瑞香狼毒提取物对山楂叶螨的生物活性及酶活性影响

瑞香狼毒的氯仿、石油醚、甲醇提取物对果树重要害螨山楂叶螨有很好的内吸和触杀活性；对氯仿提取物处理后的成螨体内几种重要酶系活性测定发现，在内吸处理中，其对蛋白酶、谷胱甘肽－S－转移酶，酯酶同功酶有不同程度的激活作用；在触杀处理中，对蛋白酶、谷胱甘肽－S－转移酶影响不明显，对酯酶同功酶有一定的抑制作用；研究表明，瑞香狼毒可以有效杀死山楂叶螨，是一种很有利用价值的杀虫植物。     

Antioxidant processes are affected in juvenile rainbow trout (Oncorhynchus mykiss) exposed to ozone and oxygen-supersaturated water

Two groups of juvenile rainbow trout (Oncorhynchus mykiss) were exposed for 4 h to either hyperoxic (>40 mg O₂/l) or ozonised (5.2 μg O₃/l) water containing supersaturated oxygen. Gill, liver, blood cells and plasma were taken 1, 5, 12, 24 and 48 h following the exposure periods and antioxidant responses and oxidative damage studied in terms of, respectively, antioxidant enzyme activities/glutathione status and lipid peroxidation. Observed biochemical changes were significant at p<0.05. Ozonation elevated oxidised glutathione (GSSG) levels, increased lipid peroxidation and decreased reduced glutathione (GSH) levels in gills 1 h following exposure, which indicate oxidative stress. However, the longer-term effects (48 h) following ozone exposure resulted in increased GSH levels in both gills and liver and, measured as oxidative stress index (OSI), were indicative of enhanced potential of tissues to resist oxidative stress. Ozonation also resulted in elevated activity of superoxide dismutase (SOD) at 5–12 h following exposure in gills compared to 24–48 h in liver, indicating either differential regulation and time-courses of response, or an earlier impact of ozonolysis products on gills than liver. In contrast, catalase activity was elevated in both gills and liver at 24 h by the effects of hyperoxia alone. Hyperoxia also decreased GSSG levels in both gills and liver, but had no effect on lipid peroxidation. Increases were seen in the activities of selenium-dependent glutathione peroxidase as a result of ozonation (gills; 5 and 24 h) and hyperoxia (liver; 1 h), and in total GPX (Se-dependent and independent) as a result of ozonation (both tissues; 48 h), but no effects were seen on glutathione reductase (GR) in either tissues. Levels of GSH were increased in blood cells at various times following hyperoxia. Overall, the results show (i) differential antioxidant and pro-oxidant effects of ozonation compared to hyperoxia, (ii) gills and possibly blood as the first line of impact and defence, with later effects on liver, and (iii) that even after 24–48 h following ozone exposure, all antioxidant defences had not returned to pre-exposure values.     

柞蚕谷胱甘肽硫转移酶-theta基因(GSTT)的克隆及其在5龄幼虫丝腺中的表达规律

谷胱甘肽硫转移酶(GSTs)是一种解毒酶。为明确柞蚕谷胱甘肽硫转移酶-theta(GSTT)在柞蚕丝素蛋白准备和合成过程中所起的作用,采用生物信息学的方法克隆了柞蚕GSTT基因,并分析其在5龄幼虫丝腺中的表达规律。根据家蚕GSTT基因序列设计引物,获得了柞蚕GSTT基因的cDNA序列(651 bp,编码216个氨基酸,Gen-Bank登录号:EU541490)。该基因编码的蛋白与家蚕GSTT的同源性为89%。利用柞蚕18S核糖体蛋白基因作为内参,对柞蚕GSTT在不同时期的表达情况进行实时定量PCR检测,结果发现柞蚕GSTT的mRNA表达量在5龄第4天达到最高,后期逐渐下降。研究表明,在柞蚕丝素蛋白合成的5龄期,GSTT表达量大量提高,推测其主要功能为帮助柞蚕排除体内过多的氨基酸,以达到排毒目的。     

Quinone reductase (QR) inducers from Andrographis paniculata and identification of molecular target of andrographolide

In the present study, it was demonstrated that the petroleum extract of Andrographis paniculata (AP) had quinone reductase (QR) inducing activity, which might be attributed to the modification of key cysteine residues in Keap1 by Michael addition acceptors (MAAs) in it. To screen MAAs in AP, glutathione (GSH) was employed, and a LC/MS/MS method was implied. Three compounds, andrographoside, andrographolide, 14-deoxy-14,15-dehydroandrographolide were revealed could well conjugated with GSH. Then, andrographolide along with 4 new and 14 known compounds were isolated to conduct QR induction evaluation, and the CD (the concentration required to double the activity of QR) value of andrographolide is 1.43 μM. The QR induce activity of andrographolide might be attributed to its targeting multiple cysteine residues in Keap1, therefore, the alkylation of Keap1 by andrographolide was further studied and the result showed that four cysteine residues: Cys77, Cys151, Cys273 and Cys368 were alkylated, which indicated that Keap1 is a potential target for the QR induce activity of andrographolide.     

滞育发动前家蚕滞育性与非滞育性卵的谷胱甘肽氧化还原循环状态分析

为了调查滞育和非滞育性家蚕卵的谷胱甘肽氧化还原循环在滞育发动前是否存在差异,利用分光光度法检测滞育发动前(25℃中蚕卵产下后0～24 h)滞育与非滞育蚕卵的谷胱甘肽含量和相关代谢酶活性。滞育发动前,滞育性家蚕卵中的还原型谷胱甘肽(GSH)含量、氧化型谷胱甘肽(GSSG)含量、总谷胱甘肽(GSH+2GSSG)含量和GSH/GSSG比值变化不显著,谷胱甘肽S-转移酶(GST)活性变化不显著,但硫氧还蛋白过氧化物酶(TPX)活性下降23%,谷胱甘肽还原酶(GR)活性升高57%;非滞育性家蚕卵中的GSH含量及TPX和GST活性变化不显著,但GSSG含量升高61%,GSH+2GSSG含量升高41%,GSH/GSSG比值下降33%,GR活性下降16%。与非滞育性家蚕卵相比,滞育发动前滞育性家蚕卵中的GSH含量、GSSG含量和GSH+2GSSG含量较低,GSH/GSSG比值较高,TPX活性较低,GST活性无显著差异,GR活性较高。两种蚕卵中都未能检测到谷胱甘肽过氧化物酶(GPX)和硫氧还蛋白还原酶(TrxR)活性。结果表明,滞育发动前滞育性家蚕卵中较低的TPX活性和较高的GR活性共同导致较高的GSH/GSSG比值,使其谷胱甘肽氧化还原循环处于相对还原态。     

高效氯氟氰菊酯和阿维菌素对中华蜜蜂CarE和GSTs的影响

羧酸酯酶(Carboxylesterase,Car E)和谷胱甘肽S-转移酶(glutathione S-transferase,GSTs)是蜜蜂体内重要的生化标记酶。本研究探讨了高效氯氟氰菊酯lambda-cyhalothrin和阿维菌素abamectin亚致死剂量对中华蜜蜂Apis cerana cerana中肠Car E和谷GSTs活性的影响。结果表明,用阿维菌素和高效氯氟氰菊酯亚致死浓度LC5饲喂中华蜜蜂,明显诱导Car E活性增强;LC5浓度的阿维菌素诱导GSTs活性增强,而LC10~LC40的浓度抑制GSTs活性;高效氯氟氰菊酯LC5~LC40浓度对GSTs作用不明显;用LC10浓度的阿维菌素和高效氯氟氰菊酯处理中华蜜蜂72 h,Car E活性呈现先提高后降低的趋势,而对GSTs作用不明显。     

灌浆期高温对小麦旗叶中SOD和GR活性及相关基因表达量的影响

以山农23和济麦20为试验材料,研究灌浆期(花后10~20 d)高温对小麦旗叶中超氧化物歧化酶(SOD)和谷胱甘肽还原酶(GR)活性及相关基因表达量的影响。结果表明,在高温胁迫条件下,山农23的SOD活性一直显著高于对照,而济麦20的SOD活性变化呈先升高后降低的趋势。山农23中Fe-SOD和Mn-SOD表达量的变化与SOD活性的变化趋势相似,但Cu/Zn-SOD表达量的变化与SOD活性的变化趋势不同。济麦20中3个SOD基因表达量的变化均与SOD活性的变化基本一致。高温胁迫条件下两个小麦品种的GR活性均呈现先升高后降低的趋势,山农23中GR表达量的变化与GR活性的变化趋势基本一致,济麦20中GR表达量的变化早于GR活性的变化。总体来看,高温胁迫条件下山农23具有较强的抗氧化能力,Fe-SOD和Mn-SOD基因对SOD活性起主要作用,抗氧化酶相关基因对灌浆期高温胁迫的响应比酶活性更敏感。     

柽柳谷胱甘肽转移酶基因(ThGSTZ1)转入酵母的抗逆能力分析

[目的]研究盐、重金属、低温和高温胁迫下,柽柳GST基因（命名为ThGSTZ1）的抗性作用。[方法]将ThGSTZl基因构建到酵母表达载体pYES2中,转入酿酒酵母（Saccharomyces cerevisiae）获得重组型酵母,同时以转化空载体pYES2的重组酵母作为阴性对照;对2种重组酵母分别进行LiCl、NaCl、KCI、CdCl2、ZnCl2、MgCl2、-20和53qC胁迫处理,比较2种酵母的成活率。[结果]ThGSTZ1能有效的提高酵母的抗盐碱及极端温度的能力,表明ThGSTZ1可能参与柽柳的逆境胁迫应答过程。[结论]GST基因具有参与多种胁迫响应和调节的功能。     

Can enzymatic antioxidant defences in liver discriminate between wild and sea cage-reared Bluefin Tuna quality?

Bluefin Tuna (Thunnus thynnus L.) is a fish species of high nutritional and commercial value that is increasingly farmed in the Mediterranean basin. Oxidative stress research in aquaculture is particularly important in the assessment of both the health of farmed fish and seafood quality. Reactive oxygen species (ROS) are able to start the lipid peroxidation process which is potentially dangerous in fish, since they contain a high percentage of n-3 polyunsaturated fatty acids, in particular C20:5 n-3 and C22:6 n-3, which account for most of seafood's nutritional quality. The study of the antioxidant system and how the various components interact to delay post-harvest lipid peroxidation covers both the improvement of quality and the extension of shelf life of the fresh tuna product.In this study, hepatic antioxidant enzymes (catalase—CAT, superoxide dismutase—SOD and glutathione peroxidase—GPx) and lipid peroxidation levels (TBARS) of wild and sea cage-reared tunas were investigated. The aim was to examine the possible oxidative stress state of wild tunas killed by different fishing methods, compared with reared specimens exposed to different periods of confinement but killed by the same method.No significant differences between wild specimens were found in relation to catching techniques. A significant depletion was observed in liver SOD activity in reared tunas (5.00 U/mg protein) in comparison with wild specimens (5.90 U/mg protein). No differences in liver CAT activity between wild and reared tunas were found. In reared specimens we observed an oxidative stress state indicated by an increase in GPx activity (104.70 U/mg protein and 0.83 U/mg protein in reared and wild tunas, respectively). A significantly higher liver lipid content was observed in reared tunas, but no significant differences between wild and reared tunas in terms of TBA reactive substances content were found (2.03 vs 2.02 micromol TBARS/mg lipid). Our results suggest that the increased GPx activity leads to the suppression of lipid peroxidation in liver of reared tunas. This is the first time that antioxidant enzyme activities have been assayed in Bluefin Tuna and the stress state seems to be related to the diet of reared specimens rather than to their confinement, as indicated by the absence of differences in the antioxidant activities related to the period of confinement.     

杨树次生代谢物质对光肩星天牛羧酸酯酶和谷胱甘肽S—转移酶的影响

在昆虫与植物的相互作用关系中 ,次生代谢物质被看作是植物对昆虫的化学防御机制之一。然而 ,植食性昆虫能够在寄主植物上完成生长发育过程 ,其重要原因就在于虫体自身具有相应的对寄主植物次生代谢物质的代谢适应能力 (钦俊德 ,1 995 ;1 987)。昆虫对待寄主植物中有毒的次生代谢物质可通过羧酸酯酶 (Ｅ .Ｃ .3 1 1 2 )、谷胱甘肽转移酶 (Ｅ .Ｃ .2 5 1 1 8)、多功能氧化酶 (Ｅ .Ｃ .1 1 4 1 4 1 )等解毒酶 (Ｄａｕｔｅｒｍａｎｅｔａｌ.,1 988) ,将其转化为非毒性物质 ,从而使昆虫能够适应取食对象。在烟草夜蛾 (Ｈｅ ｌｉｏｔｈｉ…