.176 ppm/L (0.313–0.224) respectively. In addition, behavioral changes at each concentration were observed for the individual fish. Fish were exposed for 96 h to different sublethal concentrations of MP and CPF (¼ LC₅₀, 1/8 LC₅₀ and 1/10 LC₅₀) and their oxidative stress-induction potential was estimated in brain, liver and gills of fish. MDA content is induced in all tissues but maximum rise was observed in gills (161% and 153% for MP and CPF respectively). With regard to antioxidant defense system (ADS), GSH level decreased in the brain, liver and gills of tissues of MP treated fishes (22%, 6% and 13% respectively) and showed increase in brain and gills CPF treated (23% and 21% respectively). CAT, GST, GR and SOD levels fluctuated in all treatment groups relative to the control. Brain AChE showed dose-dependent inhibition in fish exposed to the higher concentrations reached 45% and 66% for MP and CPF respectively. Collective findings demonstrated that pesticide exposure of fish induced an increase in MDA and fluctuated ADS along with inhibited AChE. These findings may be used as valuable biomarkers for evaluation of water pollution.     

抗GST单克隆抗体的制备及应用

用纯化的重组GST(谷胱甘肽S-转移酶)蛋白免疫BALB/C小鼠,取其脾细胞与SP2/0骨髓瘤细胞融合,对杂交瘤细胞及时筛选,阳性孔经三次有限稀释法克隆,成功获得4B3和1H10共2株能稳定传代并分泌抗GST单克隆抗体(McAb)的杂交瘤细胞.2株单克隆抗体腹水间接ELISA效价达10-6,抗体类型均为IgG1,κ链,经Western-blot分析表明,2株单抗与GST-IBV-S1-F融合蛋白及重组GST蛋白均具有特异性反应,验证这2株单抗可用于检测GST融合蛋白的表达情况;用1H10单抗制备的亲和凝胶柱可用于GST融合蛋白的纯化.     

Cry8E亚致死浓度对马铃薯甲虫解毒酶和保护酶的影响

苏云金芽胞杆菌Cry8E杀虫晶体蛋白对马铃薯甲虫具有中等毒性,用亚致死剂量9和7μg/m L的Cry8E分别饲喂2、3和4龄的马铃薯甲虫幼虫,然后测定羧酸酯酶(Car E)、乙酰胆碱酯酶(ACh E)、谷胱甘肽-S-转移酶(GST)和过氧化氢酶(CAT)的酶活性。β-羧酸酯酶的活性随Cry8E浓度的升高明显提高,随龄期增长活性降低,酶活性在浓度间和龄期间差异都极显著。谷胱甘肽-S-转移酶活性下降,2龄幼虫体内的乙酰胆碱酯酶的活性明显升高,并且随龄期变化差异显著(P0.01)。3龄幼虫体内的CAT酶活性随Cry8E浓度的增加而显著升高(P0.01)。本研究阐明了Cry8E对马铃薯甲虫解毒酶和保护酶活性的影响,为理论研究奠定基础。