氧化应激在硒缺乏致鸡淋巴细胞DNA损伤中的作用

为探讨氧化应激在硒缺乏致鸡淋巴细胞DNA损伤中的作用,选择1日龄健康海兰褐雏鸡120只,随机分为对照组和缺硒组,对照组饲喂硒含量为0.282 mg/kg的饲料,缺硒组饲喂硒含量为0.032 mg/kg的饲料,分别于30、45、60及75日龄采集胸腺、脾脏和法氏囊,检测硒含量、抗氧化功能和淋巴细胞DNA损伤.与对照组比较,缺硒组鸡胸腺、脾脏和法氏囊中硒含量降低;T-AOC,GSH-PX和SoD活性降低,MDA含量升高;淋巴细胞彗星率增加,并呈明显的时间-效应关系;胸腺、脾脏和法氏囊中硒含量、T-AOC、GSH-Px和SOD活性与淋巴细胞DNA损伤呈负相关,MDA含量与其呈正相关.结果表明,氧化应激与硒缺乏致鸡淋巴细胞DNA损伤密切相关.     

牦牛铜锌超氧化物歧化酶基因cDNA克隆测序及其分子进化研究

对牦牛Cu/Zn-SOD cDNA进行克隆测序。所扩增片段长度为842bp,编码区长度为459bp。扩增片段与奶牛Cu/Zn-SOD cDNA序列比较发现有16个碱基存在差异。通过与已知Cu/Zn-SOD cDNA序列的7个物种:人、猪、鼠、大鼠、奶牛以及马的cDNA序列的比较构建了分子进化系统树。对所得序列的ORF分析发现,牦牛Cu/Zn-SOD由152个氨基酸组成,并对氨基酸序列进行了比对分析。     

传染性法氏囊病沉淀抗体水平与免疫保护关系的研究

以传染性法氏囊病(IBD)基因工程重组亚单位油乳剂疫苗及其重组活载体疫苗实验室试制品免疫SPF鸡,以法氏囊眼观病变及法氏囊中IBDV抗原检测结果来评定免疫保护效力。结果血清IBDV沉淀抗体水平达到1/8的鸡能抵抗IBDV强毒的攻击,低于1/4的部分鸡不能抵抗IBDV强毒的攻击。经t检验,在以血清IBDV沉淀抗体水平≥1/8所得百分率、血清IBDV沉淀抗体水平≥1/4所得百分率、法氏囊眼观变化正常鸡只百分率及法氏囊中IBDV抗原检测的阴性率之间差异都不显著。表明以血清IBDV沉淀抗体水平≥1/4所得百分率与法氏囊中IBDV抗原检测的阴性率和法氏囊眼观变化正常鸡只百分率一样,适用于评定攻毒免疫保护效力。     

鸡传染性法氏囊病新型疫苗研究现状及发展趋势

鸡传染性法氏囊病(IBD)是由传染性法氏囊病病毒(IBDV)引起的鸡和火鸡的一种急性、高度接触性传染病。该病主要侵害3-12周龄的雏鸡与青年鸡,破坏法氏囊中的B淋巴细胞,导致不同程度的免疫抑制,从而使病鸡增加对并发和继发性病毒和     

鸡传染性法氏囊病的防治措施

鸡传染性法氏囊病是由传染性法氏囊病毒引起的一种高接触性传染病,主要破坏中枢免疫系统法氏囊中的B淋巴细胞,导致免疫抑制.该病发病突然,病程短,发病率高.     

LPS诱导不同品系雏鸡炎症反应相关基因的差异表达分析

分别以AA+肉仔鸡和海兰褐蛋雏鸡为研究对象,采用大肠杆菌脂多糖(LPS)连续3d腹腔注射11日龄雏鸡,收集血液、肝脏、胸腺、法氏囊、脾脏等组织,利用RT-PCR定量分析炎症反应相关基因akirin2、PPARγ和几种miRNA的转录活性变化。结果表明:在LPS接种肉鸡(AA+肉鸡)的肝脏、法氏囊和脾脏中akirin2基因转录活性均显著下调(P0.01),PPARγ基因在肝脏、法氏囊中转录水平也均显著下调(P0.01);在LPS接种蛋鸡(海兰褐鸡)的胸腺中akirin2基因转录活性显著上调(P0.01),但PPARγ基因在各组织中的转录水平差异均不显著;miR-146a(P0.05)、miR-21(P0.01)和miR-155(P0.01)在2个品系鸡的肝脏中表达均显著下调。研究不同品系鸡LPS诱导下相关基因转录活性的变化关系,对于深入了解不同品系鸡炎症反应分子机制的差异和开发炎症反应早期诊断的分子标记物具有重要的理论和实践意义。     

一种简易采摘鸡法氏囊的方法—背位法

鸡传染性法氏囊病病原为IDDV,在法氏囊中含量最高,在实际工作中经常涉及到采集法氏囊进行病毒分离等问题,通常采用的剖腹法比较繁琐,且易引起散毒和法氏囊被污染。法氏囊位于泄殖腔和尾荐椎之间,并与泄殖腔外壁腹膜通过结缔组织相连,其解剖     

我国鸡传染性法氏囊病活疫苗质量标准的某些评价

鸡传染性法氏囊病(Infectious bursal disease,IBD)又名腔上囊炎、传染性囊病,是由双RNA病毒科禽双RNA病毒属的传染性法氏囊病病毒(IBDV)引起的鸡和火鸡的一种急性、高度接触性传染病.该病主要侵害3～12周龄的雏鸡与青年鸡,破坏法氏囊中的B淋巴细胞,导致免疫抑制,使病鸡更易感其它致病因子,且对某些疫苗的免疫应答能力下降[1].     

传染性法氏囊病病毒变异的分子生物学研究进展

传染性法氏囊病（infectious bursal disease，IBD)是由传染性法氏囊病病毒(IBDV)引起的鸡和火鸡的一种急性高度接触性传染病。该病主要侵害3～6周龄雏鸡与青年鸡，以损害法氏囊中的淋巴细胞为特征；临床上出现增重减少、死亡、机体严重脱水和骨骼肌出血…。除直接引起鸡死亡与生产性能     

鸡Toll样受体在IBDV免疫中的作用研究

为了揭示鸡先天性免疫反应在IBDV疫苗过程中的作用,将40只14日海兰褐蛋鸡随机分成两组,实验组接种IBDV活疫苗,对照组注射生理盐水,隔离饲养。于免疫前和免疫后12h、24h、48h、72h、130h各取脾脏和法氏囊分离淋巴细胞并提取总RNA,用实时荧光定量检测方法检测各组中chTLR3、chTLR7、chTLR21表达的动态变化。结果显示,在免疫后36h,鸡法氏囊和脾脏中chTLR3表达变化最显著,在免疫后48h,鸡脾脏和法氏囊中chTLR21表达变化也较为明显。提示chTLR3在鸡对IBDV的免疫应答中起主要作用。     

野桑蚕铜锌超氧化物歧化酶基因的克隆分析与原核表达

超氧化物歧化酶是野桑蚕保护酶体系的重要酶类。利用RT-PCR方法克隆出野桑蚕铜锌超氧化物歧化酶(Cu/Zn-SOD)cDNA(EMB I登陆号:AM410997),其开放阅读框ORF长465 bp,编码154个氨基酸。同源性及系统进化分析表明,推导出的氨基酸序列与3种果蝇的同源性平均为69.3%,与线虫为57%,各物种中与Cu/Zn结合的残基高度保守。利用ExPASy的ScanProsite以及PSort和TMpred对此编码的蛋白质结构和功能域分析,预测出其具有2段Cu/Zn-SOD特异序列,且该蛋白不存在信号肽以及跨膜区。将Cu/Zn-SOD cDNA克隆到pET-28 a(+)表达载体,测序鉴定后以IPTG诱导表达,SDS-PAGE电泳鉴定其表达的融合蛋白质分子量为19.4 kD。     

Induction of the Expressions of Antioxidant Enzymes by Luteinizing Hormone in the Bovine Corpus Luteum

Luteoprotective mechanisms of luteinizing hormone (LH) involved in the maintenance of bovine corpus luteum (CL) function have not been completely clarified. Since antioxidant enzymes are well documented as antiapoptotic factors in the CL of many mammals, we hypothesized that the luteoprotective action of LH is mediated by stimulating the local production and action of antioxidant enzymes. To test the above hypothesis, in the present study, we examined the mechanisms involved in the luteoprotective actions of LH. Cultured bovine luteal cells obtained from the CL at the mid-luteal stage (days 8–12 of the estrous cycle) were treated with LH (10 ng/ml), onapristone (OP; a specific progesterone receptor antagonist, 100 μM) and diethyldithiocarbamate [DETC; an inhibitor of superoxide dismutase (SOD), 100 μM] for 24 h. LH in combination with or without OP significantly increased the mRNA and protein expressions of manganese SOD (Mn-SOD) and catalase (CATA) and SOD activity. While LH alone significantly increased the mRNA and protein expressions of SOD containing copper and zinc (Cu,Zn-SOD), OP in combination with or without LH significantly decreased the mRNA and protein expressions of Cu,Zn-SOD. In addition, Cu,Zn-SOD, Mn-SOD and CATA mRNA expressions were higher at the mid luteal phase than the other luteal phases. LH in combination with DETC significantly decreased LH-increased cell viability. The overall results suggest that LH increases cell viability by LH-increased antioxidant enzymes, resulting in maintenance of CL function during the luteal phase in cattle.     

干旱胁迫下AMF对枳生长和活性氧代谢的影响

在正常水分和干旱胁迫下,研究丛枝菌根真菌(AMF)Funneliformis mosseae对枳生长和活性氧代谢的影响。结果表明：6 周的干旱处理显著抑制了枳根系菌根侵染率和植株生长。接种AMF能显著增加正常水分和干旱胁迫下枳株高、茎粗、叶片数,也显著降低叶片丙二醛、过氧化氢和超氧阴离子自由基含量,并显著增强叶片超氧化物歧化酶(主要是Mn-SOD和Cu/Zn-SOD)、过氧化物酶和过氧化氢酶的活性。研究揭示了菌根化植株具有更高的抗氧化防御能力,导致较低的氧化伤害,从而增强植物的抗旱性。     

Antioxidant capacity and concentration of redox-active trace mineral in fully weaned intra-uterine growth retardation piglets

Background: The redox status of intra-uterine growth retardation(IUGR) piglets post-weaning has been poorly studied.Methods: Newborns from twenty-four sows were weighted, weaned at 21 d and fed a starter diet until sampling.Sampling was done at 14 d post-weaning. A piglet was defined as IUGR when its birth weight was 2 SD below the mean birth weight of the total population. At weaning, eighteen piglets with nearly equal body weight from each category(i.e. IUGR or normal birth weight(NBW) piglets) were selected and then allocated to two treatments,consisted of six replicates with each pen having three piglets.Results: Compared with NBW group, IUGR significantly decreased average daily gain(P 0.001), average daily feed intake(P = 0.003), and feed efficiency(P 0.001) of piglets during the first two weeks post-weaning. IUGR decreased the activities of total antioxidant capacity(P = 0.019), total superoxide dismutase(T-SOD, P = 0.023),and ceruloplasmin(P = 0.044) but increased the levels of malondialdehyde(P = 0.040) and protein carbonyl(P = 0.010) in plasma. Similarly, the decreased activities of T-SOD(P = 0.005), copper- and zinc-containing superoxide dismutase(Cu/Zn-SOD, P = 0.002), and catalase(P = 0.049) was observed in the liver of IUGR piglets than these of NBW piglets. IUGR decreased hepatic Cu/Zn-SOD activity(P = 0.023) per unit of Cu/Zn-SOD protein in piglets when compared with NBW piglets. In addition, IUGR piglets exhibited the decreases in accumulation of copper in both plasma(P = 0.001) and liver(P = 0.014), as well as the concentrations of iron(P = 0.002) and zinc(P = 0.048) in liver. Compared with NBW, IUGR down-regulated m RNA expression of Cu/Zn-SOD(P = 0.021) in the liver of piglets.Conclusions: The results indicated that IUGR impaired antioxidant capacity and resulted in oxidative damage in fully weaned piglets, which might be associated with the decreased levels of redox-active trace minerals. This study highlights the importance of redox status in IUGR offspring and provides a rationale for alleviating oxidative damage by dietary interventions aiming to supplement trace minerals and to restore redox balance in the future.     

小肽对大黄鱼仔鱼生长和小肠发育的影响及其在低温胁迫下的抗氧化应激效应

本试验分为生长试验和低温胁迫试验2部分。先分别投喂大黄鱼(Larimichthys crocea)仔鱼经不同浓度(0、0.5、1.0、2.0和3.0 mL/m 3)小肽营养强化后的轮虫和卤虫12 d,以探讨小肽对大黄鱼仔鱼生长和小肠发育的影响;再将大黄鱼仔鱼暴露在温度为12℃的水体中24 h,以探讨小肽对低温胁迫下大黄鱼仔鱼抗氧化能力和非特异性免疫力的影响。结果显示:不同浓度小肽均显著增加了轮虫和卤虫的必需氨基酸和总氨基酸含量(P<0.05),显著提高了大黄鱼仔鱼的体长及小肠绒毛高度、数量和组织面积(P<0.05)。小肽显著提高低温胁迫下大黄鱼仔鱼的谷胱甘肽(GSH)含量以及肝脏抗氧化酶[铜锌超氧化物歧化酶(Cu/Zn-SOD)、锰超氧化物歧化酶(Mn-SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶-1a(GPx-1 a)、谷胱甘肽过氧化物酶-1b(GPx-1 b)和谷胱甘肽还原酶(GR)]基因和非特异性免疫酶[c型溶菌酶(c-type LZM)、g型溶菌酶(g-type LZM)和碱性磷酸酶(AKP)]基因表达水平,显著降低活性氧(ROS)含量(P<0.05)。核转录因子NF-E2相关因子2(Nrf2)和核转录因子-κB(NF-κB)基因表达水平均与抗氧化酶基因和非特异性免疫酶基因表达水平呈显著正相关(P<0.05),并均与ROS含量呈显著负相关(P<0.05)。综上所述,小肽能够提高轮虫和卤虫的营养价值,从而改善大黄鱼仔鱼的生长和小肠发育;小肽通过诱导Nrf2和NF-κB的表达来增加抗氧化酶基因和非特异性免疫酶基因表达水平,从而缓解低温胁迫诱导的氧化损伤。     

柞蚕铜锌超氧化物歧化酶基因的克隆与表达分析

超氧化物歧化酶(SOD)是生物体内的重要保护性酶类。采用RT-PCR方法克隆了柞蚕(Antheraea pernyi)铜锌超氧化物歧化酶(Cu/Zn-SOD)基因的ORF序列。生物信息学分析表明,该序列共编码154个氨基酸,具有Cu/Zn-SOD的保守性结构特征,与家蚕(Bombyxmori)、美国白蛾(Hyphantria cunea)、野桑蚕(Bombyx mandarina)以及果蝇(Drosophila melanogaster)Cu/Zn-SOD基因的同源性分别是81.5%、81.7%、81.5%、66.7%。柞蚕蛹经紫外线照射处理后用半定量PCR方法检测,发现照射不同时间脂肪体内Cu/Zn-SOD基因的转录水平存在差异,在照射5 min后开始增加,至10 min时为转录高峰,15 min时又呈下降趋势。     

Assessment of degree of oxidative stress and antioxidant concentrations in dogs with idiopathic dilated cardiomyopathy.

OBJECTIVE: To assess degree of oxidative stress and antioxidant concentrations in dogs with idiopathic dilated cardiomyopathy (IDCM). DESIGN: Prospective study. ANIMALS: 18 dogs with IDCM and 16 healthy control dogs. PROCEDURE: Concentrations of malondialdehyde (an indicator of oxidative stress); vitamins A, C, and E; glutathione peroxidase; and superoxide dismutase were measured. RESULTS: Glutathione peroxidase concentration was significantly increased in dogs with IDCM, compared with control dogs. Vitamin A and superoxide dismutase concentrations were not significantly different between groups. A negative correlation was found between disease severity and plasma vitamin E concentration. Disease severity was not correlated with concentrations of other antioxidants. Medications did not significantly affect oxidant or antioxidant concentrations. CONCLUSIONS AND CLINICAL RELEVANCE: The change in glutathione peroxidase concentration and the correlation between vitamin E concentration and disease severity suggest that the oxidant-antioxidant system may play a role in development of IDCM.     

Melatonin mitigates the adverse effect of hypoxia during myocardial differentiation in mouse embryonic stem cells

BackgroundHypoxia causes oxidative stress and affects cardiovascular function and the programming of cardiovascular disease. Melatonin promotes antioxidant enzymes such as superoxide dismutase, glutathione reductase, glutathione peroxidase, and catalase.ObjectivesThis study aims to investigate the correlation between melatonin and hypoxia induction in cardiomyocytes differentiation.MethodsMouse embryonic stem cells (mESCs) were induced to myocardial differentiation. To demonstrate the influence of melatonin under hypoxia, mESC was pretreated with melatonin and then cultured in hypoxic condition. The cardiac beating ratio of the mESC-derived cardiomyocytes, mRNA and protein expression levels were investigated.ResultsUnder hypoxic condition, the mRNA expression of cardiac-lineage markers (Brachyury, Tbx20, and cTn1) and melatonin receptor (Mtnr1a) was reduced. The mRNA expression of cTn1 and the beating ratio of mESCs increased when melatonin was treated simultaneously with hypoxia, compared to when only exposed to hypoxia. Hypoxia-inducible factor (HIF)-1α protein decreased with melatonin treatment under hypoxia, and Mtnr1a mRNA expression increased. When the cells were exposed to hypoxia with melatonin treatment, the protein expressions of phospho-extracellular signal-related kinase (p-ERK) and Bcl-2-associated X proteins (Bax) decreased, however, the levels of phospho-protein kinase B (p-Akt), phosphatidylinositol 3-kinase (PI3K), B-cell lymphoma 2 (Bcl-2) proteins, and antioxidant enzymes including Cu/Zn-SOD, Mn-SOD, and catalase were increased. Competitive melatonin receptor antagonist luzindole blocked the melatonin-induced effects.ConclusionsThis study demonstrates that hypoxia inhibits cardiomyocytes differentiation and melatonin partially mitigates the adverse effect of hypoxia in myocardial differentiation by regulating apoptosis and oxidative stress through the p-AKT and PI3K pathway.